OBJECTIVES: We aimed to report the emergence of azole-resistant invasive aspergillosis in hematologic patients admitted to a tertiary hospital in Spain during the last 4 months. METHODS: Prospective, descriptive study was performed to describe and follow all consecutive proven and probable invasive aspergillosis resistant to azoles from hematological cohort during the last 4 months. All patients had fungal cultures and antifungal susceptibility or real-time PCR detection for Aspergillus species and real-time PCR detection for azole-resistant mutation. RESULTS: Four cases of invasive aspergillosis were diagnosed in 4 months. Three of them had azole-resistant aspergillosis. Microbiological diagnosis was achieved in three cases by means of fungal culture isolation and subsequent antifungal susceptibility whereas one case was diagnosed by PCR-based aspergillus and azole resistance detection. All the azole-resistant aspergillosis presented TR34/L98H mutation. Patients with azole-resistant aspergillosis had different hematologic diseases: multiple myeloma, lymphoblastic acute leukemia, and angioimmunoblastic T lymphoma. Regarding risk factors, one had prolonged neutropenia, two had corticosteroids, and two had viral co-infection. Two of the patients developed aspergillosis under treatment with azoles. CONCLUSION: We have observed a heightened risk of azole-resistant aspergillosis caused by A. fumigatus harboring the TR34/L98H mutation in patients with hematologic malignancies. The emergence of azole-resistant aspergillosis raises concerns for the community, highlighting the urgent need for increased surveillance and the importance of susceptibility testing and new drugs development.
Introduction: Malaria is one of the most prevalent infectious diseases in sub-Saharan Africa, with 247 million cases reported worldwide in 2021 according to the World Health Organization. Optical microscopy remains the gold standard technique for malaria diagnosis, however, it requires expertise, is time-consuming and difficult to reproduce. Therefore, new diagnostic techniques based on digital image analysis using artificial intelligence tools can improve diagnosis and help automate it. Methods: In this study, a dataset of 2571 labeled thick blood smear images were created. YOLOv5x, Faster R-CNN, SSD, and RetinaNet object detection neural networks were trained on the same dataset to evaluate their performance in Plasmodium parasite detection. Attention modules were applied and compared with YOLOv5x results. To automate the entire diagnostic process, a prototype of 3D-printed pieces was designed for the robotization of conventional optical microscopy, capable of auto-focusing the sample and tracking the entire slide. Results: Comparative analysis yielded a performance for YOLOv5x on a test set of 92.10% precision, 93.50% recall, 92.79% F-score, and 94.40% mAP0.5 for leukocyte, early and mature Plasmodium trophozoites overall detection. F-score values of each category were 99.0% for leukocytes, 88.6% for early trophozoites and 87.3% for mature trophozoites detection. Attention modules performance show non-significant statistical differences when compared to YOLOv5x original trained model. The predictive models were integrated into a smartphone-computer application for the purpose of image-based diagnostics in the laboratory. The system can perform a fully automated diagnosis by the auto-focus and X-Y movements of the robotized microscope, the CNN models trained for digital image analysis, and the smartphone device. The new prototype would determine whether a Giemsa-stained thick blood smear sample is positive/negative for Plasmodium infection and its parasite levels. The whole system was integrated into the iMAGING smartphone application. Conclusion: The coalescence of the fully-automated system via auto-focus and slide movements and the autonomous detection of Plasmodium parasites in digital images with a smartphone software and AI algorithms confers the prototype the optimal features to join the global effort against malaria, neglected tropical diseases and other infectious diseases.
The aim of this study was to validate the detection of anti-nucleocapsid protein (N protein) antibodies for the diagnosis of SARS-CoV-2 infection in light of the fact that most COVID-19 vaccines use the spike (S) protein as the antigen. Here, 3550 healthcare workers (HCWs) were enrolled from May 2020 (when no S protein vaccines were available). We defined SARS-CoV-2 infection if HCWs were found to be positive by RT-PCR or found to be positive in at least two different serological immunoassays. Serum samples from Biobanc I3PT-CERCA were analyzed by Roche Elecsys® (N protein) and Vircell IgG (N and S proteins) immunoassays. Discordant samples were reanalyzed with other commercial immunoassays. Roche Elecsys® showed the positivity of 539 (15.2%) HCWs, 664 (18.7%) were found to be positive by Vircell IgG immunoassays, and 164 samples (4.6%) showed discrepant results. According to our SARS-CoV-2 infection criteria, 563 HCWs had SARS-CoV-2 infection. The Roche Elecsys® immunoassay has a sensitivity, specificity, accuracy, and concordance with the presence of infection of 94.7%, 99.8%, 99.3%, and 0.96, respectively. Similar results were observed in a validation cohort of vaccinated HCWs. We conclude that the Roche Elecsys® SARS-CoV-2 N protein immunoassay demonstrated good performance in diagnosing previous SARS-CoV-2 infection in a large cohort of HCWs.
COVID-19 , Humans , COVID-19/diagnosis , SARS-CoV-2/genetics , COVID-19 Vaccines , Antibodies, Viral , Sensitivity and Specificity , Immunoassay/methods , Nucleocapsid Proteins , Immunoglobulin G , Vaccination
BACKGROUND: Pandemic preparedness is critical to respond effectively to existing and emerging/new viral pathogens. Important lessons have been learned during the last pandemic at various levels. This revision discusses some of the major challenges and potential ways to address them in the likely event of future pandemics. OBJECTIVES: To identify critical points of readiness that may help us accelerate the response to future pandemics from a clinical microbiology laboratory perspective with a focus on viral diagnostics and genomic sequencing. The potential areas of improvement identified are discussed from the sample collection to information reporting. SOURCES: Microbiologists and researchers from five countries reflect on challenges encountered during the COVID-19 pandemic, review published literature on prior and current pandemics, and suggest potential solutions in preparation for future outbreaks. CONTENT: Major challenges identified in the pre-analytic and post-analytic phases from sample collection to result reporting are discussed. From the perspective of clinical microbiology laboratories, the preparedness for a new pandemic should focus on zoonotic viruses. Laboratory readiness for scalability is critical and should include elements related to material procurement, training personnel, specific funding programmes, and regulatory issues to rapidly implement "in-house" tests. Laboratories across various countries should establish (or re-use) operational networks to communicate to respond effectively, ensuring the presence of agile circuits with full traceability of samples. IMPLICATIONS: Laboratory preparedness is paramount to respond effectively to emerging and re-emerging viral infections and to limit the clinical and societal impact of new potential pandemics. Agile and fully traceable methods for sample collection to report are the cornerstone of a successful response. Expert group communication and early involvement of information technology personnel are critical for preparedness. A specific budget for pandemic preparedness should be ring-fenced and added to the national health budgets.
Immune imprinting or original antigenic sin (OAS) is the process by which the humoral memory response to an antigen can inhibit the response to new epitopes of that antigen originating from a second encounter with the pathogen. Given the situation of the COVID-19 pandemic, multiple vaccines have been developed against SARS-CoV-2 infection. These vaccines are directed to the spike protein (S protein) of the original variant of Wuhan D614G. Vaccine memory immune response against S protein in noninfected subjects could inhibit, through the OAS mechanism, the response to new epitopes of SARS-CoV-2 after infection. The present study analyzes whether the memory antibody B cell response generated by mRNA vaccines against S protein can inhibit the primary antibody immune response to other SARS-CoV-2 antigens, such as nucleocapsid protein (N protein). SARS-CoV-2 primary infection in vaccinated healthcare workers (HCWs) produced significantly lower titers of anti-N antibodies than that in nonvaccinated HCWs: 5.7 (IQR 2.3-15.2) versus 12.2 (IQR 4.2-32.0), respectively (p = 0.005). Therefore, spike protein vaccine-induced immune imprinting (original antigenic sin) reduces N protein antibody response.
COVID-19 , Vaccines , Antibody Formation , COVID-19/prevention & control , Epitopes , Humans , Nucleocapsid Proteins , Pandemics , SARS-CoV-2 , Spike Glycoprotein, Coronavirus/genetics
The aim of this study was to characterize the antibody response induced by SARS-CoV-2 mRNA vaccines in a cohort of healthcare workers. A total of 2247 serum samples were analyzed using the Elecsys® Anti-SARS-CoV-2 S-test (Roche Diagnostics International Ltd., Rotkreuz, Switzerland). Sex, age, body mass index (BMI), arterial hypertension, smoking and time between infection and/or vaccination and serology were considered the confounding factors. Regarding the medians, subjects previously infected with SARS-CoV-2 who preserved their response to the nucleocapsid (N) protein showed higher humoral immunogenicity (BNT162b2: 6456.0 U/mL median; mRNA-1273: 2505.0 U/mL) compared with non-infected (BNT162b2: 867.0 U/mL; mRNA-1273: 2300.5 U/mL) and infected subjects with a lost response to N protein (BNT162b2: 2992.0 U/mL). After controlling for the confounders, a higher response was still observed for mRNA-1273 compared with BNT162b2 in uninfected individuals (FC = 2.35, p < 0.0001) but not in previously infected subjects (1.11 FC, p = 0.1862). The lowest levels of antibodies were detected in previously infected non-vaccinated individuals (39.4 U/mL). Clinical variables previously linked to poor prognoses regarding SARS-CoV-2 infection, such as age, BMI and arterial hypertension, were positively associated with increasing levels of anti-S protein antibody exclusively in infected subjects. The mRNA-1273 vaccine generated a higher antibody response to the S protein than BNT162b2 in non-infected subjects only.
COVID-19 , Hypertension , 2019-nCoV Vaccine mRNA-1273 , Antibodies, Viral , Antibody Formation , BNT162 Vaccine , COVID-19/prevention & control , Health Personnel , Humans , SARS-CoV-2/genetics , mRNA Vaccines
No disponible
Humans , Mycoplasma , Ureaplasma , Reproductive Tract Infections , Anus Diseases , Urethritis
BACKGROUND: STIs are a major public health concern. Screening programmes for asymptomatic users are key components of STI control. Traditional limitations of screening programmes include low population coverage and delays in treatments, thus reducing the expected impact on STI control. In our centre, the normal time from test to results was 4 days, and 7 days until treatment was established.To reduce time to treatment and to increase population coverage, we developed 'Drassanes Exprés', a testing service for asymptomatic STIs. The objectives of this study were to provide a guide for the implementation of a service with these characteristics and to evaluate the results of this intervention. METHODS: The Drassanes Exprés programme was launched in Spain on 07 November 2016 as a public, confidential and free-of-charge testing service for asymptomatic STIs, with same-day result notification. For this walk-in service, confidentiality was obtained by registering all information into the Laboratory Internal Software instead of the Electronic Patient Records. Samples were processed in a point-of-care laboratory and result notification was provided via mail or short message service.Information about workflow, screening protocols and result interpretation is detailed. Additionally, demographic characteristics, STI prevalence, and time from patients' sample collection to notification and treatment are analysed. RESULTS: Between 07 November 2016 and 07 November 2019, 13 993 users attended the Drassanes Exprés screening programme. Of these, 0.5% were transgender people, 29.3% women, 45.2% men who have sex with men and 25.1% men who have sex with women. The median age was 31 years (range: 26-39 years). Overall, 14.6% of users tested positive for at least one STI. The most prevalent infection was Chlamydia trachomatis (8.3%), followed by Neisseria gonorrhoeae (5.7%), syphilis (1.8%), HIV (0.4%) and hepatitis C virus (0.2%). The median time from test to results was 2.4 hours (range: 2-3.1 hours). Of 2049 users diagnosed with an STI, treatment was achieved in 97.0% of cases; the average time to treatment was 2.0 days. CONCLUSIONS: Drassanes Exprés is the first public programme for rapid, asymptomatic, STI screening and treatment in Spain. Assessing high-risk practices and providing confidentiality, easy access and rapid results/treatments are key elements in the development of STI screening programmes.
Chlamydia Infections , Gonorrhea , HIV Infections , Sexual and Gender Minorities , Sexually Transmitted Diseases , Adult , Chlamydia Infections/epidemiology , Chlamydia trachomatis , Female , Gonorrhea/diagnosis , Gonorrhea/drug therapy , Gonorrhea/epidemiology , HIV Infections/diagnosis , HIV Infections/epidemiology , HIV Infections/prevention & control , Homosexuality, Male , Humans , Male , Neisseria gonorrhoeae , Prevalence , Sexually Transmitted Diseases/diagnosis , Sexually Transmitted Diseases/drug therapy , Sexually Transmitted Diseases/epidemiology
Aim: To implement the multilocus sequence typing (MLST) methodology in syphilis samples previously characterized by enhanced CDC typing (ECDCT) and macrolide resistance. Materials & methods: MLST was performed on genital ulcer and blood samples by analyzing a region of the tp0136, tp0548 and tp0705loci using Sanger sequencing. Results: Up to 59/85 (69.4%) of genital ulcer and 4/39 (10.3%) of whole blood samples were fully typed. The most frequent profiles were 1.3.1 (56%) and 1.1.1 (11%). All the 1.3.1 samples typed carried the A2058G mutation, responsible for macrolide resistance. MLST and ECDCT showed similar overall typing yields. Conclusion: Several allelic profiles of T. pallidum subsp. pallidum were identified and classified into two major genetic clades in Barcelona. Our results were similar to that described in Europe.
Anti-Bacterial Agents , Drug Resistance, Bacterial , Syphilis/microbiology , Treponema/classification , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial/genetics , Humans , Macrolides/pharmacology , Multilocus Sequence Typing , Spain , Ulcer
Mycoplasma Infections , Mycoplasma genitalium , Sexually Transmitted Diseases , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Drug Resistance, Bacterial , France , Humans , Macrolides/pharmacology , Male , Mycoplasma Infections/drug therapy , Mycoplasma genitalium/genetics , Prevalence , Sexually Transmitted Diseases/drug therapy
Mycoplasma pneumoniae is a bacterial human pathogen that causes primary atypical pneumonia. M. pneumoniae motility and infectivity are mediated by the immunodominant proteins P1 and P40/P90, which form a transmembrane adhesion complex. Here we report the structure of P1, determined by X-ray crystallography and cryo-electron microscopy, and the X-ray structure of P40/P90. Contrary to what had been suggested, the binding site for sialic acid was found in P40/P90 and not in P1. Genetic and clinical variability concentrates on the N-terminal domain surfaces of P1 and P40/P90. Polyclonal antibodies generated against the mostly conserved C-terminal domain of P1 inhibited adhesion of M. pneumoniae, and serology assays with sera from infected patients were positive when tested against this C-terminal domain. P40/P90 also showed strong reactivity against human infected sera. The architectural elements determined for P1 and P40/P90 open new possibilities in vaccine development against M. pneumoniae infections.
Adhesins, Bacterial/immunology , Bacterial Adhesion/immunology , Mycoplasma pneumoniae/immunology , Pneumonia, Mycoplasma/immunology , Adhesins, Bacterial/isolation & purification , Adhesins, Bacterial/ultrastructure , Cryoelectron Microscopy , Crystallography, X-Ray , Mycoplasma pneumoniae/isolation & purification , Mycoplasma pneumoniae/pathogenicity , Pneumonia, Mycoplasma/blood , Pneumonia, Mycoplasma/microbiology , Protein Domains/immunology
Antibiotic resistance in Mycoplasma genitalium (MG) is rising globally, especially to macrolides. In response to this challenge, assays reporting both the detection of MG and macrolide resistance-mediating mutations (MRMM) allow therapy to be tailored to the individual. The study evaluated the performance of the ResistancePlus® MG FleXible assay for the detection of MG and MRMM. Overall, the test performed well for the detection of MG compared to the AllplexTM STI Essential assay, used as a reference, with a kappa value of 0.926 (95% CI, 0.863-0.990). The kit also performed well for the detection of MRMM when compared with Sanger sequencing of the 23S rRNA gene, with a kappa value of 0.901 (95% CI, 0.807-0.996). The rate of MRMM in MG among the study population was 41.8%. In conclusion, the ResistancePlus® MG FleXible is a rapid, simple, and accurate cartridge-based assay for simultaneous detection of MG and MRMM in clinical settings.
Bacteriological Techniques , Drug Resistance, Bacterial/genetics , Macrolides/pharmacology , Molecular Diagnostic Techniques , Mycoplasma Infections/diagnosis , Mycoplasma genitalium/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacteriological Techniques/standards , Humans , Molecular Diagnostic Techniques/standards , Mutation , Mycoplasma Infections/epidemiology , Mycoplasma genitalium/genetics , Prevalence , RNA, Ribosomal, 23S/genetics , Reagent Kits, Diagnostic , Sensitivity and Specificity , Spain/epidemiology
INTRODUCTION: Mycoplasma genitalium is a major cause of urethritis and other genital syndromes. Antibiotic resistance, especially to macrolides, is increasing at an alarming rate worldwide. The aim of this study was to estimate the rate of macrolide resistance in M. genitalium among a 2016-2017 cohort of patients in Barcelona, Spain; and to compare this estimate with previous data from 2013 to 2014 in this region. METHODS: The study was conducted retrospectively with M. genitalium-positive samples collected between December 2016 and February 2017 at the Hospital Vall d'Hebron Microbiology Department. Genotypic markers of macrolide resistance were primarily detected using the ResistancePlus(R) MG molecular assay (SpeeDx). Mutations were then confirmed by sequencing. RESULTS: Macrolide resistance-mediating mutations were detected in 30/83 infections (36.1% [95% CI, 25.9%-47.4%]). This resistance was more frequent among men who have sex with men (55.0% [95% CI, 38.5%-70.7%]) compared to heterosexual men (27.3% [95% CI, 10.7%-50.2%]) and women (9.5% [95% CI, 1.3%-30.4%]), p < 0.001. Additionally, macrolide resistance did not significantly increase in this cohort when compared with previous investigations. CONCLUSION: Despite the current notable rate of macrolide resistance in M. genitalium, resistance did not significantly increase between 2013-2014 and 2016-2017 in our region. Nevertheless, strict local surveillance and the implementation of rapid diagnostic tests that combine the detection of the bacterium and resistance-mediating mutations may facilitate the optimization of antibiotic administration and reduce the transmission of resistance in M. genitalium
INTRODUCCIÓN: Mycoplasma genitalium es causa de uretritis y otras enfermedades genitales. Las resistencias antibióticas, especialmente a macrólidos, están aumentando de forma alarmante a nivel mundial. El objetivo del estudio fue estimar la tasa de resistencia a macrólidos en M. genitalium sobre una cohorte de pacientes entre los años 2016-2017 en Barcelona, España; y comparar esta estimación con datos previos de 2013-2014 en esta región. MÉTODOS: El estudio se realizó de forma retrospectiva sobre muestras positivas para M. genitalium recogidas entre diciembre 2016 y febrero 2017 en el Departamento de Microbiología del Hospital Vall d'Hebron. Los marcadores genotípicos de resistencia a macrólidos se detectaron en primer lugar con el ensayo molecular ResistancePlus(R) MG (SpeeDx). Las mutaciones se confirmaron posteriormente por secuenciación. RESULTADOS: Se detectaron mutaciones asociadas a resistencia a macrólidos en 30/83 (36,1% [IC 95%: 25,9-47,4%]) infecciones. Esta resistencia fue más frecuente en hombres que tienen sexo con hombres (55,0% [IC 95%: 38,5-70,7%]) comparada con la tasa en hombres heterosexuales (27,3% [IC 95%: 10,7-50,2%]) y mujeres (9,5% [IC 95%: 1,3-30,4%]), p < 0,001. Además, la resistencia a macrólidos no aumentó significativamente en esta serie en comparación con investigaciones previas. CONCLUSIÓN: A pesar de la tasa notable de resistencia a macrólidos en M. genitalium, esta no aumentó significativamente entre los años 2013-14 y 2016-17 en nuestro entorno. No obstante, una estricta vigilancia a nivel local junto con la implementación de pruebas diagnósticas rápidas que combinan la detección de la bacteria y las mutaciones de resistencia puede facilitar la optimización de la administración antibiótica y reducir la transmisión de resistencias en M. genitalium
Humans , Female , Adolescent , Young Adult , Adult , Middle Aged , Drug Resistance, Bacterial/genetics , Mutation/genetics , Macrolides/pharmacology , Mycoplasma genitalium/drug effects , Mycoplasma genitalium/genetics , Mycoplasma Infections/microbiology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Retrospective Studies , Cohort Studies , Spain
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Fluoroquinolones/pharmacology , Macrolides/pharmacology , Mycoplasma Infections/drug therapy , Mycoplasma Infections/microbiology , Mycoplasma genitalium/drug effects , Anti-Bacterial Agents/therapeutic use , DNA, Bacterial/analysis , Fluoroquinolones/therapeutic use , Humans , Macrolides/therapeutic use , Mycoplasma genitalium/genetics , Mycoplasma genitalium/isolation & purification
Sexually transmitted infections (STIs) by Mycoplasma genitalium are a major problem worldwide, especially given their marked and rapid propensity for developing antimicrobial resistance. Since very few treatment options exist, clinicians face an important challenge in the management of the infection. In this scenario, little is known regarding the transmission dynamics of M. genitalium and the epidemiology of antimicrobial resistance. This mgpB-based molecular typing study, conducted among 54 asymptomatically infected individuals prospectively recruited from an STI screening service, reveals two distinct epidemiological clusters that significantly correlate with sexual conduct in heterosexuals and men who have sex with men (MSM), respectively. This well-defined structuration suggests the presence of two independent sexual networks with little connectivity between them. On the other hand, the study demonstrates the multiclonal feature of the emergence of antibiotic resistance in M. genitalium to both macrolides and fluoroquinolones. The high prevalence of macrolide resistance in M. genitalium among MSM, influenced by dense network connectivity and strong antibiotic selective pressure, may correspond to allodemics affecting other STIs such as gonorrhea, syphilis and enteric pathogens. Collaterally, the structural and functional impact of mutations in the mgpB gene, encoding the major adhesin P140 (MgpB), may require further investigation.
Mycoplasma Infections , Mycoplasma genitalium , Sexual and Gender Minorities , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Drug Resistance, Bacterial , Homosexuality, Male , Humans , Macrolides/pharmacology , Male , Mycoplasma Infections/epidemiology , Mycoplasma genitalium/genetics , Prevalence
INTRODUCTION: Mycoplasma genitalium is a major cause of urethritis and other genital syndromes. Antibiotic resistance, especially to macrolides, is increasing at an alarming rate worldwide. The aim of this study was to estimate the rate of macrolide resistance in M. genitalium among a 2016-2017 cohort of patients in Barcelona, Spain; and to compare this estimate with previous data from 2013 to 2014 in this region. METHODS: The study was conducted retrospectively with M. genitalium-positive samples collected between December 2016 and February 2017 at the Hospital Vall d'Hebron Microbiology Department. Genotypic markers of macrolide resistance were primarily detected using the ResistancePlus® MG molecular assay (SpeeDx). Mutations were then confirmed by sequencing. RESULTS: Macrolide resistance-mediating mutations were detected in 30/83 infections (36.1% [95% CI, 25.9%-47.4%]). This resistance was more frequent among men who have sex with men (55.0% [95% CI, 38.5%-70.7%]) compared to heterosexual men (27.3% [95% CI, 10.7%-50.2%]) and women (9.5% [95% CI, 1.3%-30.4%]), p<0.001. Additionally, macrolide resistance did not significantly increase in this cohort when compared with previous investigations. CONCLUSION: Despite the current notable rate of macrolide resistance in M. genitalium, resistance did not significantly increase between 2013-2014 and 2016-2017 in our region. Nevertheless, strict local surveillance and the implementation of rapid diagnostic tests that combine the detection of the bacterium and resistance-mediating mutations may facilitate the optimization of antibiotic administration and reduce the transmission of resistance in M. genitalium.
Anti-Bacterial Agents , Drug Resistance, Bacterial , Macrolides/pharmacology , Mycoplasma Infections , Mycoplasma genitalium , Sexual and Gender Minorities , Anti-Bacterial Agents/pharmacology , Diagnostic Tests, Routine , Drug Resistance, Bacterial/genetics , Female , Homosexuality, Male , Humans , Male , Mutation , Mycoplasma Infections/microbiology , Mycoplasma genitalium/drug effects , Mycoplasma genitalium/genetics , Retrospective Studies , Spain
