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Objectives: To describe the in vivo emergence of ceftazidime-avibactam resistance in GES-type carbapenemases and to characterize an unusual outbreak of GES-6-producing Serratia marcescens during the COVID-19 pandemic in Spain., Methods: Retrospective study to describe a GES-CPSM outbreak based on whole genome sequencing and antimicrobial susceptibility testing (AST). Transferability of bla GES -carrying plasmid was assessed by conjugation experiments., Results: In December 2020, we identified a cluster of S. marcescens harbouring bla GES-6 involving 9 patients. Whole-genome sequence analysis revealed a clonal relationship (≤3 SNPs) between the first isolates identified in each of the evolved patients and environmental samples with GES-CPSM detection. Plasmid analysis showed that the bla GES-6 gene was located in an IncQ3-type plasmid. Triparental mating experiments using a helper plasmid demonstrated mobilization of the bla GES-6 -carrying plasmid. Our results also demonstrate within-host evolution in S. marcescens isolates, leading to a transition from bla GES-6 to the new bla GES-55 , caused by the P162S mutation, in a subsequent infection in one of the affected patients. In bla GES-55 we identified emergence of ceftazidime-avibactam resistance along with an increase of carbapenems susceptibility. This patient had been treated with a 14-day course of ceftazidime-avibactam. AST of the transformants bearing bla GES-6 and bla GES-55 plasmids, confirmed susceptibility variation affecting ceftazidime-avibactam and carbapenems., Conclusions: We report an unusual outbreak of GES-6 whose incidence is becoming increasing. Transition from GES-6 to GES-55 may readily occur in vivo leading to ceftazidime-avibactam resistance, which brings to the fore the critical need for developing more accurate diagnosis tools for detection of GES β-lactamases and optimise the use of antimicrobials., (Copyright © 2024 Elsevier Ltd and International Society of Antimicrobial Chemotherapy. All rights reserved.)

Background: Community-acquired (CA) and healthcare-associated (HCA) infections caused by carbapenemase-producing Enterobacterales (CPE) are not well characterized. The objective was to provide detailed information about the clinical and molecular epidemiological features of nosocomial, HCA and CA infections caused by carbapenemase-producing Klebsiella pneumoniae (CP-Kp) and Escherichia coli (CP-Ec)., Methods: A prospective cohort study was performed in 59 Spanish hospitals from February to March 2019, including the first 10 consecutive patients from whom CP-Kp or CP-Ec were isolated. Patients were stratified according to acquisition type. A multivariate analysis was performed to identify the impact of acquisition type in 30-day mortality., Results: Overall, 386 patients were included (363 [94%] with CP-Kp and 23 [6%] CP-Ec); in 296 patients (76.3%), the CPE was causing an infection. Acquisition was CA in 31 (8.0%) patients, HCA in 183 (47.4%) and nosocomial in 172 (48.3%). Among patients with a HCA acquisition, 100 (54.6%) had been previously admitted to hospital and 71 (38.8%) were nursing home residents. Urinary tract infections accounted for 19/23 (82.6%), 89/130 (68.5%) and 42/143 (29.4%) of CA, HCA and nosocomial infections, respectively. Overall, 68 infections (23%) were bacteremia (8.7%, 17.7% and 30.1% of CA, HCA and nosocomial, respectively). Mortality in infections was 28% (13%, 14.6% and 42.7% of CA, HCA and nosocomial, respectively). Nosocomial bloodstream infections were associated with increased odds for mortality (adjusted OR, 4.00; 95%CI 1.21-13.19)., Conclusions: HCA and CA infections caused by CPE are frequent and clinically significant. This information may be useful for a better understanding of the epidemiology of CPE., (© 2024. The Author(s).)

Background: Extended-spectrum ß-lactamase-producing Enterobacterales (ESBL-E) are a serious threat among emerging antibiotic resistant bacteria. Particularly, the number of cases of ESBL-E infections reported in children has been increasing in recent years, and approved antibiotic treatments for this age group are limited. However, information regarding the prevalence of colonization in European children, risk factors associated with colonization, and the characteristics of the colonizing strains is scarce. The aims of this study were to determine the prevalence of ESBL-E colonization in fecal samples of apparently healthy schoolchildren, to identify lifestyle routines associated with colonization, and to characterize clonal relationships and mechanisms of resistance in ESBL-E isolates., Methods: A cohort of 887 healthy children (3-13  years old) from seven primary and secondary schools in the Madrid metropolitan area was recruited between April-June 2018, and sociodemographic information and daily habits were collected. Fecal samples were screened for ESBL-E carriage in selective medium. ESBL-E isolates were further characterized by assessing molecular epidemiology (PFGE and MLST), ESBL gene carriage, and antibiotic resistance profile. This information was analyzed in conjunction with the metadata of the participants in order to identify external factors associated with ESBL-E carriage., Results: Twenty four ESBL-E, all but one Escherichia coli, were detected in 23 children (prevalence: 2.6%; 95% CI: 1.6-3.6%). Of these, seven contained the bla CTX-M-14 allele, five the bla CTX-M-15 , five the bla SHV-12 , three the bla CTX-M-27 , three the bla CTX-M-32 , and one the bla CTX-M-9 . Significant clonal diversity was observed among the isolates that grouped into 22 distinct clusters (at <85% similarity of PFGE profile). ESBL-producing E. coli isolates belonged to 12 different STs, with ST10 (25%) and ST131 (17%) being the most frequent. Apart from ß-lactams, resistance to trimethoprim/sulfamethoxazole (46%), ciprofloxacin (33%), levofloxacin (33%), tobramycin (21%), and gentamicin (8%) were the most frequently detected., Conclusion: The prevalence of ESBL-E in the studied cohort of children was lower than the average colonization rate previously detected in Europe for both children and adults. E. coli was the main ESBL-producing species detected and CTX-M were the most frequently identified ESBLs. High ST diversity suggests polyclonal dissemination. Compared to other STs, ST131 isolates were associated with resistance to various antimicrobials., Competing Interests: MM was founder and stockholder of the biotechnology spin-off company Vaxdyn, which develops vaccines for infections caused by MDR bacteria. Vaxdyn had no role in the elaboration of this manuscript. The remaining 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 © 2023 López-Siles, Moure, Muadica, Sánchez, Cruces, Ávila, Lara, Köster, Dashti, Oteo-Iglesias, Carmena and McConnell.)

The high demand for COVID-19 diagnosis overwhelmed reference hospitals. Regional laboratories had to incorporate molecular technology to respond to the emergency. This work described the implementation of molecular diagnostic tools and the detection of SARS-CoV-2, in a regional hospital with no previous experience, from October 2020 to March 2022. The laboratory structure was significantly modified. The staff grew from 3 to 4 clinical microbiologists, and from 7 to 17 laboratory technicians to provide 24/7 coverage. A total of 144,442 samples were processed during the period of study. The highest peaks were reached in July 2021 with 25,285 samples processed, and between December 2021 and January 2022, with 32,245. COVID-19 pandemic has represented not only the challenge, but the opportunity to introduce Nucleic Acid Amplification Techniques (NAAT) in inexperienced laboratories. These secondary settings have shown an extraordinary ability to adapt and cannot be left behind in the progress of diagnostic techniques.

Trypanosoma cruzi infection has expanded globally through human migration. In Spain, the mother-to-child route is the mode of transmission contributing to autochthonous Chagas disease (CD); however, most people acquired the infection in their country of origin and were diagnosed in the chronic phase (imported chronic CD). In this context, we assessed the quantitative potential of the Loopamp Trypanosoma cruzi detection kit (Sat-TcLAMP) based on satellite DNA (Sat-DNA) to determine parasitemia levels compared to those detected by real-time quantitative PCRs (qPCRs) targeting Sat-DNA (Sat-qPCR) and kinetoplast DNA minicircles (kDNA-qPCR). This study included 173 specimens from 39 autochthonous congenital and 116 imported chronic CD cases diagnosed in Spain. kDNA-qPCR showed higher sensitivity than Sat-qPCR and Sat-TcLAMP. According to all quantitative approaches, parasitemia levels were significantly higher in congenital infection than in chronic CD (1 × 10 -1 to 5 × 10 5 versus >1 × 10 -1 to 6 × 10 3 parasite equivalents/mL, respectively [ P  < 0.001]). Sat-TcLAMP, Sat-qPCR, and kDNA-qPCR results were equivalent at high levels of parasitemia ( P  = 0.381). Discrepancies were significant for low levels of parasitemia and older individuals. Differences between Sat-TcLAMP and Sat-qPCR were not qualitatively significant, but estimations of parasitemia using Sat-TcLAMP were closer to those by kDNA-qPCR. Parasitemia changes were assessed in 6 individual cases in follow-up, in which trends showed similar patterns by all quantitative approaches. At high levels of parasitemia, Sat-TcLAMP, Sat-qPCR, and kDNA-qPCR worked similarly, but significant differences were found for the low levels characteristic of late chronic CD. A suitable harmonization strategy needs to be developed for low-level parasitemia detection using Sat-DNA- and kDNA-based tests. IMPORTANCE Currently, molecular equipment has been introduced into many health care centers, even in low-income countries. PCR, qPCR, and loop-mediated isothermal amplification (LAMP) are becoming more accessible for the diagnosis of neglected infectious diseases. Chagas disease (CD) is spreading worldwide, and in countries where the disease is not endemic, such as Spain, the parasite Trypanosoma cruzi is transmitted from mother to child (congenital CD). Here, we explore why LAMP, aimed at detecting T. cruzi parasite DNA, is a reliable option for the diagnosis of congenital CD and the early detection of reactivation in chronic infection. When the parasite load is high, LAMP is equivalent to any qPCR. In addition, the estimations of T. cruzi parasitemia in patients living in Spain, a country where the disease is not endemic, resemble natural evolution in areas of endemicity. If molecular tests are introduced into the diagnostic algorithm for congenital infection, early diagnosis and timely treatment would be accomplished, so the interruption of vertical transmission can be an achievable goal.

Objectives: CARB-ES-19 is a comprehensive, multicenter, nationwide study integrating whole-genome sequencing (WGS) in the surveillance of carbapenemase-producing K. pneumoniae (CP-Kpn) and E. coli (CP-Eco) to determine their incidence, geographical distribution, phylogeny, and resistance mechanisms in Spain., Methods: In total, 71 hospitals, representing all 50 Spanish provinces, collected the first 10 isolates per hospital (February to May 2019); CPE isolates were first identified according to EUCAST (meropenem MIC > 0.12 mg/L with immunochromatography, colorimetric tests, carbapenem inactivation, or carbapenem hydrolysis with MALDI-TOF). Prevalence and incidence were calculated according to population denominators. Antibiotic susceptibility testing was performed using the microdilution method (EUCAST). All 403 isolates collected were sequenced for high-resolution single-nucleotide polymorphism (SNP) typing, core genome multilocus sequence typing (cgMLST), and resistome analysis., Results: In total, 377 (93.5%) CP-Kpn and 26 (6.5%) CP-Eco isolates were collected from 62 (87.3%) hospitals in 46 (92%) provinces. CP-Kpn was more prevalent in the blood (5.8%, 50/853) than in the urine (1.4%, 201/14,464). The cumulative incidence for both CP-Kpn and CP-Eco was 0.05 per 100 admitted patients. The main carbapenemase genes identified in CP-Kpn were bla OXA-48 (263/377), bla KPC-3 (62/377), bla VIM-1 (28/377), and bla NDM-1 (12/377). All isolates were susceptible to at least two antibiotics. Interregional dissemination of eight high-risk CP-Kpn clones was detected, mainly ST307/OXA-48 (16.4%), ST11/OXA-48 (16.4%), and ST512-ST258/KPC (13.8%). ST512/KPC and ST15/OXA-48 were the most frequent bacteremia-causative clones. The average number of acquired resistance genes was higher in CP-Kpn (7.9) than in CP-Eco (5.5)., Conclusion: This study serves as a first step toward WGS integration in the surveillance of carbapenemase-producing Enterobacterales in Spain. We detected important epidemiological changes, including increased CP-Kpn and CP-Eco prevalence and incidence compared to previous studies, wide interregional dissemination, and increased dissemination of high-risk clones, such as ST307/OXA-48 and ST512/KPC-3., 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 © 2022 Cañada-García, Moure, Sola-Campoy, Delgado-Valverde, Cano, Gijón, González, Gracia-Ahufinger, Larrosa, Mulet, Pitart, Rivera, Bou, Calvo, Cantón, González-López, Martínez-Martínez, Navarro, Oliver, Palacios-Baena, Pascual, Ruiz-Carrascoso, Vila, Aracil, Pérez-Vázquez, Oteo-Iglesias and the GEMARA/GEIRAS-SEIMC/REIPI CARB-ES-19 Study Group.)

Objectives: To quantify the workload and cost overload that the COVID-19 pandemic has meant for a Clinical Microbiology laboratory in a real-life scenario., Methods: We compared the number of samples received, their distribution, the human resources, and the budget of a Microbiology laboratory in the COVID pandemic (March-December 2020) with the same months of the previous year., Results: the total number of samples processed in the Clinical Microbiology laboratory in March to December 2020 increased 96.70% with respect to 2019 (from 246,060 to 483,993 samples), reflecting an increment of 127.50% when expressed as samples/1000 admissions (from 6057 to 13,780). The increase in workload was mainly at the expense of the virology (+2058%) and serology (+86%) areas. Despite additional personnel hiring, the samples processed per technician increased 12.5%. The extra cost attributed to Microbiology amounts to 6,616,511 euros (114.8%)., Conclusions: This is the first study to provide quantitative figures about workload and cost increase caused by the COVID-19 in a Microbiology laboratory., Competing Interests: Declaration of Competing Interests The authors report no conflicts of interest relevant to this article., (Copyright © 2021 Elsevier Inc. All rights reserved.)

In Spain, PCR is the tool of choice for the diagnosis of congenital Chagas disease (CD) and serology for diagnosing chronic CD. A loop-mediated isothermal amplification test for Trypanosoma cruzi DNA detection showed good analytical performance and ease of use. We aimed to evaluate the performance of the Loopamp Trypanosoma cruzi detection kit (Eiken Chemical Co. Ltd., Japan) ( Tcruzi -LAMP) for congenital and chronic CD diagnosis using well-characterized samples. We included samples from 39 congenital and 174 chronic CD cases and from 48 uninfected children born to infected mothers and 34 nonchagasic individuals. The sensitivity, specificity, and accuracy of Tcruzi -LAMP were estimated using standard case definitions for congenital CD (positive result by parasitological or PCR tests or serology after 9 months of age) and chronic CD (positive serology by at least two tests). The Tcruzi -LAMP results were read by visual examination and a real-time fluorimeter. For congenital CD, Tcruzi -LAMP sensitivity was 97% for both types of reading; specificity was 92% by visual examination and 94% by fluorimeter. For chronic CD, sensitivity was 47% and specificity 100%. The accuracy in congenital CD was >94% versus 56% in chronic CD. The agreement of Tcruzi -LAMP with PCR tests was better in congenital CD (kappa, 0.86 to 0.91) than in chronic CD (kappa, 0.67 to 0.83). The Loopamp Trypanosoma cruzi detection kit showed good performance for the diagnosis of congenital CD. Tcruzi -LAMP, like PCR, can be useful for the screening and early diagnosis of congenital infection., (Copyright © 2021 Flores-Chavez et al.)

Introduction: The chronic phase of Chagas disease (CD) is characterised by a low and intermittent parasitaemia. The Polymerase Chain Reaction (PCR) presents a variable sensitivity in this stage limiting its use as a diagnostic tool. Despite this, the use of PCR in untreated patients can provide information on the parasite behaviour and its presence in peripheral blood., Methods: A timely real-time PCR determination was performed on a cohort of 495 untreated chronic CD patients. Also, a subcohort of 29 patients was followed-up by serial real-time PCR during a period from 8 to 12 months in which they could not have access to the treatment due to lack of supply., Results: The positive percentage of real-time PCR in our series was 42%. Nevertheless, real-time PCR positive results were significantly higher in patients with five years or less of residence in Spain (P=.041). The detection of DNA was not related to the existence of cardiac and/or gastrointestinal abnormalities. In the follow-up subgroup, real-time PCR was consistently positive in 13.8% of patients, consistently negative in 31%, and intermittent in 55.2%., Conclusions: The different real-time PCR results regarding the time of residence suggests the possible relationship of external factors in the parasite presence in peripheral blood. On the other hand, specific host factors may be involved in the behaviour of parasitaemia over time., (Copyright © 2020 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.)

Aim: This work aimed to compare the sensitivity of four protocols for the detection of Trypanosoma cruzi DNA in 98 blood samples from chronic Chagas disease patients. Materials & methods: Two DNA extraction (automated and manual) methods and two T. cruzi satellite DNA qPCRs (with a recent design and the usually used set of primers) were analyzed. Results: Both DNA extraction methods and qPCR assays tested in this work gave comparable qualitative results, although the lowest Ct values were obtained when samples were analyzed using the new set of primers for T. cruzi satellite DNA. Conclusion: Our results encourage the implementation of automated DNA extraction systems and the new T. cruzi qPCR for the molecular diagnostics and treatment response monitoring of chronic Chagas disease patients.

The emergence of linezolid-resistant Enterococcus spp. (LRE) due to transferable resistance determinants is a matter of concern. To understand the contribution of the plasmid-encoded optrA and poxtA genes to the emergence of LRE, clinical isolates from different Spanish hospitals submitted to the Spanish Reference Laboratory from 2015-2018 were analysed. Linezolid resistance mechanisms were screened in all isolates by PCR and sequencing. Genetic relatedness of Enterococcus spp. carrying optrA and poxtA was studied by PFGE and MLST. Antimicrobial susceptibility was tested by broth microdilution using EUCAST standards. A total of 97 LRE isolates were studied, in 94 (96.9%) of which at least one resistance determinant was detected; 84/97 isolates (86.6%) presented a single resistance mechanism as follows: 45/84 (53.6%) carried the optrA gene, 38/84 (45.2%) carried the G2576T mutation and 1/84 (1.2%) carried the poxtA gene. In addition, 5/97 isolates (5.2%) carried both optrA and the G2576T mutation and 5/97 (5.2%) carried both optrA and poxtA. The optrA gene was more frequent in Enterococcus faecalis (83.6%) than Enterococcus faecium (11.1%) and was mainly associated with community-acquired urinary tract infections. Carriage of the poxtA gene was more frequent in E. faecium (13.9%) than E. faecalis (1.6%). Among the optrA-positive E. faecalis isolates, two main clusters were detected by PFGE. These two clusters belonged to ST585 and ST480 and were distributed throughout 11 and 6 Spanish provinces, respectively. This is the first description of LRE carrying the poxtA gene in Spain, including the co-existence of optrA and poxtA in five isolates., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: In recent years anti-TNF therapy has been associated with leishmaniasis in immunocompromised patients from endemic areas. Nevertheless, data on asymptomatic Leishmania infection in such patients is scarce. The aim of this study was to determine the prevalence of asymptomatic infection in inflammatory bowel disease (IBD) patients treated with TNF inhibitors living in an endemic area (Catalonia) and to follow up them to study how the infection evolved., Methods: 192 IBD patients (143 Crohn's disease; 49 ulcerative colitis) from Catalonia (Spain), an area endemic for L. infantum , were recruited. Peripheral blood samples were collected and tested for anti- Leishmania antibodies by Western blotting (WB). Leishmania kinetoplast DNA was detected in peripheral blood mononuclear cells (PBMC) by a quantitative PCR., Results: Serology was positive in 3.1% and Leishmania DNA was found in 8.8%, with a low parasitic load and humoral response. The prevalence was 10.9%, patients being considered infected if they tested positive by at least one of the techniques. Eight out of the 21 patients with asymptomatic leishmaniasis were monitored for 3-8 months after the first test. None of them showed an increased parasitemia or humoral response, or developed leishmaniasis during the follow-up period., Conclusion: The prevalence of Leishmania asymptomatic infection detected in our IBD cohort is similar to that found in healthy population in close endemic areas. Due to the short monitoring period, it is not possible to reach a conclusion about the risk of Leishmania reactivation from this study., (© 2020 The Author(s).)

Background: Antitrypanosomal treatment with Benznidazole (BZ) or Nifurtimox may be recommended for patients with chronic Chagas disease (CD) to reduce the onset or progression of symptoms. However, such treatment has limited efficacy and high level of toxic effects. In addition, the current cure biomarker (serology conversion) precludes any treatment assessment unless a prolonged follow-up is arranged. PCR is thus the most useful, alternative surrogate marker for evaluating responses to treatment. The aim of this study is to describe the usefulness of real-time PCR in monitoring BZ treatment within a large cohort of chronic CD cases in Barcelona., Methodology/principal Findings: A total of 370 chronic CD patients were monitored with real-time PCR post-BZ treatment. The median follow-up was 4 years (IQR 2.2-5.3y), with a median of 3 clinical visits (IQR 2-4). Only 8 patients (2.2%) presented with at least one incident of positive real-time PCR after treatment and were therefore considered as treatment failure. Four of those failure patients had completed full course treatment, whereas the remaining cases had defaulted with a statistical difference between both groups (p = 0.02). Half of the failure patients had undergone less than 4 years of follow-up monitoring all presented with parasitemia before treatment., Conclusions/significance: BZ treatment failure was highly infrequent in our cohort. BZ discontinuation was a risk factor for positive real-time PCR results during clinical follow-up. Regular testing with real-time PCR during follow-up allows for early detection of treatment failure in patients with chronic CD., Competing Interests: The authors have declared that no competing interests exist.

It is known that the immunoregulatory networks in human Chagas disease play a key role in parasitemia control during the acute phase. However, little is known regarding the control of parasitemia during the chronic phase. The aim of the study was to describe the serum cytokine profile of Trypanosoma cruzi chronically infected patients and to evaluate its relationship with the presence or absence of parasitemia in peripheral blood. This is a prospective observational study where adult Chagas disease patients were included. Patients previously treated for Chagas disease, pregnant women, and immunosuppressed patients were excluded. Demographic and clinical information was collected, and T. cruzi real-time polymerase chain reaction (RT-PCR) and serum cytokine profile were determined in peripheral blood. Forty-five patients were included. Trypanosoma cruzi RT-PCR in peripheral blood resulted positive in 19 (42.2%) patients. No differences in the serum cytokine profile were found depending on cardiac or digestive involvement. However, patients with positive T. cruzi RT-PCR had a higher median concentration of IL-10 and IL-1beta and a lower median concentration of IL-8 than those with negative T. cruzi PCR. These results reinforce the key role that this anti-inflammatory cytokine (IL-10) plays in parasitemia control.

Background: Chronic Chagas Disease (CD) diagnosis is based on serological methods employing crude, semipurified or recombinant antigens, which may result in low sensitivity or cross-reactivity. To reduce these restrictions, we developed a strategy involving use of molecules containing repetitive fragments of Trypanosoma cruzi conserved proteins. Diagnostic performance of IBMP-8.1 and IBMP-8.4 chimeric antigens (Molecular Biology Institute of Paraná - IBMP in Portuguese acronym) was assessed to diagnose T. cruzi-infected and non-infected immigrants living in Barcelona (Spain), a non-endemic setting for Chagas disease., Methods: Reactivity of IBMP-8.1 and IBMP-8.4 was assessed using an in-house automated ELISA with 347 positive and 331 negative individuals to Chagas disease. Antigenic cross-reactivity was measured with sera samples from pregnant women with Toxoplasma gondii (n = 98) and Zika virus (n = 75) antibodies., Results: The area under the curve values was 1 and 0.99 for the IBMP-8.1 and IBMP-8.4 proteins, respectively, demonstrating excellent diagnostic accuracy. The reactivity index was higher for IBMP-8.1 than IBMP-8.4 in positive samples and no significant difference in reactivity index was observed in negative samples. Sensitivity ranged from 99.4% for IBMP-8.1 to 99.1% for IBMP-8.4 and was not statistically different. Specificity for IBMP-8.1 reached 100 and 99.7% for IBMP-8.4, both nearly 100% accurate. No antigenic cross-reactivity was observed and reactivity index was similar to that for negative Chagas disease individuals., Conclusions: Our results showed an outstanding performance of IBMP-8.1 and IBMP-8.4 chimeric antigens by ELISA and suggest both chimeric antigens could also be used for Chagas disease diagnosis in immigrants living in non-endemic settings.

Introduction: Extrapulmonary tuberculosis (EPTB) is increasingly frequent in developed countries. When it is not clinically suspected, samples are not collected for culture and the only material available is a tissue paraffin block., Objective: The aim of this study was to evaluate FluoroType MTB (FT-MTB) and GenoType MTBDRplus methods for the detection of Mycobaterium tuberculosis complex in paraffin-embedded biopsies comparing the results to tuberculosis diagnosis., Methodology: A total of 17 paraffin-embedded tissues from different locations revealing granulomas were referred to the Mycobacteriology Laboratory and FT-MTB and GenoType MTBDRplus methods were performed. EPTB diagnosis was reached based on histologically compatible lesions, response to treatment and absence of alternative diagnosis. This case definition was considered gold standard for the assessment of the 2 molecular techniques performance., Results: Of the 17 individuals included in the study, 10 were clinically classified as EPTB and in 7 cases tuberculosis was ruled out. Of the 10 patients classified as EPTB, 6 (60%) obtained both FT-MTB and MTBDRplus positive results. Sensitivity and specificity were 60% and 71.4%, and 60% and 85.7% for FT-MTB and MTBDRplus, respectively., Conclusion: Molecular techniques might be useful tools for detection of Mycobaterium tuberculosis complex in paraffin-embedded biopsies especially when there is no sample available for culture.

Serodiscordance in Chagas disease (CD) remains a challenge since individuals with inconclusive results are clinically complicated to manage. This work, conducted outside the endemic area, aims to compare two different confirmatory techniques for the diagnosis of CD in individuals without a definitive diagnosis, to analyze the performance of the screening techniques in this group of patients, and to describe the serological follow-up of these subjects over time. Sera from 48 individuals with repeatedly discordant results by one recombinant enzyme immunoassay (r-ELISA) and one native ELISA (n-ELISA), were included in the study. Confirmatory procedures were performed through TESA-blot, using trypomastigote antigens of Trypanosoma cruzi, and in-house WB (IH-WB) using a lysate from T. cruzi epimastigotes. Of the 48 sera, TESA-blot confirmed 22 (45.8%) cases and IH-WB 17 (35.4%). Both techniques showed a substantial agreement (k = 0.604). Confirmation defined as the positivity of one of the ELISA and at least one of the confirmatory tests was reached in 24/48 (50%) cases. We found a great dispersion of r-ELISA index values, especially among individuals with confirmatory negative results, ranging from 0.03-6.2. Additionally, n-ELISA yielded a better performance than r-ELISA in this cohort of patients, showing a significantly greater agreement with the confirmatory methods. Our results indicate that either confirmatory test could be an efficient tool to solve inconclusive cases regardless of which form of the parasite's life cycle they use. Also, most individuals remain with discordant serology throughout the short-term follow-up period time of study. Finally, we consider that it is necessary to establish a reference test feasible and commercialized in all areas to solve the problem of inconclusive results., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Introduction: Campylobacter spp. infection is one of the leading causes of foodborne diarrhoeal illness in humans worldwide. The purpose of this study was to evaluate the DiaSorin LIAISON ® Campylobacter assay for human campylobacteriosis diagnosis., Methodology: A total of 645 stool samples from 640 patients suspected of having gastrointestinal infection were included. A stool culture was simultaneously performed with the DiaSorin LIAISON ® Campylobacter assay to detect the presence of Campylobacter spp., Results: Taking the conventional culture to be the perfect gold standard, sensitivity and specificity rates of the DiaSorin LIAISON ® Campylobacter assay were 100% and 97.7%, respectively; and 99.1% and 98.6%, respectively, when taking the culture to be the imperfect gold standard (Bayesian Model)., Conclusion: This new assay might be a useful tool especially for the screening of negative results., (Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.)

Reactivation of Chagas disease in the chronic phase may occur after solid organ transplantation, which may result in high parasitemia and severe clinical manifestations such as myocarditis and meningoencephalitis. The aim of the present study is to describe the prevalence of Chagas disease among solid organ-transplanted patients in a tertiary hospital from a nonendemic country. A cross-sectional study was performed at Vall d'Hebron University Hospital (Barcelona, Spain) from April to September 2016. Chagas disease screening was performed through serological tests in adult patients coming from endemic areas that had received solid organ transplantation and were being controlled in our hospital during the study period. Overall, 42 patients were included, 20 (47.6%) were male and median age was 50.5 (23-73) years. Transplanted organs were as follows: 18 kidneys, 17 lungs, and 7 livers. Three patients had Chagas disease, corresponding to a prevalence among this group of solid organ-transplanted patients of 7.1%. All three patients were born in Bolivia, had been diagnosed with Chagas disease and received specific treatment before the organ transplantation. We highly recommend providing screening tests for Chagas disease in patients with or candidates for solid organ transplantation coming from endemic areas, early treatment with benznidazole, and close follow-up to prevent clinical reactivations.

Objectives: In a previous study performed by our group, Strongyloides stercoralis infection in patients with Chagas disease was associated with higher proportion of Trypanosoma cruzi DNA detection in peripheral blood. The aim of the study was to confirm this association in a larger cohort of patients., Methods: Cross-sectional study of all patients with Chagas disease diagnosed from 2005 to 2015 during blood donation at the Catalan Blood Bank. Demographic data and T. cruzi RT-PCR were collected. S. stercoralis infection diagnosis was based on a serological test., Results: Two hundred and two blood donors were included. T. cruzi RT-PCR was positive in 72 (35.6%) patients, and S. stercoralis serology was positive in 22 (10.9%) patients. Patients with positive S. stercoralis serology had higher proportion of positive T. cruzi RT-PCR than those with negative serology (54.5% vs. 33.3%, P = 0.050), and the difference increased when taking a serological index cut-off of 2.5, which increases the specificity of the test to detect a confirmed strongyloidiasis (60% vs. 33%, P = 0.017)., Conclusions: Patients with Chagas disease with positive S. stercoralis serology had higher proportion of positive T. cruzi RT-PCR in peripheral blood than those with negative serology, which reflects the potential immunomodulatory effects of S. stercoralis in T. cruzi co-infected patients., (© 2017 John Wiley & Sons Ltd.)

Finding Dicrocoelium dendriticum eggs in human feces is exceptional and there are few prevalence data available. True infection occurs after accidental ingestion of ants containing metacercariae and spurious infection through the consumption of infected animal liver. Differential diagnosis between true and pseudo-infections is performed through stool examination after a diet free of liver. In addition, microscopy can help to differentiate the type of infection. We report six cases, all from sub-Saharan Africa, detection of this fluke at the Tropical Medicine Unit Vall d'Hebron-Drassanes (Barcelona, Spain). Dicrocoelium dendriticum transit eggs were visualized in five cases and there were no subsequent visualizations after diet, which reinforces that all these cases were false parasitism. In one case, few embryonated eggs were observed and the patient was treated for a possible true parasitism. There is a need to investigate the prevalence of D. dendriticum in our country focusing on the distinction between true and spurious infections., (© The American Society of Tropical Medicine and Hygiene.)

Parasitic diseases suppose an important health problem in people from high endemic areas, so these must be discarded properly. Usually, these infections develop asymptomatically but, in propitious situations, are likely to reactivate themselves and can cause clinical symptoms and/or complications in the receiving country. Moreover, in some cases it is possible local transmission. Early diagnosis of these parasitic diseases made by appropriate parasitological techniques and its specific treatment will benefit both, the individual and the community. These techniques must be selected according to geoepidemiological criteria, patient's origin, migration route or time spent outside the endemic area; but other factors must also be considered as its sensitivity and specificity, implementation experience and availability. Given the high prevalence of intestinal parasites on asymptomatic immigrants, it is recommended to conduct a study by coproparasitological techniques. Because of its potential severity, the screening of asymptomatic malaria with sensitive techniques such as PCR (polymerase chain reaction) is also advisable. Serological screening for Chagas disease should be performed on all Latin American immigrants, except for people from the Caribbean islands. Other important parasites, which should be excluded, are filariasis and urinary schistosomiasis, by using microscopic examination. The aim of this paper is to review the different techniques for the screening of parasitic diseases and its advices within the care protocols for asymptomatic immigrants., (Copyright © 2016 Elsevier España, S.L.U. All rights reserved.)