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Background: Despite the World Health Organization certifying China malaria-free in 2021, the risk of local transmission caused by imported malaria cases remains a significant clinical and public health issue. It is necessary to present the changing trends of malaria in China and discuss the role of travel medicine services in consolidating malaria elimination., Methods: This study systematically reviewed articles and reports related to human malaria from 2013 to 2022 published in international and Chinese databases. Data on malaria (i.e. number of cases, Plasmodium spp., diagnostic method, country of acquisition, provinces with high risk of re-introduction and transmission) were collected and synthesized, then summarized using descriptive statistics., Results: Overall, 24 758 cases of malaria (>99.5% laboratory confirmed, > 99.2% imported, 0.5% fatal) were reported in China from 2013 to 2022, with a downward trend over the years (4128 cases in 2013 compared to 843 cases in 2022; χ2 trend P = 0.005). The last locally acquired case was reported in 2017. Plasmodium falciparum (65.5%) was the most common species identified, followed by P. vivax (20.9%) and P. ovale (10.0%). Two P. knowlesi cases were also identified in 2014 and 2017 in returned travellers from Malaysia and Indonesia, respectively. The most common countries for malaria acquisition were Ghana, Angola and Myanmar. Plasmodium vivax was mainly detected in returned travellers from Myanmar, while P. falciparum and P. ovale were detected in travellers from sub-Saharan Africa. Imported cases were mainly reported in Yunnan, Jiangsu, Sichuan, Guangxi, Shandong, Zhejiang and Henan provinces, where large numbers of Chinese people travel overseas for work., Conclusion: Returned travellers from malaria-endemic countries pose a significant risk of malaria re-introduction to China. Travel medicine should be strengthened to improve the capacity and accessibility of both pre- and post-travel services, including malaria prophylaxis and prompt diagnosis of illness in returned travellers., (© International Society of Travel Medicine 2024. Published by Oxford University Press.)

Background: Reactive case detection (RACD) for malaria control has been found effective in low transmission settings, but its impact and cost-effectiveness in moderate-to-high transmission settings are unknown. We conducted an economic evaluation alongside an empirical trial of a modified RACD strategy (1,7-mRCTR) in three moderate-to-high malaria transmission districts in Tanzania., Methods: The costs and cost savings associated with the intervention relative to passive case detection alone were estimated in the study sites of Kilwa, Kibiti, and Rufiji districts in Tanzania from 2019-2021. Empirical cost data were collected using household surveys. The incremental costs of the intervention were calculated from under a societal perspective. Costs are reported in 2022 US dollars. Trial data and malaria registers from health facilities were used to calculate the number of malaria cases detected. We simulated unobserved and distal health effects of the intervention to assess cost-effectiveness in terms of incremental cost-effectiveness ratios (ICERs). Propagated uncertainty was assessed via second-order Monte Carlo simulation, including bootstrapping of empirical data distributions. Incremental costs per disability-adjusted life year (DALY) averted were compared to a willingness-to-pay threshold based on estimated opportunity costs of healthcare spending in Tanzania., Results: The programmatic cost of the 1,7-mRCTR intervention was 5327 United States Dollars (USD) per 1000 population. The combination of reactive and passive case detection in the intervention arm resulted in an additional 445 malaria cases detected per 1000 compared to passive detection alone, yielding an incremental cost per additional case detected of 12.0 USD. Based on modelling results, for every percentage point decline in malaria prevalence, the intervention averted 95.2 cases and 0.04 deaths per 1000 population. On average, the 1,7-mRCTR intervention averted 19.1 DALYs per 1000 population. Compared to passive malaria detection, the ICERs for the 1,7-mRCTR intervention were 7.3 USD per case averted, 16,884 USD per death averted, and 163 USD per DALY averted., Conclusions: Our analysis demonstrates that the 1,7-mRCTR intervention appears to be cost-effective under a willingness-to-pay threshold of 417 USD per DALY averted, showing that modified RACD strategies can provide value for money in moderate-to-high transmission settings., Competing Interests: Declarations. Ethics approval and consent to participate: Informed consent was obtained from heads of household and household members who were 18 years of age or above. For those under 18 years of age, informed assent was obtained from parents or guardians. Institution ethical approval was obtained from the Ifakara Health Institute Institutional Review Board (IHI/IRB/EXT/No: 18–2020) and the National Institute of Medical Research (NIMR/HQ/R.8a/Vol. IX/3634). Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Transfusion-transmitted malaria (TTM) is rare in non-endemic areas (non-EAs) but can potentially be fatal. This review analyzes the laboratory results of donors causing TTM in non-EAs, to assess the detectability of their Plasmodium infection by molecular or antibody tests., Study Design and Methods: TTM cases in the United States, Canada, and Europe since 2010 were identified through a literature review. Authors and laboratories were contacted for missing details about sample types and laboratory methods. Results of Plasmodium polymerase chain reaction (PCR) and antibody tests were summarized., Results: Twelve cases of TTM and one bone marrow transplant transmission were identified. Of the 13 source donors, 12 were tested by PCR, 10 were positive on at least one sample; the 2 negative donors were tested only on retained segments of blood refrigerated for several weeks. All donors were PCR positive on a fresh sample except one who was positive on a retained but not a fresh sample. These PCRs targeted Plasmodium DNA with sensitivities in the range of 1000-10,000 parasites/mL. Antibody EIA was positive in only three of seven donors tested., Discussion: This review found that antibody EIAs failed to detect four of the seven TTM donors tested. DNA-based PCRs were able to detect Plasmodium infection in all donors tested except for two tested only on samples likely to have deteriorated from prolonged storage. Recently developed ribosomal RNA-based molecular donor screening assays are approximately 1000 fold more sensitive than these DNA-based PCRs, holding promise as a potential method to further reduce TTM., (© 2024 The Author(s). Transfusion published by Wiley Periodicals LLC on behalf of AABB.)

Malaria is an infectious disease caused by several species of the genus Plasmodium. It is usually transmitted by female Anopheles mosquitoes. Other routes of transmission include mother-to-child transmission, shared use of needles, blood transfusion and solid organ transplantation. In non-endemic countries, malaria is often diagnosed on the basis of a history of journeys or migration from endemic areas. Transplant-transmitted malaria might represent a diagnostic challenge for clinicians. Here, we report the casual diagnosis of possible transplant-transmitted malaria in a Spanish patient with no previous visits to endemic areas. He developed symptoms one month after receiving a liver transplant from a deceased donor immigrated from Ghana. After being admitted to the Emergency Room, a complete blood count revealed an abnormal cell population which activated an 'infested red blood cells' flag (iRBC). This finding led to perform a blood smear and further tests which confirmed the diagnosis of malaria. Given that automated complete blood counts are usually performed for any patient with fever, they represent a useful tool to detect malaria in unsuspected patients. In particular, the iRBC flag implemented in Sysmex XN-Series™ hematology analyzers is a useful screening tool for malaria in clinical laboratories., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: One of the main challenge for controlling the morbidity and mortality related to malaria is the absence of reliable treatment and precise diagnostic tools in malaria-prone regions. Despite the number of diagnostic techniques used, and inconsistent results were found from previous studies. The aim of this study is to assess the effectiveness of 18S nested PCR(Polymerase Chain Reaction),microscopy, and CarestartTMpf/pan RDT(Rapid Diagnostic Tests) as malaria diagnostic methods using Microscopy and 18S nested PCR as reference techniques., Methods: A facilities and community based cross-sectional study was conducted in the villages of Batodegama kebele and Adama malaria control center from December 2016 to February 2017. A fingerprick blood sample was collected from each participant for the CarestartTM pf/pan RDT test, microscopy, and dry blood spot (DBS) analysis and 18S nested PCR assay. Data were analyzed using STATA11 to calculate the sensitivity, specificity, predictive values, and accuracy., Results: Among the 128 asymptomatic patients, 26 (20.3%) were positive for malaria nested 18S PCR, 5 (3.9%) through RDT, and 8 (6.3%) through microscopy. From 202 symptomatic Patients, 55 (27.23%), 26 (12.87%), and 28 (13.86%) individuals were positive for 18S nested PCR, RDT, and microscopy, respectively. Comparative analysis of 18S nested PCR, microscopy, and CarestartTMpf/pan RDT have similar sensitivities of 50% and specificities of 100%.for diagnosing symptomatic malaria infections. Furthermore, species level analysis showed that 18S nested PCR has a higher detection rate of 26 (20.31%) than microscopy 8 (6.25%) and CareStartTM pf/pan RDT 5 (4.1%). The CarestartTMpf/pan RDT had a sensitivity of 15.4% and specificity of 98% in detecting asymptomatic malaria infections. However, the two methods have comparable sensitivities., Conclusions: The 18S nested PCR assay showed superior performance in all circumstances. On the other hand, the overall performance of CareStart TM pf/pan RDTs was found to be comparable to that of microscopy. This finding highlights the value of parallel application of CareStart TM pf/pan RDTs and microscopy in a resource-limited setup., Competing Interests: Declarations. Ethics approval and consent to participate: Prior to the commencement of the study, ethical approval (project code 100/2017) was obtained from the Adis Abeba University College of Medicine and Health Sciences Ethical Review Committee. Permission letters were also obtained from the East Shoa Zone Health Department, Woreda Health Offices, and Oromia Regional Health Bureau. Written informed consent was obtained from all study participants who were 16 years of age or older. The purpose of the study was clearly communicated to both the participants and their parents/guardians. For children written informed consent was provided to their parents or legal guardians. Children infected with malaria were treated in accordance with national protocols and the standards declaration of Helsink. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Introduction: Contracting HIV, syphilis, hepatitis B virus (HBV), and malaria during pregnancy significantly affects the health of the woman, the pregnancy, and the unborn child. The World Health Organization (WHO) recommends testing pregnant women for these infections to achieve triple elimination of mother-to-child transmissions. However, this goal has not been fully realized in low- to medium-income countries, primarily due to segmented testing practices. This study aimed to investigate the effect of introducing a four-tests panel on the quality of antenatal care (ANC) among pregnant women attending selected Primary Health Care facilities in Kenya., Methods: Using a multi-design approach, we analyzed ANC medical records from 577 pregnant women attending eight facilities across four different counties. Blood from the women fingerpick was tested for HIV, Syphilis Hepatitis B Virus and Malaria using the four-tests panel and the results compared to those in the medical records., Results: Out of 577 ANC women, only 8.3% had test results for all four infections available. The majority of the mothers had been tested for syphilis (93.7%), HIV (78.5%), and malaria (62.6%), only 19.5% had been tested for HBV. Testing the women using the 4-tests panel yielded positivity rates of 6.9% for HIV, 0.9% for syphilis, 1.9% for malaria, and 1.1% for HBV. Among those without previous test results, the positivity rate was 2.8% for syphilis, 13.8% for HIV (with 10.6% testing positive for recent p24 infections, F = 24.876, p < 0.001), 2.3% for malaria, and 4.5% for HBV, with 83.3% of these individuals having no prior test results. The mean positivity rate of those tested using the 4-tests panel compared to segmented single tests was significantly different. The panel was cost-effective and user-friendly for healthcare workers, and in facilities facing staff shortages, it reduced turnaround time and workloads by half. The use of the panel also improved the profiling of ANC mothers and enhanced data management for the four infections by 91.7%., Conclusion: Adopting the 4-tests panel has the potential to improve test result outputs, enhance the quality-of-service delivery, and contribute significantly to the achievement of triple elimination goals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Ochwoto, Matiang’i, Machuki Onchieku, Ndoria, Matoke, Otinga, Ngondo, Zablon, Buyl, Mathebula and Matoke-Muhia.)

Objectives: Surveillance is the backbone for the control of malaria and its elimination. In the state of Gujarat, situated in the western region of India, some of the districts reported a high annual blood examination rate (ABER) for malaria. Therefore, a study was conducted to identify the underlying reasons for the increase in the ABER for malaria., Methods: Planned investigations were carried out in three of the state districts, scrutinizing records of malaria forms and other epidemiological data collected during health worker surveillance, assessment of laboratory services, and rapid fever surveys., Results: The rate of fever ranged from 8 to 57% in the primary health centers that were surveyed. Analysis of epidemiological data revealed that malaria parasite positivity was more from passive than active surveillance. Increased ABER was accounted for by multiple factors, including blood slides collected during the mass survey and contact smears, which were included in the ABER and not mentioned separately. Blood slides prepared for the migrant population were included in the ABER, but the migrant population was not counted while calculating the ABER., Conclusion: The ABER in villages surveyed varied from 1.6 to 78%, which is mainly due to indiscriminate preparation of blood slides, i.e., without fever symptoms. Addressing the key gaps identified in data recording may aid in channeling the limited resources efficiently, thereby progressing toward malaria elimination. Adequate surveillance activities, along with systematic data recording, will enable timely, informed decision-making for the effective allocation of resources, ultimately supporting malaria elimination efforts in the state., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Kaur, Baharia and Dattani.)

Background: Malaria is a major cause of ill health and death in children in Africa. The disease also imposes a severe social and economic burden on households and health systems and is strongly associated with poverty. This study summarizes the most up-to-date cost of treating severe malaria in children in Africa., Methods: A systematic search was conducted in PubMed, Embase, Cinahl, and Web of Science databases. Google and Google Scholar were searched for grey literature followed by scanning of the reference lists of the previous systematic reviews. The search was limited to children < 12 years, malaria-endemic countries in Africa, and the English language. All costs were adjusted to the year 2023., Results: 19 studies conducted in 12 countries were identified: 14 reported provider costs, and 11 household costs. Out of the 19 studies found, 11 were published before 2018 while 11 reported data that are currently more than ten years old. Studies varied methodologically and in the scope of resources included to estimate the cost. The provider costs ranged from USD 27 in Uganda to USD 165 per patient in Kenya (median value USD 90), while household costs ranged from USD 13 in Kenya to USD 245 per patient in Gabon (median value USD 50). All identified household malaria treatment costs except one represented catastrophic health expenditure, making out more than 10% of the monthly Gross National Income per capita in the respective countries., Conclusion: Evidence on the cost of treating severe malaria in children in Africa is scarce. However, the few existing studies show that severe malaria in children imposes a significant economic burden on the providers and households. More studies are needed, particularly in high-burden high-impact countries, to inform resource allocation decisions., (© 2024. The Author(s).)

Background: Malaria is a global disease caused by the transmission of the malaria parasite through the bites of infected Anopheles mosquitoes. There are 4 kinds of common malaria parasites, among which oval malaria is mainly prevalent in tropical West Africa, with a narrow distribution range, mainly imported cases in China, which is relatively rare. Because the morphology, attack cycle, recrudescence and relapse are similar to those of Plasmodium vivax and, therefore, is easily missed and misdiagnosed., Methods: Malaria parasite-specific antigens were detected in whole blood samples using rapid diagnostic tests (RDT). Peripheral blood was used to prepare thick and thin smears, which were then stained with Giemsa for the identification of malaria parasites and different parasite morphologies under an oil microscope. Plasmodium species were identified using fluorescence quantitative PCR., Results: The patient's RDT revealed two red response lines (only T2 positive), indicating a single or mixed infection of three types of malaria (Plasmodium vivax, Plasmodium malarie, Plasmodium ovale) excluding Plasmodium falciparum. Thick smears exhibited various stages of Plasmodium such as hypertrophic rings and gametocytes, while thin films displayed Plasmodium trophozoites, gametocytes, etc., resembling oval malaria. The patients were diagnosed with Plasmodium ovum infection through fluorescence quantitative PCR, and targeted treatment was administered., Conclusions: The incidence of Plasmodium ovum infection is low, and there is a risk of overlooking or misdiagnosing the infection in laboratory tests. It is essential for laboratory staff to enhance their morphological recognition skills for Plasmodium and to integrate blood routine analysis, RDT, and PCR results in order to facilitate early diagnosis and prompt treatment.

Malaria remains a major global health problem. Transmission occurs in 84 countries across five continents, with almost 250 million cases and over 600,000 deaths each year. Primary and secondary care clinicians in the UK need to be alert to the prospect of malaria presenting in returning travellers. They must be aware of the signs of severe malaria, the need for prompt diagnosis and treatment, and the importance of seeking specialist advice. With emerging resistance, climate change and the roll-out of the first malaria vaccines, the landscape of malaria is changing. Here we discuss the past, present and future of malaria., Competing Interests: Declaration of competing interest Angela M Minassian has an immediate family member who is an inventor on patent applications relating to RH5 malaria vaccines. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Recent studies by Girgis et al. and de Cesare et al. promise to advance malaria genomic surveillance using inexpensive and portable long-read amplicon-sequencing technologies. These technologies allow rapid characterization of drug-resistance markers, antigenic diversity, and diagnostic target loci from dried blood spots, providing new tools for surveillance in endemic regions and informing interventions to combat malaria more effectively., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Purpose: Malaria keeps on being a serious global health threat, especially in many tropical countries, where it is endemic. Also in non-endemic countries, like Spain, malaria is an issue that requires attention due to the presence of imported cases., Methods: This is a retrospective study, including all patients diagnosed with malaria at Severo Ochoa University Hospital from 2006 to 2022, being classified according to: (I) their type of stay in an endemic area as visiting friends and relatives (VFR), migrants of recent arrival (MRA), or tourism and business (T&B), and (II) the mode of presentation as microscopic (MM) or submicroscopic (SMM) malaria., Results: In this study, 132 patients (23.7% of all suspected) were diagnosed with malaria. The PCR was the most sensitive technique (99.2%), followed by antigen detection (78.8%) and microscopy (75%), with Plasmodium falciparum being the predominant species (94.7%). VFR was the largest group infected with malaria (69.7%), mostly symptomatic (98.2%) and presenting MM (90.2%). Instead, MRA patients (25%) presented milder (47.4%) or no symptoms (31.6%) and higher cases of SMM (42.4%). Coinfection with another imported pathogen was present in 19 patients (14.4%), being MRA more frequently coinfected (30.3%) CONCLUSION: This study shows the need for establishing systems for VFRs to attend pre-travel consultations to reduce malaria imported risk. In the case of MRA, screening for imported diseases should be conducted upon their arrival. Finally, we highlight two cases of co-infection with imported viruses, showing that presence of symptoms resembling malaria from another imported pathogen does not exclude malaria., (© 2024. The Author(s).)

The apical membrane antigen-1 (AMA-1) is a crucial target for malaria management and prevention strategies. While the immunogenicity of AMA-1 has been extensively studied for Plasmodium falciparum and Plasmodium vivax, there is a notable scarcity of information for Plasmodium malariae. In this study, recombinant PmAMA-1 was expressed in Escherichia coli, and its integrity was confirmed via western blotting and indirect immunofluorescence assays. Immunization of BALB/c mice with rPmAMA-1 emulsified in Freund's adjuvant resulted in significantly elevated specific IgG antibodies, predominantly IgG1. The immune response exhibited Th1, Th2, and Th17 phenotypes, with a notable Th1 bias. Antisera from immunized mice effectively recognized native PmAMA-1 on P. malariae. These results suggest that PmAMA-1 is a promising target for both vaccine development and diagnostic applications for P. malariae infections, offering dual preventive and diagnostic benefits in malaria control., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

Background: Resource-limited regions face a greater burden of infectious diseases due to limited access to molecular tests, complicating timely diagnosis and management. Current molecular point-of-care tests (POCTs) either come with high costs or lack adequate sensitivity and specificity. To facilitate better prevention and control of infectious diseases in underserved areas, we seek to address the need for molecular POCTs that better align with the World Health Organization (WHO)'s ASSURED criteria-Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free, and Deliverable to end users., Methods: A novel molecular POCT, Pasteur Pipette-assisted isothermal probe amplification (pp-IPA), was developed for malaria detection. Without any microfluidics, this method captures Plasmodium 18S rRNA in a modified Pasteur pipette using tailed genus-specific probes. After washing, the bound tailed probes are ligated to form a template for subsequent novel isothermal probe amplification using a pair of generic primers, bypassing nucleic acid extraction and reverse transcription. The method was assessed using cultured Plasmodium and compared with real-time quantitative reverse transcription PCR (RT-qPCR) or reverse transcription loop-mediated isothermal amplification (RT-LAMP) in clinical blood samples., Results: The entire assay is completed in 60-80 min with minimal hands-on time, using only a Pasteur pipette and a water bath. The pp-IPA's analytical sensitivity is 1.28 × 10 -4  parasites/μl, with 100% specificity against various blood-borne pathogens causing malaria-like symptoms. Additionally, pp-IPA needs only liquid-transfer skill for operation and the cost is around USD 0.25 per test, making it at least 300 times lower than mainstream POCT platforms., Conclusions: Designed to improve the accessibility of molecular detection in resource-limited settings, pp-IPA's simplicity, affordability, high sensitivity/specificity, and minimal equipment requirements make it a promising point-of-care pathogen identification tool in resource-constrained regions., (© 2024. The Author(s).)

Background: Reports on the impact of COVID-19 pandemic on the quality of malaria care and burden in sub Saharan Africa have provided a mixed picture to date. The impact of the 2nd (Delta) and 3rd (Omicron) COVID-19 waves on outpatient malaria indicators and case management practices was assessed at three public health facilities with varying malaria transmission intensities in Uganda., Methods: Individual level data from all patients presenting to the out-patient departments (OPD) of the three facilities (Kasambya, Walukuba and Lumino) between January 2019 and February 2022 were included in the analysis. Outcomes of interest included total number of outpatient (OPD) visits, proportion of patients suspected to have malaria, proportion of suspected malaria cases tested with a malaria diagnostic test, test positivity rates (TPR) and proportion of malaria cases prescribed artemether-lumefantrine (AL). Using the pre-COVID-19 trends between January 2019 and February 2020, interrupted time series analysis was used to predict the expected trends for these study outcomes during the 2nd wave (May 2021-August 2021) and 3rd wave (November 2021-February 2022). The observed trends of the study outcomes were compared with the expected trends., Results: There were no significant differences between the observed versus expected overall outpatient visits in the 2nd wave, however, a significant decline in OPD attendance was observed during the 3rd wave (15,101 vs 31,154; incidence rate ratio (IRR) = 0.48 [0.41-0.56]). No significant differences in the overall observed versus expected proportions of suspected malaria cases and test positivity rates in both COVID waves. However, a significant decrease in the overall proportion of suspected malaria cases tested with a malaria diagnostic test was observed during the 3rd wave (99.86% vs 99.99%; relative percent ratio [RPR] = 0.99 [0.99-0.99]). Finally, a significant decline in the overall proportion of malaria cases prescribed AL was observed during the 2nd wave (94.99% vs 99.85%; RPR = 0.95 [0.92-0.98]) but not the 3rd wave., Conclusion: Significant declines in OPD attendance and suspected malaria cases tested with malaria diagnostic test were observed during the 3rd COVID-19 wave, while AL prescription significantly reduced during the 2nd COVID-19 wave. These findings add to the body of knowledge highlighting the adverse impact of COVID-19 pandemic on the malaria which could explain the increase in the malaria burden observed during this period., (© 2024. The Author(s).)

Importance: A prompt malaria diagnosis is crucial for the management of children with febrile illness in sub-Saharan African countries, where malaria remains a leading cause of mortality among children younger than 5 years of age. The development and distribution of point-of-care rapid diagnostic tests (RDTs) for malaria has transformed practice, but limited systematic evidence exists on how malaria RDTs have affected the management of febrile illness and mortality for children younger than 5 years of age across sub-Saharan Africa countries., Objective: To evaluate the association between the distribution of malaria RDTs and the management of febrile illness and mortality among children younger than 5 years of age in sub-Saharan African countries., Design, Setting, and Participants: This quasi-experimental study used a novel dataset linking malaria RDT distribution to 165 nationally representative household surveys across 35 sub-Saharan African countries with mortality data. The sample comprised approximately 3.9 million child-year observations and approximately 260 000 febrile illness episodes in children younger than 5 years of age between 2000 and 2019., Main Outcomes and Measures: Fixed-effects linear probability models were used to analyze the association between variation in malaria RDTs distributed per child younger than 5 years of age (by country per year) and blood testing, antimalarial drug use, antibiotic use, use of symptomatic treatments, and mortality rates. Variation in the effects of testing and treatment was also assessed across the sub-Saharan African countries that had varying prevalence of malaria., Results: The mortality sample included 1 317 866 children and the fever sample included 256 292 children. The mean age of the children with febrile illness was 2.4 years (SD, 1.3 years) and 49% were female. Each additional malaria RDT distributed per child younger than 5 years of age was associated with an increase of 3.5 percentage points (95% CI, 3.2-3.8 percentage points) in blood testing, an increase of 1.5 percentage points (95% CI, 1.2-1.8 percentage points) in the use of antimalarial drugs, an increase of 0.4 percentage points (95% CI, 0.1-0.6 percentage points) in antibiotic use, and a decrease of 0.4 percentage points (95% CI, 0.1-0.8 percentage points) in the use of treatments for symptoms. Each additional malaria RDT distributed per child younger than 5 years of age was associated with a reduction in child mortality of 0.34 deaths per 1000 child-years (95% CI, 0.15-0.52 deaths per 1000 child-years). The effects of malaria RDT distribution on medication use and child mortality varied across prevalence settings (low vs high) for malaria; there were survival improvements only in areas that had a high prevalence of malaria., Conclusions and Relevance: Increasing distribution of malaria RDTs was associated with increased blood testing, increased use of antimalarial drugs, and modestly improved survival in children younger than 5 years of age in sub-Saharan African countries. However, malaria RDTs were associated with increases in the rates of antibiotic use that were already high, suggesting that more comprehensive approaches to case management of febrile illness are needed.

The identification of malaria infection using microscope images of blood smears is considered as a 'gold standard'. The diagnosis of malaria needs expert microscopists which are scarce in remote areas where malaria is endemic. Therefore, it is desirable to automate the repetitive task of pathogen detection in the blood samples received as microscope images. This study provides an easy to use and deploy method for implementing a malaria pathogen detection software- the Intelligent Suite. The Intelligent Suite features a graphical user interface (GUI) implemented using 'cvui' library to interact with the OpenVINO's inference engine for model optimisation and deployment across several inference devices. The intelligent Suite uses a custom YOLO-mp-3l model trained on Darknet framework for detection of malaria pathogen in thick smear microscope images. Moreover, the Intelligent Suite provides user interface for inference device/mode selection, alter model parameters, and generate detection reports along with the model performance metrics. The Intelligent Suite was executed on a CPU computer with model inference running on a plug-and-play Neural Compute Stick (NCS2) and performance reported., (© 2024. The Author(s).)

Plasmodium spp. infections and cases of malaria are a long-standing public health problem for children living in middle- and low-income countries. Dengue virus causes an emerging under-recognized disease burden. A cross sectional study was conducted between March 2020 and December 2021 to determine the status of malaria and dengue fever, and the associated factors in children living in Mwanza, Tanzania. Clinical features were recorded; blood samples were analyzed using dengue NS1 rapid diagnostics test (NS1-RDT), malaria rapid diagnostic test (MRDT) and PCR and microscopy for malaria parasites. Descriptive analysis was based on infection status; odds ratio and confidence interval were used to determine the factors associated with dengue fever and malaria. The prevalence of malaria in the 436 children included in the final analysis was 15.6%, 8.5%, and 12.1% as determined by MRDT, blood smear examination and PCR, respectively. The prevalence of dengue fever determined by the NS1-RDT was 7.8%. Body rash, muscle and joint/bone pain were associated with a positive rapid dengue test result. Retro-orbital pain characterized Plasmodium spp. and dengue virus co-infections. Clinical signs and symptoms could not readily differentiate between malaria and dengue fever patients or patients co-infected with both causative agents underscoring the urgent need for the accurate laboratory diagnostics. Additional large-scale studies are required to assess the epidemiological burden of acute febrile illness in developing countries and to produce data that will guide empirical treatment., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Kayange et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: COVID-19 disruptions including lockdowns and prioritization of COVID-19 control programs in Africa in 2020-2022 contributed to reductions in malaria control activities including malaria diagnosis, treatment and resistance monitoring. This study investigated the malaria burden and distribution on the background of active transmission of SARS-CoV-2 in Southern Ghana; utilizing community health education and medical screening (CHEMS) approach to determine epidemiological overlaps in COVID-19 and malaria., Methods: Between October-December 2022, prospective cross-sectional surveys, with CHEMS were conducted in Greater Accra and Central regions, where 994 participants enrolled either at a hospital or community setting provided demographic and clinical data including history of clinical malaria infection and antimalarial treatment in the past 2 weeks. Of this study population, 953 provided nasal/throat swabs for COVID-19 RT-PCR testing, with a subset of 136 participants also providing finger-prick blood for malaria RDT testing., Results: The study population comprised of 73.6% adults, with 54.6% COVID-19 vaccination rate. Overall, 18.1% of participants had a history of clinical malaria, which was associated (adjusted odds ratio > 1.50, p -value ≤0.022) with COVID-19 symptoms and positivity, study area and hospital setting, suggestive of overlaps in the epidemiological risk for malaria. On a background of widespread SARS-CoV-2 infections (12-37%), malaria parasitaemia was detected in 6%, with 2% being co-infections with SARS-CoV-2. Among the malaria positives, 9.5% had a history of antimalarial treatment, which suggested that their infections were recrudescent parasitaemia., Conclusion: The epidemiological and clinical overlap between malaria and COVID-19 within the hospital and community settings underscores the need for accurate case diagnosis to inform effective clinical treatments. Innovative surveillance programs, with community engagement are needed to maximize control interventions including treatment of asymptomatic malaria infections., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Amegatcher, Acquah, Tetteh, Obeng, Debrah, Quist, Acquah-Jackson, Boateng, Twieku, Armoo, Awandare, Mosi and Narh.)

BackgroundIn European France, the bulk of malaria cases are travel-related, and only locally acquired cases are notifiable to assess any risk of re-emergence.AimsWe aimed to contribute to assessing the health impact of locally acquired malaria and the potential of malaria re-emergence in European France by documenting modes of transmission of locally acquired malaria, the Plasmodium species involved and their incidence trends.MethodsWe retrospectively analysed surveillance and case investigation data on locally acquired malaria from 1995 to 2022. We classified cases by most likely mode of transmission using a classification derived from the European Centre for Disease Prevention and Control. A descriptive analysis was conducted to identify spatial and temporal patterns of cases.ResultsFrom 1995 to 2022, European France reported 117 locally acquired malaria cases, mostly due to Plasmodium falciparum (88%) and reported in Île-de-France (54%), Paris Region. Cases were classified as Odyssean malaria (n = 51), induced malaria (n = 36), cryptic malaria (n = 27) and introduced malaria (n = 3). Among the 117 patients, 102 (93%) were hospitalised, 24 (22%) had severe malaria and seven (7%) died.ConclusionLocally acquired malaria remains infrequent in European France, with four reported cases per year since 1995. However, with the recent increasing trend in Odyssean malaria and climate change, the risk of re-emergence in non-endemic countries should be monitored, particularly in areas with autochthonous competent vectors. The vital risk of delayed diagnosis should make physicians consider locally acquired malaria in all patients with unexplained fever, especially when thrombocytopenia is present, even without travel history.

The Intergovernmental Panel on Climate Change has reported that the prevalence of vector-borne diseases has increased in recent decades and that the prevalence of malaria, Lyme disease, dengue, and, in particular, West Nile virus infection are expected to increase further if control measures are not strengthened. (1)(2) This review article summarizes the epidemiology, various clinical manifestations, and management strategies of these vector-borne diseases with increasing prevalence both in the United States and worldwide., (© American Academy of Pediatrics, 2024. All rights reserved.)

Malaria and babesiosis are global health threats affecting humans, wildlife, and domestic animals, particularly in Africa, the Americas, and Europe. Malaria can lead to severe outcomes, while babesiosis usually resembles a mild illness but can be severe and fatal in individuals with weakened immune systems. Swift, accurate detection of these parasites is crucial for treatment and control. We evaluated a real-time PCR assay for diagnosing five Plasmodium and three Babesia species from blood samples, assessing its sensitivity, specificity, and analytical performance by analyzing 46 malaria-positive and 32 Babesia spp-positive samples diagnosed through microscopy. The limit of detection for Plasmodium species ranged from 30 to 0.0003 copies/µL. For mixed infections, it was 0.3 copies/µL for P. falciparum/P. vivax and 3 copies/µL for P. malariae/P. knowlesi. Babesia species had a detection limit of 0.2 copies/µL. No cross-reactivity was observed among 64 DNA samples from various microorganisms. The assay showed good sensitivity, detecting Plasmodium and Babesia species with 100 % accuracy overall, except for P. falciparum (97.7 %) and B. microti (12.5 %). The low sensitivity of detecting B. microti was attributed to limitations in microscopy for species identification. This technique heavily relies on the proficiency of the examiner, as species within the genus cannot be distinguished under a microscope. Additionally, Babesia can be confused with the early trophozoite stage (ring forms) of Plasmodium parasites. The findings support multiplex qPCR's diagnostic superiority over the gold standard, despite higher costs. It offers enhanced sensitivity, specificity, and detects mixed infections, crucial for effective monitoring and diagnosis of malaria and babesiosis in endemic regions with significant public health challenges., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Purpose of Review: Malaria threatens pregnant women and their babies, particularly in Africa., Recent Findings: This century, the number of women at risk of malaria in pregnancy has decreased globally, apart from in Africa, where it has increased. Low and sub microscopic infections are increasingly documented but remain hard to diagnose with current point-of-care tests, and their contribution to morbidity and transmission are unclear. Artemether-lumefantrine has been endorsed for treatment in first trimester, but many women attend antenatal clinics later in pregnancy, and reaching high-risk young, first-time mothers is particularly difficult. Small-for-gestational-age babies frequently result from malaria, which affects the placenta's development and its functions such as nutrient transport. Resistance to continues to increase to sulphadoxine-pyrimethamine, the mainstay of intermittent preventive treatment in pregnancy. The alternative, dihydroartemisinin-piperaquine controls malaria better, but does not improve pregnancy outcomes, suggesting that sulphadoxine-pyrimethamine may have nonmalarial effects including improving gut function or reducing dangerous inflammation. Understanding of how the malaria parasite uses the VAR2CSA protein to bind to its placental receptor is increasing, informing the search for a vaccine to prevent pregnancy malaria., Summary: Progress in several areas increases optimism that improved prevention and control of malaria in pregnancy is possible, but obstacles remain., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Malaria is a deadly disease of significant concern for the international community. It is an infectious disease caused by a Plasmodium spp. parasite and transmitted by the bite of an infected female Anopheles mosquito. The parasite multiplies in the liver and then destroys the person's red blood cells until it reaches the severe stage, leading to death. The most used tools for diagnosing this disease are the microscope and the rapid diagnostic test (RDT), which have limitations preventing control of the disease. Computer vision technologies present alternatives by providing the means for early detection of this disease before it reaches the severe stage, facilitating treatment and saving patients. In this article, we suggest deep learning methods for earlier and more accurate detection of malaria parasites with high generalization capabilities using microscopic images of blood smears from many heterogeneous patients. These techniques are based on an image preprocessing method that mitigates some of the challenges associated with the variety of red cell characteristics due to patient diversity and other artifacts present in the data. For the study, we collected 65,970 microscopic images from 876 different patients to form a dataset of 33,007 images with a variety that enables us to create models with a high level of generalization. Three types of convolutional neural networks were used, namely Convolutional Neural Network (CNN), DenseNet, and LeNet-5, and the highest classification accuracy on the test data was 97.50% found with the DenseNet model., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Raman spectroscopy was used to study erythrocytes collected from patients diagnosed with malaria at the University Hospital in Kraków and from healthy volunteers. A laser line with a wavelength of 442 nm was used to induce the Raman resonance of haem, while a laser with a wavelength of 785 nm was used for the normal Raman effect. The results were analysed using Principal Component Analysis. For the 442 nm laser line, analysis of the entire spectral range (3200 cm -1 to 300 cm -1 ) showed satisfactory separation of Raman spectra for healthy cells from infected cells, which was significantly improved in the 1500 cm -1 -1200 cm -1 spectral range. For the 785 nm laser line, some separation was observed in each range studied, but the best results were achieved over the full spectral range. Plasmodium-derived nucleic acids and phosphodiester vibrations were observed at excitation lines of 442 nm and 785 nm, respectively., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Malaria is the parasitic disease with the highest morbimortality worldwide. The World Health Organization (WHO) estimates that there were approximately 249 million cases in 2022, of which 3.4% were in Angola. Diagnosis is based on parasite identification by microscopy examination, antigen detection, and/or molecular tests, such as polymerase chain reaction (PCR). This study aimed to evaluate the usefulness of real-time PCR as a diagnostic method for malaria in an endemic area (Cubal, Angola)., Methods: A cross-sectional study was carried out at the Hospital Nossa Senhora da Paz in Cubal, Angola, including 200 patients who consulted for febrile syndrome between May and July 2022. From each patient, a capillary blood sample was obtained by finger prick for malaria field diagnosis [microscopy and rapid diagnostic test (RDT)] and venous blood sample for real-time PCR performed at the Hospital Universitario Vall d'Hebron in Barcelona, Spain. Any participant with a positive result from at least one of these three methods was diagnosed with malaria., Results: Of the 200 participants included, 54% were female and the median age was 7 years. Malaria was diagnosed by at least one of the three techniques (microscopy, RDT, and/or real-time PCR) in 58% of the participants, with RDT having the highest percentage of positivity (49%), followed by real-time PCR (39.5%) and microscopy (33.5%). Of the 61 discordant samples, 4 were only positive by microscopy, 13 by real-time PCR, and 26 by RDT. Plasmodium falciparum was the most frequent species detected (90.63%), followed by P. malariae (17.19%) and P. ovale (9.38%). Coinfections were detected in ten participants (15.63%): six (60%) were caused by P. falciparum and P. malariae, three (30%) by P. falciparum and P. ovale, and one (10%) triple infection with these three species. In addition, it was observed that P. falciparum and P. malariae coinfection significantly increased the parasite density of the latter., Conclusions: RDT was the technique with the highest positivity rate, followed by real-time PCR and microscopy. The results of the real-time PCR may have been underestimated due to suboptimal storage conditions during the transportation of the DNA eluates. However, real-time PCR techniques have an important role in the surveillance of circulating Plasmodium species, given the epidemiological importance of the increase in non-falciparum species in the country, and can provide an estimate of the intensity of infection., (© 2024. The Author(s).)

Objective: To perform critical methodological assessments on designs, outcomes, quality and implementation limitations of studies evaluating the impact of malaria rapid diagnostic tests (mRDTs) on patient-important outcomes in sub-Saharan Africa., Design: A systematic review of study methods., Data Sources: MEDLINE, EMBASE, Cochrane Library, African Index Medicus and clinical trial registries were searched up to May 2022., Eligibility Criteria: Primary quantitative studies that compared mRDTs to alternative diagnostic tests for malaria on patient-important outcomes within sub-Sahara Africa., Data Extraction and Synthesis: Studies were sought by an information specialist and two independent reviewers screened for eligible records and extracted data using a predesigned form using Covidence. Methodological quality was assessed using the National Institutes of Health tools. Descriptive statistics and thematic analysis guided by the Supporting the Use of Research Evidence framework were used for analysis. Findings were presented narratively, graphically and by quality ratings., Results: Our search yielded 4717 studies, of which we included 24 quantitative studies; (15, 62.5%) experimental, (5, 20.8%) quasi-experimental and (4, 16.7%) observational studies. Most studies (17, 70.8%) were conducted within government-owned facilities. Of the 24 included studies, (21, 87.5%) measured the therapeutic impact of mRDTs. Prescription patterns were the most reported outcome (20, 83.3%). Only (13, 54.2%) of all studies reported statistically significant findings, in which (11, 45.8%) demonstrated mRDTs' potential to reduce over-prescription of antimalarials. Most studies (17, 70.8%) were of good methodological quality; however, reporting sample size justification needs improvement. Implementation limitations reported were mostly about health system constraints, the unacceptability of the test by the patients and low trust among health providers., Conclusion: Impact evaluations of mRDTs in sub-Saharan Africa are mostly randomised trials measuring mRDTs' effect on therapeutic outcomes in real-life settings. Though their methodological quality remains good, process evaluations can be incorporated to assess how contextual concerns influence their interpretation and implementation., Prospero Registration Number: CRD42018083816., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY. Published by BMJ.)

Rodent malaria models serve as important preclinical antimalarial and vaccine testing tools. Evaluating treatment outcomes in these models often requires manually counting parasite-infected red blood cells (iRBCs), a time-consuming process, which can be inconsistent between individuals and laboratories. We have developed an easy-to-use machine learning (ML)-based software, Malaria Screener R, to expedite and standardize such studies by automating the counting of Plasmodium iRBCs in rodents. This software can process Giemsa-stained blood smear images captured by any camera-equipped microscope. It features an intuitive graphical user interface that facilitates image processing and visualization of the results. The software has been developed as a desktop application that processes images on standard Windows and MacOS computers. A previous ML model created by the authors designed to count Plasmodium falciparum-infected human RBCs did not perform well counting Plasmodium-infected mouse RBCs. We leveraged that model by loading the pretrained weights and training the algorithm with newly collected data to target Plasmodium yoelii- and Plasmodium berghei-infected mouse RBCs. This new model reliably measured both P. yoelii and P. berghei parasitemia (R2 = 0.9916). Additional rounds of training data to incorporate variances due to length of Giemsa staining and type of microscopes, etc., have produced a generalizable model, meeting WHO competency level 1 for the subcategory of parasite counting using independent microscopes. Reliable, automated analyses of blood-stage parasitemia will facilitate rapid and consistent evaluation of novel vaccines and antimalarials across laboratories in an easily accessible in vivo malaria model.

Background: The parasite species Plasmodium ovalecurtisi (P. ovalecurtisi) and Plasmodium ovalewallikeri (P. ovalewallikeri), formerly known as Plasmodium ovale, are endemic across multiple African countries. These species are thought to differ in clinical symptomatology and latency, but only a small number of existing diagnostic assays can detect and distinguish them. In this study, we sought to develop new assays for the detection and differentiation of P. ovalecurtisi and P. ovalewallikeri by leveraging recently published whole-genome sequences for both species., Methods: Repetitive sequence motifs were identified in available P. ovalecurtisi and P. ovalewallikeri genomes and used for assay development and validation. We evaluated the analytical sensitivity of the best-performing singleplex and duplex assays using synthetic plasmids. We then evaluated the specificity of the duplex assay using a panel of samples from Tanzania and the Democratic Republic of the Congo (DRC), and validated its performance using 55 P. ovale samples and 40 non-ovale Plasmodium samples from the DRC., Results: The best-performing P. ovalecurtisi and P. ovalewallikeri targets had 9 and 8 copies within the reference genomes, respectively. The P. ovalecurtisi assay had high sensitivity with a 95% confidence lower limit of detection (LOD) of 3.6 parasite genome equivalents/μl, while the P. ovalewallikeri assay had a 95% confidence LOD of 25.9 parasite genome equivalents/μl. A duplex assay targeting both species had 100% specificity and 95% confidence LOD of 4.2 and 41.2 parasite genome equivalents/μl for P. ovalecurtisi and P. ovalewallikeri, respectively., Conclusions: We identified promising multi-copy targets for molecular detection and differentiation of P. ovalecurtisi and P. ovalewallikeri and used them to develop real-time PCR assays. The best performing P. ovalecurtisi assay performed well in singleplex and duplex formats, while the P. ovalewallikeri assay did not reliably detect low-density infections in either format. These assays have potential use for high-throughput identification of P. ovalecurtisi, or for identification of higher density P. ovalecurtisi or P. ovalewallikeri infections that are amenable to downstream next-generation sequencing., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: JBP reports research support from Gilead Sciences, non-financial support from Abbott Laboratories, and consulting for Zymeron Corporation, all outside the scope of the manuscript. All other authors declare no competing interests., (Copyright: © 2024 He et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Malaria control depends primarily on rapid and accurate diagnosis followed by successful treatment. Light microscopy is still used as a gold standard method for the diagnosis of malaria. The Sysmex hematology analyzer is a novel method for malaria detection. Therefore, the aim of this review was to investigate the diagnostic accuracy of the Sysmex hematology analyzer for malaria diagnosis., Methods: Electronic databases like PubMed, PubMed Central, Science Direct databases, Google Scholar, and Scopus were used to find relevant articles from April to June 14, 2023. The studies' methodological quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies-2 tool. Using Review Manager 5.4.1, the estimates of sensitivity and specificity, as well as their 95% confidence intervals, were shown in forest plots. Midas software in Stata 14.0 was utilized to calculate the summary estimates of sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio. Heterogeneity was assessed by using I2 statistics. In addition, publication bias was assessed using a funnel plot and Deeks' test. Sub-group and meta- regression analysis were also performed., Results: A total of 15 studies were assessed for diagnostic accuracy. The sensitivity and specificity of Sysmex hematology analyzer for studies ranged from 46% to 100% and 81% to 100%, respectively. The summary estimate of sensitivity and specificity of Sysmex hematology analyzer were 95% (95% CI: 85%-99%) and 99% (95% CI: 97%-100%), respectively. It had excellent diagnostic accuracy. There were significant heterogeneity among the studies included in this meta-analysis. The summary estimate of sensitivity and specificity of Sysmex hematology analyzer using polymerase chain reaction as the gold standard was 97.6% (95% CI: 83.2, 99.7) and 99.4% (98.5, 99.8), respectively., Conclusion: In this review, Sysmex hematology analyzer had excellent diagnostic accuracy. Therefore, it could be used as an alternate diagnostic tool for malaria diagnosis in the hospital and health center., Trial Registration: Systematic review registration PROSPERO (2023: CRD42023427713). https://www.crd.york.ac.uk/prospero/display&#95;record.php?ID=CRD42023427713., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mulatie et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Plasmodium knowlesi, primarily a zoonotic malaria species is the most common malaria pathogen in the Southeast Asia especially in Malaysian Borneo, Malaysia. Due to morphological resemblance of P. knowlesi to other human Plasmodium, the sensitivity for microscopic detection of P. knowlesi, which is the gold standard, is compromised. Thus, efforts have been made in finding alternatives for the disease diagnosis. This study described the potential use of anti-PkTRAMP polyclonal antibodies in sandwich ELISA for P. knowlesi detection. Anti-PkTRAMP polyclonal antibodies raised from mice and rabbit were first evaluated for their binding capability towards native proteins in P. knowlesi lysates using Western blot. These mice and rabbit polyclonal antibodies were then used in the sandwich ELISA as capture and detection antibodies, respectively. P. knowlesi A1H1 culture was utilised to determine the limit of detection (LOD) of this assay. Its clinical performance was determined by testing with archived human malaria and uninfected samples. Western blot analysis affirmed the polyclonal antibodies reactivity to P. knowlesi. The LOD obtained from three replicated assays was at 0.015% parasitaemia. The assay has 76% sensitivity and 75% specificity for P. knowlesi. Its positive and negative predictive values were 76% and 75%, respectively. No cross reactivity with P. falciparum and healthy samples was observed, except for P. vivax where 10 out of 12 samples were detected. In conclusion, anti-PkTRAMP polyclonal antibodies can be useful in detecting P. knowlesi. Regardless, the full potential of anti-PkTRAMP antibodies for diagnostic purposes need to be explored further.

Background and Objectives: The risk of transfusion-transmitted malaria (TTM) infections is extremely low in Australia, and the cost-effectiveness of the current screening strategy has not been assessed. This study aims to conduct a cost-effectiveness analysis of different malaria screening strategies in blood donors as part of the risk-based decision-making framework., Materials and Methods: A decision tree model was developed to assess the cost-effectiveness of five alternative malaria screening strategies from a healthcare sector perspective. Screening strategies combining total or partial removal of malaria testing with different deferral periods were considered. The probabilities of developing severe and uncomplicated TTM were based on a literature review of cases in non-endemic areas since 2000. The health outcomes were quantified using disability-adjusted life years. The costs of non-returning donors due to deferral were also included. Deterministic and probabilistic sensitivity analyses were conducted to account for data uncertainty., Results: The residual risks for all strategies were so low that the costs, mortality and morbidity associated with TTM are almost negligible. The overall costs were predominantly influenced by the costs of non-returning blood donors. As a result, removal of malaria testing and applying a 28-day deferral for at-risk donors were the least costly and most cost-effective of all the options considered., Conclusion: The current screening strategy for malaria in blood donors in Australia is not an efficient use of healthcare resources. Partial or total removal of malaria testing would bring significant cost savings without significantly compromising blood safety., (© 2024 The Author(s). Vox Sanguinis published by John Wiley & Sons Ltd on behalf of International Society of Blood Transfusion.)

Spread of the Anopheles stephensi mosquito, an invasive malaria vector, threatens to put an additional 126 million persons per year in Africa at risk for malaria. To accelerate the early detection and rapid response to this mosquito species, confirming its presence and geographic extent is critical. However, existing molecular species assays require specialized laboratory equipment, interpretation, and sequencing confirmation. We developed and optimized a colorimetric rapid loop-mediated isothermal amplification assay for molecular An. stephensi species identification. The assay requires only a heat source and reagents and can be used with or without DNA extraction, resulting in positive color change in 30-35 minutes. We validated the assay against existing PCR techniques and found 100% specificity and analytical sensitivity down to 0.0003 ng of genomic DNA. The assay can successfully amplify single mosquito legs. Initial testing on samples from Marsabit, Kenya, illustrate its potential as an early vector detection and malaria mitigation tool.

The African continent carries the greatest malaria burden in the world. Falciparum malaria especially has long been the leading cause of death in Africa. Climate, economic factors, geographical location, human intervention and unstable security are factors influencing malaria transmission. Due to repeated infections and early interventions, the proportion of clinically atypical malaria or asymptomatic plasmodium carriers has increased significantly, which easily lead to misdiagnosis and missed diagnosis. African countries have made certain progress in malaria control and elimination, including rapid diagnosis of malaria, promotion of mosquito nets and insecticides, intermittent prophylactic treatment in high-risk groups, artemisinin based combination therapies, and the development of vaccines. Between 2000 and 2022, there has been a 40% decrease in malaria incidence and a 60% reduction in mortality rate in the WHO African Region. However, many challenges are emerging in the fight against malaria in Africa, such as climate change, poverty, substandard health services and coverage, increased outdoor transmission and the emergence of new vectors, and the growing threat of resistance to antimalarial drugs and insecticides. Joint prevention and treatment, identifying molecular determinants of resistance, new drug development, expanding seasonal malaria chemo-prevention intervention population, and promoting the vaccination of RTS, S/AS01 and R21/Matrix-M may help to solve the dilemma. China's experience in eliminating malaria is conducive to Africa's malaria prevention and control, and China-Africa cooperation needs to be constantly deepened and advanced. Our review aims to help the global public develop a comprehensive understanding of malaria in Africa, thereby contributing to malaria control and elimination., (© 2024. The Author(s).)

Robust diagnostic tools and surveillance are crucial for malaria control and elimination efforts. Malaria caused by neglected Plasmodium parasites is often underestimated due to the lack of rapid diagnostic tools that can accurately detect these species. While nucleic-acid amplification technologies stand out as the most sensitive methods for detecting and confirming Plasmodium species, their implementation in resource-constrained settings poses significant challenges. Here, we present a Pan Plasmodium recombinase polymerase amplification lateral flow (RPA-LF) assay, capable of detecting all six human infecting Plasmodium species in low resource settings. The Pan Plasmodium RPA-LF assay successfully detected low density clinical infections with a preliminary limit of detection between 10-100 fg/µl for P. falciparum. When combined with crude nucleic acid extraction, the assay can serve as a point-of-need tool for molecular xenomonitoring. This utility was demonstrated by screening laboratory-reared Anopheles stephensi mosquitoes fed with Plasmodium-infected blood, as well as field samples of An. funestus s.l. and An. gambiae s.l. collected from central Africa. Overall, our proof-of-concept Pan Plasmodium diagnostic tool has the potential to be applied for clinical and xenomonitoring field surveillance, and after further evaluation, could become an essential tool to assist malaria control and elimination., (© 2024. The Author(s).)

Background: The community involvement and the people's knowledge allow detailed information about the distribution, location, and identification of mosquito breeding-sites. Information which is fundamental for their efficient management and elimination. Since participatory mapping has proven to be an effective tool to identify health determinants, the study aimed to apply the methodology to identify and map potential mosquito breeding-sites in Tambai, Nhamatanda, Mozambique., Methods: A study was conducted using an open-question guide. Discussions were held with 94 participants within ten focus groups, selected in collaboration with local community leaders. A thematic content analysis was performed. Descriptive statistics were used to characterize sociodemographic data. Geographic Positioning System (GPS) was used to compare and map potential breeding-sites. Children under 5 years of age who tested positive for malaria, were georeferenced to the maps., Results: Participants were aware of causes and transmission of malaria, no major differences between groups were observed regarding knowledge and identification of principal potential breeding sites. Gender and age determined specific information, number, and diversity of identified potential breeding sites. A total of 125 potential breeding-sites (36 permanent and 89 temporary) were mapped., Conclusions: Several potential mosquito breeding-sites were identified, located throughout the community, often near house conglomerates and malaria cases. Community participatory mapping could be used to identify potential mosquito breeding-sites by the national malaria control programmes to establish an efficient larval surveillance system, while improving community engagement and control strategies., Trial Registration: ClinicalTrials.gov ID: NCT04419766., (© 2024. The Author(s).)

Background: Malaria is the leading cause of hospitalizations and death in Uganda, particularly in children under the age of five. Studies have shown that adherence to the World Health Organization (WHO) guidelines for the management of severe malaria reduces mortality in hospitalized children. This study aimed to determine the impact of targeted interventions on adherence to the WHO severe malaria treatment guidelines in children at a Ugandan hospital as part of a quality improvement initiative., Methods: Interventions included workflow changes, such as obtaining patient blood samples for diagnostic testing by the admitting healthcare provider as well as utilizing patient caregivers to assist nursing staff in timing medications. An additional intervention was the use of an admission checklist sticker. The post-intervention sample was compared to the baseline assessment. The primary outcome was the proportion of patients receiving care consistent with all aspects of the WHO guidelines. Secondary outcomes included the proportion of patients receiving malaria diagnostic testing, those receiving at least 3 doses of artesunate, the timely administration of artesunate, and adherence to other guideline components. Statistical analyses were conducted using GraphPad PRISM 9.0. Comparisons between groups were analysed using Chi-square or Fisher's exact test for categorical variables and Mann-Whitney test for continuous variables., Results: The post-intervention group included 230 patients with a median age of 5 years [4-8], and 58% of patients were male. Adherence to all aspects of the WHO guidelines was achieved in 10% of patients in the post-intervention group compared to 3% of patients in the baseline (P = 0.007). Appropriate malaria diagnostic testing was performed in 85% of patients post-intervention compared to 66% of patients in the baseline (P < 0.0001). Patients in the post-intervention group were more likely to receive the minimum 3 doses of artesunate (86%) than in the baseline (74%) (P = 0.008). Patients in the post-intervention group were more likely to receive artesunate doses on time than in the baseline (dose 2 P = 0.02, dose 3 P = 0.003)., Conclusions: Targeted, low-cost interventions led to improvement in adherence to severe malaria treatment guidelines. The most notable changes were in malaria diagnostic testing and antimalarial administration., (© 2024. The Author(s).)

Zoonotic malaria, caused by Plasmodium knowlesi, Plasmodium cynomolgi, Plasmodium coatneyi, and Plasmodium inui, is a significant global health concern. The gold standard microscopy, while widely used for malaria diagnosis, faces limitations in differentiating between malaria species. Polymerase chain reaction (PCR), despite its accuracy, is characterized by high costs and time-consuming procedures. This study aims to develop and validate a rapid and accurate diagnostic test for detecting four simian Plasmodium species by using loop-mediated isothermal amplification (LAMP). Loop-mediated isothermal amplification is a cost-effective and faster molecular testing alternative for malaria diagnosis. The project involved designing specific primers, testing sensitivity and specificity against various parasites (including human Plasmodium species, protozoa, and helminths), and evaluating the LAMP assay using 60 macaque samples infected with simian Plasmodium. The LAMP assay exhibited a sensitivity profile enabling the detection of P. knowlesi, P. coatneyi, and P. cynomolgi across a concentration gradient from 5 × 108 down to 5 × 105 parasites/µL. Notably, P. inui was detectable at 5 × 108 parasites/µL. Furthermore, the specificity of the primer tailored for the four simian Plasmodium species was proven, as it produced a positive amplification exclusively for the respective target species and generated negative results for nontarget species. The results indicated that the LAMP assay is capable of detecting simian Plasmodium within a short span of 60 minutes, without any false positives from other samples. This new test has the potential to revolutionize malaria diagnosis, surveillance, and control, thereby mitigating the impact of zoonotic malaria in regions of endemicity.

Introduction: According to the World Health Organization, Microscopy is the gold standard for diagnosing malaria. However, the performance of this examination depends on the experience of the microscopist and the level of parasitemia. Thus, molecular biology detection of malaria could be an alternative technique., Aim: evaluate the contribution of molecular biology in detecting imported malaria., Methods: This was a descriptive, prospective study, including all students, from the Monastir region, and foreigners, from countries endemic to malaria. The study period was from September 2020 to April 2021. Each subject was screened for malaria by three methods: direct microscopic detection of Plasmodium, detection of plasmodial antigens, and detection of plasmodial DNA by nested PCR., Results: Among the 127 subjects screened, only one had a positive microscopic examination for Plasmodium falciparum. Among the 126 subjects with a negative microscopic examination, twelve students had a positive nested PCR result, i.e. 9.5%. Molecular sequencing allowed the identification of ten isolates of Plasmodium falciparum, one Plasmodium malariae and one Plasmodium ovale. Our study showed that the results of nested PCR agreed with those of microscopy in 90.6% of cases., Conclusion: Nested PCR seems more sensitive for the detection of low parasitemias. Hence the importance of including molecular biology as a malaria screening tool to ensure better detection of imported cases.

Malaria is a significant health concern worldwide, particularly in Africa where its prevalence is still alarmingly high. Using artificial intelligence algorithms to diagnose cells with malaria provides great convenience for clinicians. In this paper, a densely connected convolutional dynamic learning network (DCDLN) is proposed for the diagnosis of malaria disease. Specifically, after data processing and partitioning of the dataset, the densely connected block is trained as a feature extractor. To classify the features extracted by the feature extractor, a classifier based on a dynamic learning network is proposed in this paper. Based on experimental results, the proposed DCDLN method demonstrates a diagnostic accuracy rate of 97.23%, surpassing the diagnostic performance than existing advanced methods on an open malaria cell dataset. This accurate diagnostic effect provides convincing evidence for clinicians to make a correct diagnosis. In addition, to validate the superiority and generalization capability of the DCDLN algorithm, we also applied the algorithm to the skin cancer and garbage classification datasets. DCDLN achieved good results on these datasets as well, demonstrating that the DCDLN algorithm possesses superiority and strong generalization performance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Malaria remains a significant global public health concern, with a recent increase in the number of zoonotic malaria cases in Southeast Asian countries. However, limited reports on the vector for zoonotic malaria exist owing to difficulties in detecting parasite DNA in Anopheles mosquito vectors. Herein, we demonstrate for the first time that several Anopheles mosquitoes contain simian malaria parasite DNA using droplet digital PCR (ddPCR), a highly sensitive PCR method. An entomological survey was conducted to identify simian malaria vector species at Phra Phothisat Temple (PPT), central Thailand, recognized for a high prevalence of simian malaria in wild cynomolgus macaques. A total of 152 mosquitoes from six anopheline species were collected and first analyzed by a standard 18S rRNA nested-PCR analysis for malaria parasite which yielded negative results in all collected mosquitoes. Later, ddPCR was used and could detect simian malaria parasite DNA, i.e. Plasmodium cynomolgi, in 25 collected mosquitoes. And this is the first report of simian malaria parasite DNA detection in Anopheles sawadwongporni. This finding proves that ddPCR is a powerful tool for detecting simian malarial parasite DNA in Anopheles mosquitoes and can expand our understanding of the zoonotic potential of malaria transmission between monkeys and humans., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Need for affordable, rapid and user-friendly point of care (POC) devices are increasing exponentially for strengthening the health care system in primary care as well as for self- monitoring in routine analysis. In addition to routine analysis of glucose, Covid-19 type fast spreading, infectious diseases have created further push for exploring rapid, cost-effective and self-monitoring diagnostic devices. Successful implementation of self-monitoring devices for Covid -19 has been realized. However, not much success has been realized for malaria and dengue which are two fatal diseases that affect the population in underdeveloped and developing countries. To monitor the presence of parasites for these diseases, rapid, onsite monitoring devices are still being explored. In this review, we present a review of the research carried out on electrochemical POC devices for monitoring infectious diseases such as Covid-19, malaria and dengue., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

For over a century, the need to identify malaria in the peripheral blood has been the driving force behind the development of fundamental clinical microscopy techniques. In the study by Moysis et al., artificial intelligence-based model was utilized to identify and provide quantitative morphological characteristics of red blood cells typical to severe malaria anaemia, irrespective to the actual presence of visible parasites. Commentary on: Moysis et al. Leveraging deep learning for detecting red blood cell morphological changes in blood films from children with severe malaria anaemia. Br J Haematol 2024;205:699-710., (© 2024 The Author(s). British Journal of Haematology published by British Society for Haematology and John Wiley & Sons Ltd.)

Private medicine retailers (PMRs) such as pharmacies and drug stores account for a substantial share of treatment-seeking for fever and malaria, but there are widespread concerns about quality of care, including inadequate access to malaria rapid diagnostic tests (RDTs) and artemisinin-based combination therapies (ACTs). This review synthesizes evidence on the effectiveness of interventions to improve malaria case management in PMRs in sub-Saharan Africa (PROSPERO #2021:CRD42021253564). We included quantitative studies evaluating interventions supporting RDT and/or ACT sales by PMR staff, with a historical or contemporaneous control group, and outcomes related to care received. We searched Medline Ovid, Embase Ovid, Global Health Ovid, Econlit Ovid and the Cochrane Library; unpublished studies were identified by contacting key informants. We conducted a narrative synthesis by intervention category. We included 41 papers, relating to 34 studies. There was strong evidence that small and large-scale ACT subsidy programmes (without RDTs) increased the market share of quality-assured ACT in PMRs, including among rural and poorer groups, with increases of over 30 percentage points in most settings. Interventions to introduce or enhance RDT use in PMRs led to RDT uptake among febrile clients of over two-thirds and dispensing according to RDT result of over three quarters, though some studies had much poorer results. Introducing Integrated Community Case Management (iCCM) was also effective in improving malaria case management. However, there were no eligible studies on RDT or iCCM implementation at large scale. There was limited evidence that PMR accreditation (without RDTs) increased ACT uptake. Key evidence gaps include evaluations of RDTs and iCCM at large scale, evaluations of interventions including use of digital technologies, and robust studies of accreditation and other broader PMR interventions., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Goodman et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: In countries where malaria is endemic, the use of rapid diagnostic tests(RDTs) has become routine, especially in rural settings. Such regions are characterised by often having other co-endemic infectious diseases, at high levels of prevalence., Aim: To illustrate the potential added-value of "sentinel" screening for patients presenting for a routine diagnostic test for malaria, at healthcare facilities in Uganda., Methods: We developed an economic model by combining two decision trees, one for malaria and a second for the co-endemic disease schistosomiasis. The integrated model was designed to inform policy strategies for the co-endemic disease in addition to malaria (i.e., whether to test opportunistically for schistosomiasis or use mass drug administration(MDA) as per usual practice).We performed the analysis on three comparators varying testing accuracy and costs., Results: Sentinel screening can provide added value to the testing of patients compared with the status quo: when schistosomiasis prevalence is high then MDA is preferential; if low prevalence, treating no one is preferred. If the disease has average levels of prevalence, then a strategy involving testing is preferred. Prevalence thresholds driving the dominant strategy are dependent upon the model parameters, which are highly context specific. At average levels of prevalence for schistosomiasis and malaria for Uganda, adding a sentinel screening was cost-effective when the accuracy of test was higher than current diagnostics and when economies of scope were generated(Expected value clinical Information = 0.65$ per DALY averted, 137.91$ per correct diagnoses).Protocols using diagnostics with current accuracy levels were preferred only for levels of MDA coverage below 75%., Conclusion: The importance of the epidemiological setting is crucial in determining the best cost-effective strategy for detecting endemic disease. Economies of scope can make sentinel screenings cost-effective strategies in specific contexts. Blanket thresholds recommended for MDA may not always be the preferred option for endemic diseases., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Manca et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

To evaluate the clinical significance of PLT, MPV, and PDW in monitoring malaria treatment efficacy and predicting disease progression. A total of 31 patients with imported malaria were selected as the observation group, while 31 non-malaria patients with fever were selected as controls. The observation group was subdivided into a complication group and a non-complication group according to the occurrence of complications during treatment. Additionally, on the 1st day (within 24 h), the 3rd day, and the 5th day following admission, a comprehensive blood routine examination, Plasmodium microscopic examination, and colloidal gold assay were conducted. The blood routine examination results were compared before and after treatment among patients in the observation group and the control group. Moreover, the study involved dynamic monitoring and analysis of the levels and variations in PLT, MPV, and PDW within both the complication group and the non-complication group. The Plasmodium density was negatively correlated with PLT before treatment. There were significant differences were observed in PLT, MPV, and PDW (P < 0.05) within the observation group before and after treatment. Notably, there were no significant alterations in red blood cell (RBC), hemoglobin (Hb), and white blood cell (WBC) counts (P > 0.05) within the observation group before and after treatment. The PLT, MPV, and PDW levels in the complication group and the non-complication group exhibited an upward trend after treatment. Further, the PLT of patients in the complication group was significantly lower than that in the non-complication group. Additionally, the PLT, MPV, and PDW levels in the complication group and the non-complication group increased gradually from the time of admission to the 3rd and 5th day of treatment. Notably, the PLT in the complication group was consistently lower than that in the non-complication group. The continuous monitoring of PLT, MPV, and PDW changes plays a crucial role in assessing malaria treatment efficacy and prognosis in these individuals., (© 2024. The Author(s).)

Malaria continues to be a major source of morbidity and mortality in sub-Saharan Africa. Timely, accurate, and effective case management is critical to malaria control. Proactive community case management (ProCCM) is a new strategy in which a community health worker "sweeps" a village, visiting households at defined intervals to proactively provide diagnostic testing and treatment if indicated. Pilot experiments have shown the potential of ProCCM for controlling malaria transmission; identifying the best strategy for administering ProCCM in terms of interval timings and number of sweeps could lead to further reductions in malaria infections. We developed an agent-based simulation to model malaria transmission and the impact of various ProCCM strategies. The model was validated using symptomatic prevalence data from a ProCCM pilot study in Senegal. Various ProCCM strategies were tested to evaluate the potential for reducing parasitologically confirmed symptomatic malaria cases in the Senegal setting. We found that weekly ProCCM sweeps during a 21-week transmission season could reduce cases by 36.3% per year compared with no sweeps. Alternatively, two initial fortnightly sweeps, seven weekly sweeps, and finally four fortnightly sweeps (13 sweeps total) could reduce confirmed malaria cases by 30.5% per year while reducing the number of diagnostic tests and corresponding costs by about 33%. Under a highly seasonal transmission setting, starting the sweeps early with longer duration and higher frequency would increase the impact of ProCCM, though with diminishing returns. The model is flexible and allows decision-makers to evaluate implementation strategies incorporating sweep frequency, time of year, and available budget.

Background: Togo's National Malaria Control Programme has initiated an active home-based malaria management model for all age groups in rural areas of Bassar Health District. This report describes the model, reports its main results, and determines the factors associated with positive rapid diagnostic test results., Methods: From 2014 to 2017, in three peripheral care units of Bassar Health District (Binaparba, Nangbani, and Baghan), community health workers visited residents' homes weekly to identify patients with malaria symptoms, perform rapid diagnostic tests in symptomatic patients, and give medication to positive cases. Univariate and multivariate logistic regression models were used to determine the factors associated with positive tests., Results: The study covered 11,337 people (817 in 2014, 1804 in 2015, 2638 in 2016, and 6078 in 2017). The overall mean age was 18 years (95% CI 5-29; min-max: 0-112 years). The median age was 10 years (SD: 16.9). The proportions of people tested positive were 75.3% in Binaparba, 77.4% in Nangbani, and 56.6% in Baghan. The 5-10 age group was the most affected category (24.2% positive tests). Positive tests were more frequent during the rainy than during the dry season (62 vs. 38%) and the probability of positive test was 1.76 times higher during the rainy than during the dry season (adjusted OR = 1.74; 95% CI 1.60-1.90). A fever (37.5 °C or higher) increased significantly the probability of positive test (adjusted OR = 2.19; 95% CI 1.89-2.54). The risk of positive test was 1.89 times higher in passive than in active malaria detection (adjusted OR = 1.89; 95% CI 1.73-2.0)., Conclusions: This novel experimental community and home-based malaria management in Togo suggested that active detection of malaria cases is feasible within 24 h, which allows rapid treatments before progression to often-fatal complications. This PECADOM + program will help Togo's National Malaria Control Programme reduce malaria morbidity and mortality in remote and hard-to-reach communities., (© 2024. The Author(s).)