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Purpose: The current diagnostic methods for leptospirosis diagnosis are technically complex and expensive, with limited applicability to specialized laboratories. Furthermore, they lack diagnostic accuracy in the acute stage of the disease, which coincides with a period when antibiotics are highly effective. New simple and accurate tests are mandatory to decentralize and improve diagnosis. Here, we introduced a new lateral flow immunoassay (Lepto-LF) for human leptospirosis. Methods: We conducted a double-blinded assay using 104 serum samples from patients with confirmed or discarded diagnosis for leptospirosis. The diagnostic performance of Lepto-LF was estimated across different ranges of days from onset of symptoms (dpo), considering the diagnostic algorithm as reference standard. Additionally, it was compared with the screening methods enzyme-linked immunosorbent assay (IgM-ELISA) and the slide agglutination test using temperature-resistant antigen (SATR). Results: Lepto-LF exhibited perfect diagnostic performance with a Youden´s index J = 1 from 6 dpo in the acute phase. IgM-ELISA gave slightly lower accuracy with J = 0.91 and 95.5% of both sensitivity and specificity; while SATR showed a markedly inferior yield (J = 0.41, sensitivity = 95.5%, specificity = 45.5%). The performances remained consistent in the convalescence phase of the disease (> 10 dpo). Conclusion: Lepto-LF was found to be a reliable test for simple, rapid and early diagnosis of leptospirosis, resulting a promising tool for decentralizing leptospirosis diagnosis and enabling timely treatment of patients. In addition, Lepto-LF may be employed as confirmatory test, especially in remote areas and vulnerable contexts where the standard MAT is not available. [ABSTRACT FROM AUTHOR]

Objectives: To develop a system for the rapid detection of Acinetobacter baumannii 173-p1 antibiotic resistance (to ensure reliable fixation of bacteria on a cantilever without losing their nanomotion, to show that nanomotion is due to bacterial metabolism, to compare the nanomotion of bacteria in suspension form and inside of the biofilms), to study the sensitivity/resistance of A. baumannii 173-p1 to antibiotics (lincomycin, ceftriaxone and doxycycline) using the oscillation method of atomic force microscopy and to evaluate the sensitivity and speed of the method in comparison with the classical disk diffusion method. Methods: The oscillation mode of atomic force microscopy, scanning electron microscopy and the classical disk diffusion method were used for a complex parallel study of A. baumannii 173-p1 antibiotic resistance, which included testing of fixing agents (poly-L-lysine, rosin and fibronectin), comparison of bacterial metabolism in a set of media (normal saline solution, meat-peptone broth and lysogeny broth) and assessment of antibiotic sensitivity/resistance per se. Results: A method for express testing of Acinetobacter baumannii antibiotic resistance using AFM was developed; it is shown that bacterial nanomotion directly correlates with bacteria metabolic activity and that bacterial nanomotion is more easily detected in suspension form, rather than in biofilms. Conclusion: The express testing method gave results that are completely comparable with the classical disk diffusion test and with the results of morphology studies by the SEM method, but it significantly exceeded them in speed, allowing a conclusion to be made on the sensitivity/resistance of bacteria less than an hour after the start of the diagnostics. [ABSTRACT FROM AUTHOR]

Introduction: In the fight against virus-caused pandemics like COVID-19, diagnostic tests based on RT-qPCR are essential, but they are sometimes limited by their dependence on expensive, specialized equipment and skilled personnel. Consequently, an alternative nucleic acid detection technique that overcomes these restrictions, called loop-mediated isothermal amplification following reverse transcription (RTLAMP), has been broadly investigated. Nevertheless, the developed RT-LAMP assays for SARS-CoV-2 detection still require laboratory devices and electrical power, limiting their widespread use as rapid home tests. This work developed a flexible RT-LAMP assay that gets beyond the drawbacks of the available isothermal LAMP-based SARS-CoV-2 detection, establishing a simple and effective at-home diagnostic tool for COVID-19. Methodology: A multiplex direct RT-LAMP assay, modified from the previously developed test was applied to simultaneously identify the two genes of SARS-CoV-2. We used a colorimetric readout, lyophilized reagents, and benchmarked an electro-free and micropipette-free method that enables sensitive and specific detection of SARS-CoV-2 in home settings. Results: Forty-one nasopharyngeal swab samples were tested using the developed home-testing RT-LAMP (HT-LAMP) assay, showing 100% agreement with the RT-qPCR results. Conclusions: This is the first electrically independent RT-LAMP assay successfully developed for SARS-CoV-2 detection in a home setting. Our HT-LAMP assay is thus an important development for diagnosing COVID-19 or any other infectious pandemic on a population scale. [ABSTRACT FROM AUTHOR]

Background: Respiratory infections pose a major public health threat. The predominant viruses causing viral respiratory infections are influenza A and B (Flu-A, Flu-B), coronaviruses, respiratory syncytial virus (RSV), and adenovirus. This study aims to investigate the proportion of these cases via rapid antigen tests and assess seasonal patterns. Methods: Clinical samples were collected from symptomatic adults presenting to the Emergency and Respiratory Medicine Departments of the University Hospital of Larissa (UHL), Greece from 16 October 2023 to 31 March 2024. Nasal specimens were antigen-tested for Flu-A/B, SARS-CoV-2, RSV, and adenovirus. Results: The total sample of specimens collected was 1434, of which 739 (51.5%) were female and 695 were male (48.5%). The mean age of participants was 57 ± 5.5 years. Among the positive results, we recorded a proportion of 40.18% and 11.40% for influenza A and B, respectively, followed by 35.79% for SARS-CoV-2, 10.70% for RSV, and 1.93% for adenovirus. Conclusions: In Greece, surveillance systems in infection control are underutilized. Rapid tests via multiple antigens can quickly identify viral infections, making them a valuable tool with financial benefits for health systems. Early detection of respiratory infections helps allocate resources efficiently, ensures adequate staff and facilities are available, and improves patient care through refined clinical management. [ABSTRACT FROM AUTHOR]

We meta-analyzed the diagnostic accuracy of rapid diagnostic tests (dipsticks) and loop-mediated isothermal amplification (LAMP) method to detect Shigella species.We searched MEDLINE, Embase, Web of Science and Google Scholar from inception to 2023 for studies reporting on the performance of Shigella dipstick and LAMP tests compared with culture or polymerase chain reaction (PCR).Our search identified 2618 studies, of which fourteen met the inclusion criteria for the systematic review. Ten studies covering 4056 tests (from twelve countries) were included in the meta-analysis. The overall pooled sensitivity and specificity were 98% (95% CI: 94–100) and 97% (95% CI: 92–99), respectively. Pooled sensitivity and specificity of dipsticks were 95% and 98%, respectively. In contrast, LAMP showed higher pooled sensitivity (100%) and diagnostic odds ratio (431752), but similar specificity (97%).LAMP and dipstick tests exhibited promising performance, suggesting that they could be useful for assisting in the diagnosis of shigellosis.

Feline parvovirus (FPV) is one of the most common causes of acute hemorrhagic enteritis, with high mortalities in kittens. As clinical diagnosis is often indecisive, the aim of our study was to evaluate the diagnostic accuracy of in-clinic rapid tests for the detection of FPV infection in cats. To this end, we contrasted the most reliable diagnostic technique--PCR--with the rapid diagnostic kit for FPV antigen. A total of 100 stool samples were collected from cats suspected of being infected with FPV. A vaccination history was reported for all sampled cats. Every sample underwent both PCR and the rapid diagnostic test, with the results being compared. Anorexia, bloody diarrhea, severe dehydration, hypothermia, and vomiting were the most common clinical findings significantly associated with parvovirus- infected cats. 35 out of 100 clinically ill cats were FPV positive (35%) using the rapid screening test, while 43 (43%) of the tested samples were PCR positive. Overall, the two tests found FPV infection in 32 cases and ruled it out in 54. The PCR technique confirmed the infection in 11 cases that their rapid testing were negative. Rapid antigen-based screening assays demonstrated sensitivity and specificity of 74.42% and 94.74%, respectively. In conclusion, a positive result certainly indicates FPV infection, while a negative result does not rule out parvoenteritis from the differential diagnosis, especially in cats exhibiting clinical symptoms. It is possible to anticipate increased sensitivity if the test is run right following sample collection. [ABSTRACT FROM AUTHOR]

Rapid tests allow outpatient, low cost, reliable, screening for chronic HIV infection. However, data regarding their sensitivity on primary infection remain scarce. The objective of this study was to assess sensitivity of nine HIV rapid tests for primary HIV-1 infection screening. Seventy-five serum samples from patients during HIV-1 primary infection were included. Primary infection was diagnosed by a positive 4th generation ELISA and HIV-1 RNA positivity confirmed by Western blot patterns associated with HIV-1 primary infection. Early seroconversion was defined as the absence of antibodies on HIV-1 Western blot associated with HIV-1 RNA and p24-antigen positivity. An identical sensitivity (95% CI) of 76.7% (65.2–84.2%) was observed for HIV 1/2 STAT-PAK® Assay (STAT-PAK), INSTI™ HIV-1/HIV-2 antibody Test (INSTI), SURE CHECK® HIV 1/2 (SURE CHECK) and MULTISURE HIV rapid test (MULTISURE) with visual reading. Sensitivity was 74.7% (63.8–83.1%) for MULTISURE (automatic reading), 77.0% (66.3–85.1%) for FIRST RESPONSE® Test VIH 1–2.O CARTE (FIRST RESPONSE), 83.8% (73.8–90.5%) for VIKIA HIV1/2® (VIKIA), 88.0% (78.7–93.6%) for Genie™ Fast HIV 1/2 (Genie Fast), 88.6% (79.0-94.1%) for Hexagon HIV (Hexagon), and 92.8% (83.6–96.3%) for Exacto® TEST HIV Pro (Exacto). However, rapid tests performed poorly for the early seroconversion subgroup (n = 14), with sensitivities ranging from 7% (1.3–31.5%) for STAT-PAK, INSTI, SURE CHECK, MULTISURE (automatic reading), to 29% (12–55%) for FIRST RESPONSE, 31% (13–58%) for VIKIA, 43% (21–67%) for Hexagon and 57.1% (32.6–78.6%) for Exacto and Genie Fast. Overall, despite significant discrepancies in sensitivity, HIV rapid tests should be used with caution in the context of a suspected primary infection. [ABSTRACT FROM AUTHOR]

We meta-analyzed the diagnostic accuracy of rapid diagnostic tests (dipsticks) and loop-mediated isothermal amplification (LAMP) method to detect Shigella species. We searched MEDLINE, Embase, Web of Science and Google Scholar from inception to 2023 for studies reporting on the performance of Shigella dipstick and LAMP tests compared with culture or polymerase chain reaction (PCR). Our search identified 2618 studies, of which fourteen met the inclusion criteria for the systematic review. Ten studies covering 4056 tests (from twelve countries) were included in the meta-analysis. The overall pooled sensitivity and specificity were 98 % (95 % CI: 94–100) and 97 % (95 % CI: 92–99), respectively. Pooled sensitivity and specificity of dipsticks were 95 % and 98 %, respectively. In contrast, LAMP showed higher pooled sensitivity (100 %) and diagnostic odds ratio (431752), but similar specificity (97 %). LAMP and dipstick tests exhibited promising performance, suggesting that they could be useful for assisting in the diagnosis of shigellosis. • Shigella infection can result in growth deficits among children. • We investigated the diagnostic accuracy of Shigella dipsticks tests and LAMP. • Ten studies were included in the meta-analysis. • Dipsticks pooled sensitivity and specificity: 95 % and 98 %, respectively. • LAMP pooled sensitivity and specificity: 100 % and 97 %, respectively. [ABSTRACT FROM AUTHOR]

The identification of the type of body fluid in crime scene evidence may be crucial, so that the efforts are high to reduce the complexity of these analyses and to minimize time and costs. Reliable immunochromatographic rapid tests for specific and sensitive identification of blood, saliva, urine and sperm secretions are already routinely used in forensic genetics. The recently introduced Seratec® PMB test is said to detect not only hemoglobin, but also differentiate menstrual blood from other secretions containing blood (cells) by detecting D-dimers. In our experimental set-up, menstrual blood could be reliably detected in mock forensic samples. Here, the result was independent of sample age and extraction buffer volume. It was also successfully demonstrated that all secretions without blood cells were negative for both, hemoglobin (P) and D-dimer (M). However, several blood cell–containing secretions/tissues comprising blood (injury), nasal blood, postmortem blood and wound crust also demonstrated positive results for D-dimer (M) and were therefore false positives. For blood (injury) and nasal blood, this result was reproduced for different extraction buffer volumes. The results of this study clearly demonstrate that the Seratec® PMB test is neither useful nor suitable for use in forensic genetics because of the great risk of false positive results which can lead to false conclusions, especially in sexual offense or violent acts. [ABSTRACT FROM AUTHOR]

Abstract Background While Plasmodium falciparum and Plasmodium vivax cause the majority of malaria cases and deaths, infection by Plasmodium malariae and other Plasmodium species also causes morbidity and mortality. Current understanding of these infections is limited in part by existing point-of-care diagnostics that fail to differentiate them and have poor sensitivity for low-density infections. Accurate diagnosis currently requires molecular assays performed in well-resourced laboratories. This report describes the development of a P. malariae diagnostic assay that uses rapid, isothermal recombinase polymerase amplification (RPA) and lateral-flow-strip detection. Methods Multiple combinations of custom RPA primers and probes were designed using publicly available P. malariae genomic sequences, and by modifying published primer sets. Based on manufacturer RPA reaction conditions (TwistDx nfo kit), an isothermal assay was optimized targeting the multicopy P. malariae 18S rRNA gene with 39 °C incubation and 30-min run time. RPA product was visualized using lateral strips (FAM-labeled, biotinylated amplicon detected by a sandwich immunoassay, visualized using gold nanoparticles). Analytical sensitivity was evaluated using 18S rRNA plasmid DNA, and clinical sensitivity determined using qPCR-confirmed samples collected from Tanzania, Ethiopia, and the Democratic Republic of the Congo. Results Using 18S rRNA plasmid DNA, the assay demonstrates a detection limit of 10 copies/µL (~ 1.7 genome equivalents) and 100% analytical specificity. Testing in field samples showed 95% clinical sensitivity and 88% specificity compared to qPCR. Total assay time was less than 40 min. Conclusion Combined with simplified DNA extraction methods, the assay has potential for future field-deployable, point-of-care use to detect P. malariae infection, which remains largely undiagnosed but a neglected cause of chronic malaria. The assay provides a rapid, simple readout on a lateral flow strip without the need for expensive laboratory equipment.

Abstract Compared to other infectious diseases, for which LFT development can take years, SARS-CoV-2 antigen LFTS were developed and deployed within months. LFTS for antigen detection were adopted on an unprecedented scale during the COVID-19 pandemic, but many of them lack the sensitivity especially for samples with low viral load. In our previous work, we developed an enhanced signal strip for detection of SARS CoV-2 SI antigens in saliva. Here we introduce some modification to improve the sensitivity, and specificity, and to lower the cost of the strip, by using biotin streptavidin (BS) system. In the modified BS strip, gold-streptavidin and biotinylated Nanobodies (Nbs) against S1 antigen were externally mixed with the tested samples (saliva or nasopharyngeal swab) before their application on the sample pad of the test strip containing angiotensin converting enzyme (ACE-2), as the capturing probe. The study included 320 individuals, with 180 being positively confirmed by RT-PCR and 140 confirmed negative, as well as, 45 health care workers, who were responsible for screening and handling of surgical cases in General Surgery Department and COVID clinic of TBRI. Our results proved that modified BS strip improved the overall sensitivity and specificity of S1antigen detection in saliva samples (95.21% and 99.29% respectively) compared to our previously developed enhanced LFTS (91.66% and 98.57% respectively). Also, the sensitivity of cases with Ct ≤ 30, Ct $$\le$$ ≤ 35, and Ct $$\le$$ ≤ 40 using the modified BS strip showed higher values (98.54%, 95.38%, and 88.89% respectively), compared to the corresponding results of our previously developed enhanced LFTS (95.86%, 92.31%, and 82.22% respectively). There were no cross-reactions with either Middle East respiratory syndrome corona virus MERS-CoV or SARS-CoV antigens. Furthermore, we found that the lower viral detection limit (LVD) of BS strip was obviously lower than our previous LVD limit of the enhanced LFTS (0.2 × 104 copies/ml vs. 0.4 × 104 copies/ml, respectively). Our developed BS strip showed that saliva samples gave better results than nasopharyngeal swabs of the same patients. The fact of using smaller amounts of Nbs, and ACE2, as well as the dispensing off of conjugate pad when applying BS strip modifications, justified the expected reduction in the costs of the strip. The implementation of BS strips on saliva samples of 45 health co-workers, who were tested 4 and 6 days after exposure to infection, showed an increase in the sensitivity, starting from the 4th day and reaching its highest level on the 6th day in both high risk and paramedic groups (90.9%, and 80.0%, respectively). This study provides evidence that employment of the modified BS system could increase the sensitivity of the strips, lower their cost, and render them an effective screening tool for early detection of the virus in saliva of suspected Covid-19 patients.

Background: While Plasmodium falciparum and Plasmodium vivax cause the majority of malaria cases and deaths, infection by Plasmodium malariae and other Plasmodium species also causes morbidity and mortality. Current understanding of these infections is limited in part by existing point-of-care diagnostics that fail to differentiate them and have poor sensitivity for low-density infections. Accurate diagnosis currently requires molecular assays performed in well-resourced laboratories. This report describes the development of a P. malariae diagnostic assay that uses rapid, isothermal recombinase polymerase amplification (RPA) and lateral-flow-strip detection. Methods: Multiple combinations of custom RPA primers and probes were designed using publicly available P. malariae genomic sequences, and by modifying published primer sets. Based on manufacturer RPA reaction conditions (TwistDx nfo kit), an isothermal assay was optimized targeting the multicopy P. malariae 18S rRNA gene with 39 °C incubation and 30-min run time. RPA product was visualized using lateral strips (FAM-labeled, biotinylated amplicon detected by a sandwich immunoassay, visualized using gold nanoparticles). Analytical sensitivity was evaluated using 18S rRNA plasmid DNA, and clinical sensitivity determined using qPCR-confirmed samples collected from Tanzania, Ethiopia, and the Democratic Republic of the Congo. Results: Using 18S rRNA plasmid DNA, the assay demonstrates a detection limit of 10 copies/µL (~ 1.7 genome equivalents) and 100% analytical specificity. Testing in field samples showed 95% clinical sensitivity and 88% specificity compared to qPCR. Total assay time was less than 40 min. Conclusion: Combined with simplified DNA extraction methods, the assay has potential for future field-deployable, point-of-care use to detect P. malariae infection, which remains largely undiagnosed but a neglected cause of chronic malaria. The assay provides a rapid, simple readout on a lateral flow strip without the need for expensive laboratory equipment. [ABSTRACT FROM AUTHOR]

Compared to other infectious diseases, for which LFT development can take years, SARS-CoV-2 antigen LFTS were developed and deployed within months. LFTS for antigen detection were adopted on an unprecedented scale during the COVID-19 pandemic, but many of them lack the sensitivity especially for samples with low viral load. In our previous work, we developed an enhanced signal strip for detection of SARS CoV-2 SI antigens in saliva. Here we introduce some modification to improve the sensitivity, and specificity, and to lower the cost of the strip, by using biotin streptavidin (BS) system. In the modified BS strip, gold-streptavidin and biotinylated Nanobodies (Nbs) against S1 antigen were externally mixed with the tested samples (saliva or nasopharyngeal swab) before their application on the sample pad of the test strip containing angiotensin converting enzyme (ACE-2), as the capturing probe. The study included 320 individuals, with 180 being positively confirmed by RT-PCR and 140 confirmed negative, as well as, 45 health care workers, who were responsible for screening and handling of surgical cases in General Surgery Department and COVID clinic of TBRI. Our results proved that modified BS strip improved the overall sensitivity and specificity of S1antigen detection in saliva samples (95.21% and 99.29% respectively) compared to our previously developed enhanced LFTS (91.66% and 98.57% respectively). Also, the sensitivity of cases with Ct ≤ 30, Ct ≤ 35, and Ct ≤ 40 using the modified BS strip showed higher values (98.54%, 95.38%, and 88.89% respectively), compared to the corresponding results of our previously developed enhanced LFTS (95.86%, 92.31%, and 82.22% respectively). There were no cross-reactions with either Middle East respiratory syndrome corona virus MERS-CoV or SARS-CoV antigens. Furthermore, we found that the lower viral detection limit (LVD) of BS strip was obviously lower than our previous LVD limit of the enhanced LFTS (0.2 × 104 copies/ml vs. 0.4 × 104 copies/ml, respectively). Our developed BS strip showed that saliva samples gave better results than nasopharyngeal swabs of the same patients. The fact of using smaller amounts of Nbs, and ACE2, as well as the dispensing off of conjugate pad when applying BS strip modifications, justified the expected reduction in the costs of the strip. The implementation of BS strips on saliva samples of 45 health co-workers, who were tested 4 and 6 days after exposure to infection, showed an increase in the sensitivity, starting from the 4th day and reaching its highest level on the 6th day in both high risk and paramedic groups (90.9%, and 80.0%, respectively). This study provides evidence that employment of the modified BS system could increase the sensitivity of the strips, lower their cost, and render them an effective screening tool for early detection of the virus in saliva of suspected Covid-19 patients. [ABSTRACT FROM AUTHOR]

African swine fever (ASF) is a contagious disease of wild boar and domestic pigs notifiable to the World Organisation for Animal Health due to its high socio-economic impact. ASF is caused by the complex ASF virus (ASFV), and it can present different clinical manifestations that can be confused with other diseases; for this reason, laboratory testing is necessary for the proper diagnosis of clinically suspected animals. Despite the efforts put into it over decades, no treatment or safe vaccine is globally available, and disease control is based on early diagnosis and the implementation of strict biosecurity measures. In this context, rapid tests have the potential to accelerate and facilitate the identification of infected animals by giving fast on-site results. In this work, we improved the available point-of-care assays for the diagnosis of the disease by the development of a more specific antigen test and a more sensitive antibody test. This antibody detection test allowed for the earlier detection of infected animals than two commercial indirect ELISAs (statistically significant). Moreover, we developed a combined dual rapid test, unifying, in the same cassette, an antigen detection strip and an antibody detection strip. In this study, we confirmed that this combo approach is a useful tool for implementing rapid tests in the field since it increases the percentage of positive samples detected, even when PCR turns negative, while maintaining a good specificity. [ABSTRACT FROM AUTHOR]

Food and fish adulteration is a major public concern worldwide. Apart from economic fraud, health issues are in the forefront mainly due to severe allergies. Sardines are one of the most vulnerable-to-adulteration fish species due to their high nutritional value. Adulteration comprises the substitution of one fish species with similar species of lower nutritional value and lower cost. The detection of adulteration, especially in processed fish products, is very challenging because the morphological characteristics of the tissues change, making identification by the naked eye very difficult. Therefore, new analytical methods and (bio)sensors that provide fast analysis with high specificity, especially between closely related fish species, are in high demand. DNA-based methods are considered as important analytical tools for food adulteration detection. In this context, we report the first DNA sensors for sardine species identification. The sensing principle involves species recognition, via short hybridization of PCR-amplified sequences with specific probes, capture in the test zone of the sensor, and detection by the naked eye using gold nanoparticles as reporters; thus, avoiding the need for expensive instruments. As low as 5% adulteration of Sardina pilchardus with Sardinella aurita was detected with high reproducibility in the processed mixtures simulating canned fish products. [ABSTRACT FROM AUTHOR]

Background Human immunodeficiency virus (HIV) rapid diagnostic tests (RDTs) are widely used. However, buffer stockouts commonly lead to utilising non-approved liquids, resulting in errors. Our aim was to evaluate the diagnostic accuracy of an alternative buffer. Methods Paired Determine HIV-1/2 rapid tests with commercial buffer and locally produced 0.01M phosphate-buffered saline (PBS) were performed on consecutive consenting individuals requiring HIV testing. Serum samples were sent for confirmation through the local gold-standard algorithm (Murex HIV Ag/Ab, Hexagon HIV with/without Geenius HIV 1/2). Test accuracy, κ and exact McNemar's test were also carried out. Results Of 167 participants, 137 had confirmatory testing. The sensitivity of the Determine HIV-1/2 test using PBS compared with the gold standard was 100% (95% confidence interval [CI] 90.5 to 100) with a specificity of 98% (95% CI 92.9 to 99.8). The κ value was 0.94 compared with the gold standard and 0.92 compared with the Determine HIV-1/2 test using the commercial buffer. McNemar's test showed no evidence of differing sensitivities. Due to operational constraints, the study included 37 of the 49 positive cases as determined by the sample size calculation, resulting in an attained power of 80% instead of the intended 90%. Conclusions These results suggest that 0.01M PBS is an alternative solution for Determine HIV-1/2 when buffer stockouts occur. [ABSTRACT FROM AUTHOR]

Helicobacter pylori (HPY) is a type of spiral bacterium. The International Agency for Research on Cancer (IARC) has classified HPY as a Group 1 carcinogen, indicating that it is the leading cause of stomach cancer. The traditional testing method for HPY is cumbersome and requires professional personnel to operate. Owing to advances in biochemical technology, rapid tests for HPY have been developed. The development of the rapid test is based on colloidal gold. The rapid test displays two lines when the result is positive. The color of the test result can vary due to changes in the size or concentration of colloidal particles. Users may misinterpret the results owing to the effect of ambient lighting or when the color is too close to the background color. In this study, we successfully developed a method for using a smartphone to evaluate the results and sample concentration in rapid tests for detecting HPY. The HPY rapid test is placed on a colorimetric board developed for this study, and an image is captured using a smartphone and evaluated using the image processing program designed in this research. The ArUco marker is used for positioning and color correction to ensure that it is not affected by the brand of the camera of the mobile phone and nonuniform ambient light sources. To confirm whether the proposed method correctly determines the result and sample concentration, experiments were conducted using multiple different mobile phones. In the experiment, the illumination and shooting height were fixed and several different sample concentrations were used. Printed simulated and real rapid test results were used in the experiment. The experimental results show that both positive and negative can be correctly determined, and there is good linearity when determining each concentration of samples from the image. For example, when using simulated rapid tests, the R² value is at least 0.9915, and the detection concentration threshold of the real HPY rapid tests is 5000 CFU/mL. Even with the concentration at 1250 CFU/mL, the R² value is still 0.9473. [ABSTRACT FROM AUTHOR]

In this study, an impedance electrode chip that we developed was used for the detection of human chorionic gonadotropin (hCG) sandwich immunoreactivity, as well as gold nanoparticle (AuNP)-conjugated antibodies as labels to facilitate detection. The immune response of the tested specimen and quantitative impedance measurement analysis results were used for identification. An LCR meter, which is a type of electronic test equipment used to measure the inductance (L), capacitance (C), and resistance (R) of an electronic component, was used to detect the electrical signal of the impedance electrode chip, and the changes in immune response were analyzed through impedance value changes to establish the electrical measurement data of hCG. To avoid signal measurement problems caused by poor bonding between the nitrocellulose (NC) membrane and the detection electrode, the NC membrane was fixed directly onto the surface of the detection electrode by a polymer solution phase conversion method, and quantitative impedance signals at different hCG concentrations were successfully measured. The results showed that using this method successfully broke through the traditional naked-eye identification concentration limit of 25 mIU/mL, reaching a lower concentration of 6.25 mIU/ mL, and the coefficient of determination between the impedance value signal and the hCG concentration reached 0.9899, which means that the self-made electrode chip can quantitatively detect and analyze the hCG concentration. [ABSTRACT FROM AUTHOR]

Tuna is an excellent food product, relatively low in calories, that is recommended for a balanced diet. The continuously increasing demand, especially for bluefin-tuna-based food preparations, and its relatively high market price make adulteration by intentionally mixing with other lower-priced tunas more prospective. The development of rapid methods to detect tuna adulteration is a great challenge in food analytical science. We have thus developed a simple, fast, and low-cost molecular rapid test for the visual detection of tuna adulteration. It is the first sensor developed for tuna authenticity testing. The three species studied were Thunnus thynnus (BFT), Thunnus albacares, and Katsuwonus pelamis. DNA was isolated from fresh and heat-treated cooked fish samples followed by PCR. The PCR products were hybridized (10 min) to specific probes and applied to the rapid sensing device. The signal was observed visually in 10–15 min using gold nanoparticle reporters. The method was evaluated employing binary mixtures of PCR products from fresh tissues and mixtures of DNA isolates from heat-treated tissues (canned products) at adulteration percentages of 1–100%. The results showed that the method was reproducible and specific for each tuna species. As low as 1% of tuna adulteration was detected with the naked eye. [ABSTRACT FROM AUTHOR]

Lateral flow immunoassays (LFIA) for antibody detection represent cost-effective and user-friendly tools for serology assessment. This study evaluated a new LFIA prototype developed with a recombinant chimeric antigen from the spike/S and nucleocapsid/N proteins to detect anti-SARS-CoV-2 IgG antibodies. The evaluation of LFIA sensitivity and specificity used 811 serum samples from 349 hospitalized, SARS-CoV-2 RT-qPCR positive COVID-19 patients, collected at different time points and 193 serum samples from healthy controls. The agreement between ELISA results with the S/N chimeric antigen and LFIA results was calculated. The LFIA prototype for SARS-CoV-2 using the chimeric S/N protein demonstrated 85 % sensitivity on the first week post symptoms onset, reaching 94 % in samples collected at the fourth week of disease. The agreement between LFIA and ELISA with the same antigen was 92.7 %, 0.827 kappa Cohen value (95 % CI [0.765–0.889]). Further improvements are needed to standardize the prototype for whole blood use. The inclusion of the novel chimeric S + N antigen in the COVID-19 IgG antibody LFIA demonstrated optimal agreement with results from a comparable ELISA, highlighting the prototype's potential for accurate large-scale serologic assessments in the field in a rapid and user-friendly format.

The performance evaluation of new and old subgrades is critical for the quality and safety of reconstruction and extension projects. It is necessary to achieve rapid and easy performance testing. In this study, a Portable FallingWeight Deflectometer (PFWD) is chosen to rapid evaluate the performance of subgrade. First, a testing area, the reconstruction and expansion project of the Hefei to Dagudian section of the Shanghai-Shaanxi Expressway, is selected. Then, the PFWD modulus Ep of resilient tested by PFWD and the corresponding water content w and compacting degree K tested by the cutting ring method for old subgrade are obtained. And the correlation relationship between Ep and w and K is established. The performance of old subgrade can be rapid obtained by PFWD. Meanwhile, for the new subgrade, the correlation relationship between Ep and bending value L, w and K is established, and the performance can also be rapid tested by PFWD. Finally, a rapid evaluation method for the reconstruction and expansion of subgrade performance was proposed, which aims to provide technical support for ensuring construction quality and safety and provides a technical reference and a theoretical basis for the prediction of similar subgrade performance.

Objectives: To develop a system for the rapid detection of Acinetobacter baumannii 173-p1 antibiotic resistance (to ensure reliable fixation of bacteria on a cantilever without losing their nanomotion, to show that nanomotion is due to bacterial metabolism, to compare the nanomotion of bacteria in suspension form and inside of the biofilms), to study the sensitivity/resistance of A. baumannii 173-p1 to antibiotics (lincomycin, ceftriaxone and doxycycline) using the oscillation method of atomic force microscopy and to evaluate the sensitivity and speed of the method in comparison with the classical disk diffusion method. Methods: The oscillation mode of atomic force microscopy, scanning electron microscopy and the classical disk diffusion method were used for a complex parallel study of A. baumannii 173-p1 antibiotic resistance, which included testing of fixing agents (poly-L-lysine, rosin and fibronectin), comparison of bacterial metabolism in a set of media (normal saline solution, meat-peptone broth and lysogeny broth) and assessment of antibiotic sensitivity/resistance per se. Results: A method for express testing of Acinetobacter baumannii antibiotic resistance using AFM was developed; it is shown that bacterial nanomotion directly correlates with bacteria metabolic activity and that bacterial nanomotion is more easily detected in suspension form, rather than in biofilms. Conclusion: The express testing method gave results that are completely comparable with the classical disk diffusion test and with the results of morphology studies by the SEM method, but it significantly exceeded them in speed, allowing a conclusion to be made on the sensitivity/resistance of bacteria less than an hour after the start of the diagnostics.

Introduction: Zika virus (ZIKV) is a re-emerging flavivirus that poses a significant public health threat. ZIKV exhibits a wide array of non-vector borne human transmission routes, such as sexual transmission, transplacental transmission and blood transfusion. Detection and surveillance of ZIKV is considered paramount in prevention of major outbreaks. With the majority of cases reported in low-resource locations, simple, lowcost detection methods are considered highly desirable. Materials and Methods: Here we have developed a sensitive and specific ZIKV diagnostic using reverse transcription recombinase-aided amplification (RT-RAA) coupled with lateral flow detection (LFD) targeting a highly conserved region of the ZIKV NS1 gene. Results: We show our rapid, isothermal-ZIKV-diagnostic (Iso-ZIKV-Dx) can detect 500 copies of synthetic ZIKV RNA/µL in under 30min at a constant 39°C. Using simulated urine samples, we observed that Iso-ZIKV-Dx also detects as low as 34.28 RNA copies/reaction of ZIKV (MR766 strain). Specificity testing confirmed that our test does not detect any co-circulating flaviviruses (dengue, West Nile, Japanese encephalitis, Murray Valley encephalitis and yellow fever viruses) or chikungunya virus. Sample processing results show complete inactivation of ZIKV (MR766 strain) in 5min at room temperature using our novel viral RNA sample preparation reagent. Furthermore, lateral flow strips testing demonstrates positive diagnoses in as little as 5min in running buffer. Discussion: Contrary to conventional RT-qPCR, our Iso-ZIKV-Dx does not require expensive machinery, specialised laboratory settings or extensively trained personnel. Pre-clinical evaluation demonstrates that our test exhibits robust, in-field capabilities without compromising sensitivity or specificity. When compared to the gold-standard RT-qPCR, our Iso-ZIKV-Dx test offers an array of applications that extend beyond diagnostics alone, including potential for surveillance and monitoring of ZIKV vector competency. [ABSTRACT FROM AUTHOR]

Indonesia began deploying a COVID-19 vaccine in January 2021, prioritising vaccination for high-risk groups such as healthcare workers, the elderly and those with comorbidities, and ending with the general public due to limited vaccine availability. Our study aimed to evaluate antibody response in Indonesians who had received two doses of the vaccine vs. those who had not. The study design was a cohort study involving 46 unvaccinated people and 23 people who had received the second dose of the AstraZeneca vaccine in three months. Methods used for the qualitative and quantitative detection of IgG antibodies included rapid RI-GHA and ELISA tests. Findings showed that positive IgG antibodies qualitatively detected by the rapid RI-GHA test were significantly higher in those vaccinated (60.9%) than in unvaccinated people (26.1%). Using the ELISA assay, all vaccinated individuals qualitatively showed positive antibodies (cut-off ≥4.33 BAU/ml), and the average quantitative titer of anti-SARS-CoV-2 s-RBD IgG was significantly higher in vaccinated (157.06±238.68 BAU/ml) than in unvaccinated (51.90±87.60 BAU/ml) individuals. Some unvaccinated individuals with no history of infection were found to have anti-SARS-CoV-2 antibodies that may have been previously asymptomatic, although their mean antibody titers were certainly lower than those in the 2-dose group. Approximately 56% of vaccinated individuals had antibody titers above 60 BAU/ml as a cut-off for protective threshold, a significantly higher proportion than unvaccinated individuals. In conclusion, vaccination with two doses AstraZeneca increased anti-SARS-CoV-2 antibodies which resulted in enhanced immunity against symptomatic COVID-19. [ABSTRACT FROM AUTHOR]

Purpose: Due to its ability to disseminate worldwide and its multiple resistance trait, Acinetobacter baumannii is becoming a threat for public health worldwide. Cefiderocol (FDC) is a promising broad-spectrum cephalosporin recently approved for treating Gram-negative infection. The aim of this study was to develop a rapid test, namely the rapid FDC Acinetobacter baumannii NP test, for the detection of FDC susceptibility/resistance in A. baumannii since the current FDC susceptibility tests are rather time-consuming (at least 24 h). Materials and methods: The rapid test is based on the reduction of resazurin to resorufin product by bacterial viable cells, thus detecting bacterial growth in the presence of FDC (38.4 mg/L). A color change from blue (resazurin) to violet or pink (resorufin) represents visual detection of bacterial growth. 95 randomly selected A. baumannii isolates were used to evaluate the performance of the rapid FDC Acinetobacter baumannii NP test. Results: The test showed 95.5% (95% CI 78.2–99.2%) and 100.0% (95% CI 95.0–100.0%) of sensitivity and specificity, respectively. All the results were obtained within 4 h30–4 h45 incubation time at 35 °C ± 2 °C, saving virtually one day when compared with currently-used antimicrobial susceptibility tests. The test showed only a single very major error, an isolate with a MIC of 8 mg/L. Conclusions: The rapid FDC Acinetobacter baumannii NP test can be a valuable method which is easier and faster to interpret when compared with the gold standard broth microdilution method. The test showed remarkable performances; hence, it may be suitable for implementation in clinical microbiology routine laboratories. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Inorganic Analytical Chemistry / Zhongguo Wuji Fenxi Huaxue is the property of Beijing Research Institute of Mining & Metallurgy Technology Group and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Pediatría de Atención Primaria is the property of LUA Ediciones 3.0 S.L. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Detection of allergens in foods, including soybean, is relevant for food labelling requirements. Moreover, allergen-specific methods may allow standardisation of allergens in food matrices for use in food challenges as allergy diagnostic approaches. Rapid methods are preferred for screening and along the manufacturing line. Previously, we demonstrated sensitive and specific detection of soybean DNA by combining loop-mediated isothermal amplification (LAMP) and lateral flow device (LFD)-like visualisation. However, lengthy DNA extraction and potential contamination of subsequent by previous LAMP reactions from unclosed LFD may impact its use as a rapid and robust method. Here, we developed a rapid protocol for DNA extraction. Moreover, we identified phenol red for distinct visualisation of positive reactions in permanently closed reaction tubes. The optimised method was validated using complex foods (boiled sausage, instant soup, and chocolate) with known amounts of soybean. Further, its applicability was shown in 12 processed retail foods. Results were verified by orthogonal qPCR. The enhanced LAMP method allowed detection at or below 10 mg soybean per kg processed food. The method provides rapid and easy-to-use screening without the need for detection equipment. Hence, it may serve to verify the presence of soybean ingredients and support a risk-based precautionary labelling of non-ingredient soybean in compound foods. Also, as determination of clinical reaction thresholds before and after allergen immunotherapy (AIT) is both inclusion and exclusion criterion for clinical trials and success parameter of AIT, the method may allow verification of calculable soybean content in provocation meals and thus a standardised administration for threshold determination before and after AIT. [ABSTRACT FROM AUTHOR]

Cyclosporiasis, caused by the coccidian parasite Cyclospora cayetanensis, has emerged as an increasing global public health concern, with the incidence of laboratory-confirmed domestically acquired cases in the US exceeding 10,000 since 2018. A recently published qPCR assay (Mit1C) based on a mitochondrial target gene showed high specificity and good sensitivity for the detection of C. cayetanensis in fresh produce. The present study shows the integration and verification of the same mitochondrial target into a fully automated and streamlined platform that performs DNA isolation, PCR, hybridization, results visualization, and reporting of results to simplify and reduce hands-on time for the detection of this parasite. By using the same primer sets for both the target of interest (i.e., Mit1C) and the internal assay control (IAC), we were able to rapidly migrate the previously developed Mit1C qPCR assay into the more streamlined and automated format Rheonix C. cayetanensisTM Assay. Once the best conditions for detection were optimized and the migration to the fully automated format was completed, we compared the performance of the automated platform against the original "bench top" Mit1C qPCR assay. The automated Rheonix C. cayetanensis Assay achieved equivalent performance characteristics as the original assay, including the same performance for both inclusion and exclusion panels, and it was able to detect as low as 5 C. cayetanensis oocysts in fresh produce while significantly reducing hands-on time. We expect that the streamlined assay can be used as a tool for outbreak and/or surveillance activities to detect the presence of C. cayetanensis in produce samples. [ABSTRACT FROM AUTHOR]

ABSTRACT Rapid HIV tests are critical to HIV surveillance and universal testing and treatment programs. We assessed longitudinal patterns in indeterminate HIV rapid test results in an African population-based cohort. Prospective HIV rapid antibody test results, defined by two parallel rapid tests, among participants aged 15–49 years from three survey rounds of the Rakai Community Cohort Study, Uganda, from 2013 to 2018, were assessed. An indeterminate result was defined as any weak positive result or when one test was negative and the other was positive. A total of 31,405 participants contributed 54,459 person-visits, with 15,713 participants contributing multiple visits and 7,351 participants contributing 3 visits. The prevalence of indeterminate results was 2.7% (1,490/54,469). Of the participants with multiple visits who initially tested indeterminate (n = 591), 40.4% were negative, 18.6% were positive, and 41.0% were indeterminate at the subsequent visit. Of the participants with two consecutive indeterminate results who had a third visit (n = 67), 20.9% were negative, 9.0% were positive, and 70.2% remained indeterminate. Compared to a prior negative result, a prior indeterminate result was strongly associated with a subsequent indeterminate result [adjusted prevalence ratio, 23.0 (95% CI = 20.0–26.5)]. Compared to men, women were more likely to test indeterminate than negative [adjusted odds ratio, 2.3 (95% CI = 2.0–2.6)]. Indeterminate rapid HIV test results are highly correlated within an individual and 0.6% of the population persistently tested indeterminate over the study period. A substantial fraction of people with an indeterminate result subsequently tested HIV positive at the next visit, underscoring the importance of follow-up HIV testing protocols.IMPORTANCERapid HIV tests are a critical tool for expanding HIV testing and treatment to end the HIV epidemic. The interpretation and management of indeterminate rapid HIV test results pose a unique challenge for connecting all people living with HIV to the necessary care and treatment. Indeterminate rapid HIV test results are characterized by any weak positive result or discordant results (when one test is negative and the other is positive). We systematically tested all participants of a Ugandan population-based, longitudinal cohort study regardless of prior test results or HIV status to quantify longitudinal patterns in rapid HIV test results. We found that a substantial fraction (>15%) of participants with indeterminate rapid test results subsequently tested positive upon follow-up testing at the next visit. Our findings demonstrate the importance of follow-up HIV testing protocols for indeterminate rapid HIV test results.

The COVID-19 pandemic has primarily been controlled by testing for SARS-CoV-2 infections. Despite vaccines, testing will remain crucial for surveillance and screening, allowing for the detection of new variants in a timely manner and to isolate the infected people to lower the danger of the disease spreading further. The research study attempts to found out the efficiency of Reverse-transcriptase polymerase chain reaction (RT-PCR) and Rapid antigen tests in symptomatic COVID-19 patients at tertiary care hospitals. The research was performed on 1000 patients, both In-patients and Out-patients, who presented with COVID-19 symptoms. SARS-COV-2 nucleocapsid protein antigen was detected qualitatively with rapid antigen test in human nasal specimens through the immuno-chromatographic assay. The rapid test results were compared with a molecular test RT-PCR in which FAM, HEX, and ROX were the indicator dyes for the RdRp gene, E gene, and the internal control (RNAse P), respectively. Nearly 322 cases were positive with both RT-PCR and rapid antigen test methods. Fifty-nine samples yielded negative results with the rapid antigen test and positive with PCR. Three samples were negative with RT-PCR and positive with the rapid antigen test. The findings from our study show that the common symptoms are fever 92.2% and cough 74.1% in the reported test population. But in confirmed cases of RT-PCR showed cough at 74.1% was more prevalent, followed by fever at 41.3%. Rapid antigen test showed a overall sensitivity and specificity of 85.3% and 99.5%. According to World Health Organization, rapid antigen detection tests meet the minimum performance requirements of ≥80% sensitivity and ≥97% specificity. Hence, the present study meets this criterion and may perhaps be a probable tool for point-of-care in hospital settings.

Background: Throat swab cultures still remain the gold standard for the confirmation of Group A Streptococcal (GAS) pharyngitis but Rapid Antigen detection Test (RADT) are increasingly becoming popular. Recent studies show RADT can be as sensitive and specific as the throat swab culture. This study aimed to determine the utility of RADT in the diagnosis of GAS pharyngitis in children at the Federal Teaching Hospital Gombe. Methods: A cross-sectional study that included 324 children aged 3- 18 years presenting with sorethroat at the out-patient department, consecutively recruited between April and September 2018. Socio-demographic and clinical findings were document. Throat swab samples were taken for RADT using Encode strep A Rapid antigen test and culture on 5% sheep blood agar. The sensitivity, specificity and positive and negative predictive values of the RADT was determined against throat swab cultures, which is the gold standard. Results: There were 190 (58.6%) females and Male to Female ratio of 1:1.4. The mean age was 8.3± 3.9 years. Only 125 (38.6%) of the participants were from low social class and 162 (62.3%) are from overcrowded households. Sociodemographic factors have no effect on the performance of the RADT, however, the use of antibiotics within 2 weeks prior to presentation significantly affected the performance of the RADT 1.4% (1/64) compared to 28.1% (73/260) for those without prior antibiotics use p

Vibrio qinghaiensis sp. Q67 was used as an indicator strain to study the toxic effects of five common pesticides, pirimicarb (PIR), carbaryl (CAR), glyphosate (GLY), dimethoate (DIM), and acephate (ACE). The toxicities were evaluated by toxic unit and concentration addition model. The results showed that the toxicity of the pesticides decreased in the following order: PIR > GLY > CAR > DIM > ACE. Four of the 10 binary mixtures of these five pesticides showed antagonistic toxic effects, while the remaining six showed additive or synergistic toxic effects. Among the seven five-component mixtures designed by uniform design ray, four showed synergistic and then antagonistic effects, while the remaining three showed a consistent antagonistic effect. The toxicity of the five-component mixtures was strongly positively correlated with the concentration of CAR (r = 0.947). Based on this result, a linear model was established to predict the toxicity of pesticide mixtures. This method has the advantages of low cost, rapidity, simplicity and convenience, which can be used as a potential preliminary screening method for rapid detection of pesticide residues in agricultural products and foods.

Vector surveillance of Plasmodium falciparum is critical for monitoring and reducing one of the most severe forms of malaria, which causes high morbidity and mortality in children under five and pregnant women. Here we developed a rapid and highly sensitive test for the detection of P. falciparum (Pf)-infected mosquitoes (Rapid Pf test), with high suitability for low-resource vector surveillance implementation. The Rapid Pf test had similar analytical sensitivity to laboratory-based tests, detecting down to 4 copies/μL of a 18S rRNA DNA standard. In addition, the Rapid Pf test could be completed in less than 30 minutes, and only required a liquid sample preparation reagent, pestle, tube, and 39°C heating block for operation, indicating amenability for low-resource implementation. Diagnostic testing was performed using Anopheles stephensi mosquitoes, either uninfected, or fed with P. falciparum gametocyte cultures. These P. falciparum fed mosquitoes were determined to have 79% infection prevalence based on parallel microscopy and qPCR testing on a subset of 19 mosquitoes. However, our Rapid Pf test determined a 90% positive test rate when testing individual infected mosquitoes (n=30), and did not detect 40 uninfected mosquitoes regardless of blood-fed status (n=40), suggesting the true prevalence of infection in the mosquitoes may have been higher than calculated by qPCR and microscopy. The Rapid Pf test was demonstrated to detect infection in individual mosquitoes (both fresh and frozen/thawed), as well as pools of 1 infected mosquito mixed with 19 known uninfected mosquitoes, and individual mosquitoes left in traps for up to 8 days. After testing on infected and uninfected mosquitoes (n=148) the Rapid Pf test was conservatively estimated to achieve 100% diagnostic sensitivity (95% confidence interval, CI: 91%-100%) and 97% diagnostic specificity (CI: 92%-99%) compared to the estimated prevalence from combined microscopy and qPCR results. These results indicate the Rapid Pf test could provide a highly effective tool for weekly surveillance of infected mosquitoes, to assist with P. falciparum monitoring and intervention studies.

Indonesia began deploying a COVID‑19 vaccine in January 2021, prioritising vaccination for high‑risk groups such as healthcare workers, the elderly and those with comorbidi‑ ties, and ending with the general public due to limited vaccine availability. Our study aimed to evaluate antibody response in Indonesians who had received two doses of the vaccine vs. those who had not. The study design was a cohort study involving 46 unvaccinated people and 23 people who had received the second dose of the AstraZeneca vaccine in three months. Methods used for the qualitative and quantitative detection of IgG antibodies included rapid RI‑GHA and ELISA tests. Findings showed that positive IgG antibodies qualitatively detected by the rapid RI‑GHA test were significantly higher in those vaccinated (60.9%) than in unvaccinated people (26.1%). Using the ELISA assay, all vaccinated individuals qualitatively showed positive antibodies (cut‑off ≥4.33 BAU/ml), and the average quantitative titer of anti‑SARS‑CoV‑2 s‑RBD IgG was significantly higher in vaccinated (157.06±238.68 BAU/ml) than in unvaccinated (51.90±87.60 BAU/ml) individuals. Some unvaccinated individuals with no history of infection were found to have anti‑SARS‑CoV‑2 antibodies that may have been previously asymptomatic, although their mean antibody titers were certainly lower than those in the 2‑dose group. Approximately 56% of vaccinated individuals had antibody titers above 60 BAU/ml as a cut‑off for protective threshold, a significantly higher proportion than unvaccinated individuals. In conclusion, vaccination with two doses AstraZeneca increased anti‑SARS‑CoV‑2 antibodies which resulted in enhanced immunity against symptomatic COVID‑19.

Abstract: Melon is one of the most consumed cucurbits and commercially significant cucurbits in the world. It is planted predominantly through seedlings, and seed quality is essential in this process. Among the rapid tests used in seed analysis, the pH testing of exudate has shown promise for several cultivated species. Thus, this study aimed to verify the viability of melon (Cucumis melo L.) seeds through the pH testing of exudate. For this, seeds of the hybrids Premier and Supreme (three lots each) were subjected to germination and vigor tests (first germination count, tetrazolium, accelerated aging, emergence speed index, and emergence). The pH testing of exudate was evaluated using phenolphthalein and sodium carbonate solutions. The results indicated the possibility of classifying the lots into three quality levels for both hybrids, similar to most tests used to assess initial quality. Therefore, the pH testing of exudate was effective in determining the viability of lots of melon seeds and can be conducted with a soaking period of 120 minutes at 25 or 30 °C.

IntroductionZika virus (ZIKV) is a re-emerging flavivirus that poses a significant public health threat. ZIKV exhibits a wide array of non-vector borne human transmission routes, such as sexual transmission, transplacental transmission and blood transfusion. Detection and surveillance of ZIKV is considered paramount in prevention of major outbreaks. With the majority of cases reported in low-resource locations, simple, low-cost detection methods are considered highly desirable.Materials and MethodsHere we have developed a sensitive and specific ZIKV diagnostic using reverse transcription recombinase-aided amplification (RT-RAA) coupled with lateral flow detection (LFD) targeting a highly conserved region of the ZIKV NS1 gene.ResultsWe show our rapid, isothermal-ZIKV-diagnostic (Iso-ZIKV-Dx) can detect 500 copies of synthetic ZIKV RNA/μL in under 30 min at a constant 39°C. Using simulated urine samples, we observed that Iso-ZIKV-Dx also detects as low as 34.28 RNA copies/reaction of ZIKV (MR766 strain). Specificity testing confirmed that our test does not detect any co-circulating flaviviruses (dengue, West Nile, Japanese encephalitis, Murray Valley encephalitis and yellow fever viruses) or chikungunya virus. Sample processing results show complete inactivation of ZIKV (MR766 strain) in 5 min at room temperature using our novel viral RNA sample preparation reagent. Furthermore, lateral flow strips testing demonstrates positive diagnoses in as little as 5 min in running buffer.DiscussionContrary to conventional RT-qPCR, our Iso-ZIKV-Dx does not require expensive machinery, specialised laboratory settings or extensively trained personnel. Pre-clinical evaluation demonstrates that our test exhibits robust, in-field capabilities without compromising sensitivity or specificity. When compared to the gold-standard RT-qPCR, our Iso-ZIKV-Dx test offers an array of applications that extend beyond diagnostics alone, including potential for surveillance and monitoring of ZIKV vector competency.

The COVID-19 pandemic has primarily been controlled by testing for SARS-CoV-2 infections. Despite vaccines, testing will remain crucial for surveillance and screening, allowing for the detection of new variants in a timely manner and to isolate the infected people to lower the danger of the disease spreading further. The research study attempts to found out the efficiency of Reverse-transcriptase polymerase chain reaction (RT-PCR) and Rapid antigen tests in symptomatic COVID-19 patients at tertiary care hospitals. The research was performed on 1000 patients, both In-patients and Out-patients, who presented with COVID-19 symptoms. SARS-COV-2 nucleocapsid protein antigen was detected qualitatively with rapid antigen test in human nasal specimens through the immuno-chromatographic assay. The rapid test results were compared with a molecular test RT-PCR in which FAM, HEX, and ROX were the indicator dyes for the RdRp gene, E gene, and the internal control (RNAse P), respectively. Nearly 322 cases were positive with both RT-PCR and rapid antigen test methods. Fifty-nine samples yielded negative results with the rapid antigen test and positive with PCR. Three samples were negative with RT-PCR and positive with the rapid antigen test. The findings from our study show that the common symptoms are fever 92.2% and cough 74.1% in the reported test population. But in confirmed cases of RT-PCR showed cough at 74.1% was more prevalent, followed by fever at 41.3%. Rapid antigen test showed a overall sensitivity and specificity of 85.3% and 99.5%. According to World Health Organization, rapid antigen detection tests meet the minimum performance requirements of ≥80% sensitivity and ≥97% specificity. Hence, the present study meets this criterion and may perhaps be a probable tool for point-of-care in hospital settings. [ABSTRACT FROM AUTHOR]

Self-administered HIV testing may be a promising strategy to improve testing in hard-to-reach young adults, provided they are aware of and willing to use oral HIV self-testing (HIVST). This study examined awareness of and willingness to use oral HIVST among 350 high-risk young adults, aged 18–22, living in Kenya's informal urban settlements. Bivariate and multivariate logistic regressions were used to examine differences in HIVST awareness and willingness by demographic and sexual risk factors. Findings showed that most participants were male (56%) and less than 20 years old (60%). Awareness of oral HIVST was low (19%). However, most participants (75%) were willing to use an oral HIV self-test in the future and ask their sex partner(s) to self-test before having sex (77%). Women (OR = 1.80, 95%CI:1.11, 2.92), older participants (aged 20+) (OR = 2.57, 95% CI:1.48, 4.46), and more educated participants (OR = 2.25, 95%CI:1.36, 3.70) were more willing to use HIVST as compared to men, teen-aged, and less educated participants, respectively. Young adults who reported recent engagement in high-risk sexual behaviors, such as unprotected sex, sex while high or drunk, or sex exchange, were significantly less likely to be willing to use an oral HIV self-test kit (OR = 0.34, 95%CI:0.13,0.86). Those with the highest monthly income (OR = 0.47, 95%CI: 0.25, 0.89) were also less willing to use HIVST. More community- and peer-based efforts are needed to highlight the range of benefits of HIVST (i.e., social, clinical, and structural) to appeal to various youth demographics, in addition to addressing concerns relating to HIVST. [ABSTRACT FROM AUTHOR]

Respiratory infections constitute a major reason for infants and children seeking medical advice and visiting health facilities, thus remaining a significant public threat with high morbidity and mortality. The predominant viruses causing viral respiratory infections are influenza A and B viruses (Flu-A, Flu-B), respiratory syncytial virus (RSV), adenovirus and coronaviruses. We aimed to record the proportion of RSV, SARS-CoV-2, influenza A/B and adenovirus cases with rapid antigen tests and validate the results with RT-PCR assays of upper respiratory specimens with a wide range of viral loads and (co)-infection patterns in children. Clinical samples were collected from early symptomatic children (presenting with fever and/or cough and/or headache within 5–7 days). The surveillance program was conducted in five private pediatric dispensaries and one pediatric care unit, from 10 January 2023 to 30 March 2023 in central Greece. The total sample of specimens collected was 784 young children and infants, of which 383 (48.8%) were female and 401 were male (51.2%). The mean age of participants was 7.3 + 5.5 years. The sensitivity of the FLU A & B test was 91.15% (95% CI: 84.33–95.67%), and the specificity was 98.96% (95% CI: 97.86–99.58%). The sensitivity and specificity of the adenovirus and RSV test was {92.45% (95% CI: 81.79–97.91%), 99.32% (95% CI: 98.41–99.78%)} and {92.59% (95% CI: 75.71–99.09%), 99.47% (95% CI: 98.65–99.86%)} respectively. Lastly, the sensitivity of the SARS-CoV-2 test was 100.00% (95% CI: 79.41–100.00%) and the specificity was 99.74% (95% CI: 99.06–99.97%). We recorded a proportion of 14.3% and 3.44% for influenza A and B, respectively, followed by a proportion of 6.9% for adenovirus, a proportion of 3.7% for RSV, and finally, a proportion of 2.3% for SARS-CoV-2. The combination of a new multiple rapid test with multiple antigens will probably be a useful tool with a financial impact for health systems targeting the early detection and appropriate treatment of respiratory infections in emergency departments in primary health care facilities. [ABSTRACT FROM AUTHOR]

The dramatic rise of global incidence of dengue infection has generated an urgent yet an unmet need for an accurate and validated diagnostic test for DENV infection. A total of 81 serum samples were enlisted to dengue NS1 ICT test and dengue NS1 ELISA test and oxidative stress markers including NO, MDA and SOD activity were measured. Out of 81 samples, 41 were found positive by ELISA (DENV group; n=41) whereas 28 were found positive by ICT test. Out of 81 samples; 40 were found negative (control group; n=40) by ELISA whereas 53 were found negative by ICT test. There were 28 samples which were found positive by both ICT and ELISA test. There were 40 samples which were found negative by both ICT and ELISA. The sensitivity and specificity of ICT test were 68.29% and 100% when compared with ELISA. The positive cases of dengue was confirmed by platelet count having significantly higher level of platelet counts (P [ J Adv Biotechnol Exp Ther 2023; 6(1.000): 94-108]

African swine fever (ASF) is a contagious disease of wild boar and domestic pigs notifiable to the World Organisation for Animal Health due to its high socio-economic impact. ASF is caused by the complex ASF virus (ASFV), and it can present different clinical manifestations that can be confused with other diseases; for this reason, laboratory testing is necessary for the proper diagnosis of clinically suspected animals. Despite the efforts put into it over decades, no treatment or safe vaccine is globally available, and disease control is based on early diagnosis and the implementation of strict biosecurity measures. In this context, rapid tests have the potential to accelerate and facilitate the identification of infected animals by giving fast on-site results. In this work, we improved the available point-of-care assays for the diagnosis of the disease by the development of a more specific antigen test and a more sensitive antibody test. This antibody detection test allowed for the earlier detection of infected animals than two commercial indirect ELISAs (statistically significant). Moreover, we developed a combined dual rapid test, unifying, in the same cassette, an antigen detection strip and an antibody detection strip. In this study, we confirmed that this combo approach is a useful tool for implementing rapid tests in the field since it increases the percentage of positive samples detected, even when PCR turns negative, while maintaining a good specificity.

Background & objectives: High transmissibility of the SARS-CoV-2 has significant implications on healthcare workers' safety, preservation, handling, transportation and disposal of the deceased bodies. The objective of this study was to detect SARS-CoV-2 antigen in nasopharyngeal samples and its implications in handling and care of COVID-19 deceased bodies. Methods: A study was conducted at a dedicated COVID-19 centre on deceased individuals from April to December 2020. Rapid antigen test (RAT) and reverse transcription (RT)-PCR was compared on all the SARS-CoV-2 positive cadavers recruited in the study. Results: A total of 115 deceased individuals were included in the study. Of these, 79 (68.7%) were male and 36 (31.3%) were female and majority were in the age group of 51-60 yr [31 (27%)]. SARS-CoV-2 antigen test was positive in 32 (27.8%) and negative in 83 (72.1%) individuals. The mean time interval between deaths to the sample collection was 13.2 h with interquartile range of eight to 20 h. Reverse transcription (RT)-PCR was used as the reference test and 24 (20.9%) cases were true positive; 93.6 per cent [95% confidence interval (CI) 88.8-98.4%] sensitivity, 45.2 per cent (95% CI 35.5-55%) specificity, 60.2 per cent (95% CI 50.6-69.8%) positive predictive value and 88.8 per cent (95% CI 82.7-95%) negative predictive value of antigen test was computed. Interpretation & conclusions: SARS-CoV-2 antigen test was positive beyond 19 h in COVID-19 deceased individuals. Antigen test was found to be highly sensitive in the deceased. Patients, suspected of having died due to COVID-19, can be screened by this method. As infectiousness of the virus in the deceased bodies cannot be directly concluded from either the antigen or RT-PCR test, yet possible transmission cannot be completely ruled out. Strict infection control measures need to be followed during the handling and clearance of COVID-19 cadavers. [ABSTRACT FROM AUTHOR]

Introduction: In most of the endemic areas, the detection of CL is based on searching for amastigotes using the direct smear method. Since expert microscopists are not usually available in every laboratory, false diagnoses are a disaster that happens. Therefore, the aim of current research is to evaluate the validity of the CL Detect™ Rapid Test (CDRT) for diagnosis CL in comparison to direct smear and polymerase chain reaction (PCR) methods. Methods: A total of 70 patients with skin lesions suspected to be CL were recruited. Skin samples from the lesions were collected and used for direct microscopic examination and the PCR method. Furthermore, the skin sample was collected in accordance with the manufacturer's instructions for the CDRT-based rapid diagnostic test. Results: Of 70 samples, 51 and 35 samples were positive by direct smear examination and the CDRT, respectively. The PCR showed positive results in 59 samples; 50 and 9 samples were identified as Leishmania major and Leishmania tropica, respectively. The sensitivity and specificity were calculated to be 68.6% (95% CI 54.11–80.89%) and 100% (95% CI 82.35–100%). When the results of CDRT were compared to the microscopic examinations, an agreement of 77.14% was seen between the CDRT and microscopic examination. In addition, the sensitivity and specificity were 59.32% (95% CI 45.75–71.93%) and 100% (95% CI 71.5–100%) when the CDRT was compared to PCR assay (as gold standard) and an agreement (65.71%) was found between CDRT and PCR assay. Conclusion: As the CDRT is simple, rapid, and does not require great proficiency, it is recommended for use in the detection of CL caused by L. major or L. tropica as a diagnostic method, especially in areas with limited access to expert microscopists. [ABSTRACT FROM AUTHOR]

Background: Toxoplasmosis is a protozoan disease caused by Toxoplasma gondii. Different T. gondii confirmatory techniques, including serologic methods, are available to detect the presence of the parasite. Among serology techniques, immunochromatographic rapid testing could be a reliable alternative to serologic laboratory techniques. Objective: This study evaluated a commercial immunochromatographic test (FASTest TOXOPLASMA g) in seronegative and seropositive cats. Methods: Two indirect immunofluorescence antibody reference tests, an in‐house technique, and a commercial test were used to classify 292 feline serum samples. The rapid test was evaluated in different groups of cats, including healthy seronegative cats (n = 121), seropositive cats with variable anti‐Toxoplasma antibodies (n = 146), and cats with positive serologic results for other pathogens (n = 25). The sensitivity, specificity, accuracy, receiver operating characteristic curves, and kappa statistics were analyzed as performance measures. Results: Of the 292 samples, 146 were classified as T. gondii seropositive and 146 as T. gondii seronegative. Concordant results were obtained for all samples using immunofluorescence antibody tests. The diagnostic measures of this rapid test showed 98.63% sensitivity and 100% specificity, and 99.32% accuracy. The kappa statistics value was 0.986, and the area under the receiver operating characteristic curve was 0.993. Conclusions: This rapid test showed diagnostic measurements similar to those of traditional quantitative serologic methods. In situations where laboratory techniques are not available, this test, under clinical conditions, could be a useful alternative to obtain accurate results rapidly. [ABSTRACT FROM AUTHOR]

The clinical impact of rapid sample-to-answer "syndromic" multiplex polymerase chain reaction (PCR) testing for respiratory viruses is not clearly established. We performed a systematic literature review and meta-analysis to evaluate this impact for patients with possible acute respiratory tract infection in the hospital setting. We searched EMBASE, MEDLINE, and Cochrane databases from 2012 to present and conference proceedings from 2021 for studies comparing clinical impact outcomes between multiplex PCR testing and standard testing. Twenty-seven studies with 17,321 patient encounters were included in this review. Rapid multiplex PCR testing was associated with a reduction of − 24.22 h (95% CI −28.70 to −19.74 h) in the time to results. Hospital length of stay was decreased by −0.82 days (95% CI −1.52 to −0.11 days). Among influenza positive patients, antivirals were more likely to be given (RR 1.25, 95% CI 1.06–1.48) and appropriate infection control facility use was more common with rapid multiplex PCR testing (RR 1.55, 95% CI 1.16–2.07). Our systematic review and meta-analysis demonstrates a reduction in time to results and length of stay for patients overall along with improvements in appropriate antiviral and infection control management among influenza-positive patients. This evidence supports the routine use of rapid sample-to-answer multiplex PCR testing for respiratory viruses in the hospital setting. • The clinical impact of rapid multiplex testing for respiratory viruses is not known. • This meta-analysis of 27 studies evaluates the impact compared with standard testing. • Rapid multiplex testing was associated with reduced time to results and length of stay. • Rapid multiplex testing improved antiviral use and infection control in influenza patients. • These results support rapid multiplex PCR tests in suspected respiratory infections. [ABSTRACT FROM AUTHOR]