Rapid syndromic molecular testing and human parechovirus infection in children: A report of three cases in Argentina.

Human parechovirus (HPeV) is one of the members of the family Picornaviridae that has been associated with fever of unknown origin, gastroenteritis, clinical sepsis, meningitis, or encephalitis in very young infants. HPeV detection is not routinely performed in most clinical microbiology laboratories in Argentina and, therefore, its real prevalence is unknown. We here report three cases of HPeV CNS infection that presented to our hospital with different clinical features after the implementation of a multiplex PCR meningitis/encephalitis panel. Molecular diagnostic techniques could help improve patient care and understand the real prevalence of this infection in Argentina.

[Detección y caracterización molecular de virus respiratorios causantes de infección respiratoria aguda en población adulta].

INTRODUCTION: Acute respiratory infections are one of the main causes of morbidity and mortality in older adults and patients with chronic diseases. Among the responsible etiological agents are human respiratory viruses, such as: respiratory syncytial virus, parainfluenza virus and metapneumovirus. OBJECTIVE: To carry out a differential diagnostic study of respiratory viruses circulating and co-circulating in an adult population. METHODS: A pilot study was conducted in patients older than 18 years, who presented signs and symptoms suggestive of acute respiratory infection and whose clinical picture did not exceed 15 days of evolution; end-point polymerase chain reaction assays were performed with the use of specific oligonucleotides for molecular diagnosis. RESULTS: 72 specimens of patients with an age of 51.33 ± 19.33 years, with a predominance of females (4.5:1); original inhabitants of Mexico City; only 22 were positive for respiratory viruses, being mostly metapneumovirus infections. CONCLUSIONS: The knowledge of the circulating viral strains in the population will allow to determine changes that can declare an epidemiological alert leading to the best decision making for the benefit of the patients.

INTRODUCCIÓN: Las infecciones respiratorias agudas son una de las principales causas de morbimortalidad en adultos mayores y pacientes con enfermedades crónicas. Dentro de los agentes etiológicos responsables se encuentran los virus respiratorios humanos, tales como: virus sincitial respiratorio, virus parainfluenza y metapneumovirus. OBJETIVO: Realizar un estudio de diagnóstico diferencial de virus respiratorios que circulan y cocirculan en una población adulta. MÉTODO: Se realizó un estudio tipo piloto en pacientes mayores de 18 años, que presentaron signos y síntomas sugestivos de infección respiratoria aguda y cuyo cuadro clínico no sobrepasara los 15 días de evolución; se realizaron ensayos de reacción en cadena de la polimerasa de punto final con el uso de oligonucleótidos específicos para el diagnóstico molecular. RESULTADOS: Se tipificaron 72 especímenes de pacientes con una edad de 51.33 ± 19.33 años, con predominio del sexo femenino (4.5:1); originarios en su totalidad de la Ciudad de México; solo 22 fueron positivos para virus respiratorios, siendo en su mayoría infecciones por metapneumovirus. CONCLUSIONES: El conocimiento de las cepas virales circulantes en la población permitirá determinar cambios que puedan declarar una alerta epidemiológica llevando a la mejor toma de decisiones en beneficio de los pacientes.

Prevalence of Trichomonas vaginalis and Human papillomavirus in female sex workers in Central Veracruz, Mexico.

Female sex workers (FSWs) have been considered a key population for sexually transmitted infections (STIs); therefore, they are periodically screened as a requirement to obtain a work card. However, there is insufficient epidemiological data on STIs among FSWs in Mexico. The detection of Trichomonas vaginalis is limited to microscopic studies and the molecular screening of Human papillomavirus (HPV) is only done to women 35 years of age and older. The objective of this study was to determine the prevalence of T. vaginalis and HPV infections in FSWs in the city of Orizaba, Veracruz, Mexico. Samples from 105 FSWs were obtained by cervical swab and analyzed. The identification of T. vaginalis and HPV was performed by molecular methods. HPV DNA was identified in 5.71% of the samples with the presence of HPV16, HPV18, and HPV58. A percentage of 25.7% samples were positive for T. vaginalis for optical microscopy and 23.8% for PCR. The results of the study indicate the need to incorporate more sensitive methods for the timely diagnosis of STIs as well as comprehensive health promotion programs directed to the most vulnerable groups among FSWs.

Molecular methods for septicemia diagnosis. / Métodos moleculares para el diagnóstico de septicemia.

Septicemia remains a major cause of hospital mortality. Blood culture remains the best approach to identify the etiological microorganisms when a bloodstream infection is suspected but it takes long time because it relies on bacterial or fungal growth. The introduction in clinical microbiology laboratories of the matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology, DNA hybridization, microarrays or rapid PCR-based test significantly reduce the time to results. Tests for direct detection in whole blood samples are highly desirable because of their potential to identify bloodstream pathogens without waiting for blood cultures to become positive. Nonetheless, limitations of current molecular diagnostic methods are substantial. This article reviews these new molecular approaches (LightCycler SeptiFast, Magicplex sepsis real time, Septitest, VYOO, PCR/ESI-MS analysis, T2Candida).

Evaluation of the Luminex xTAG®-GPP (Gastrointestinal Pathogen Panel) in the diagnosis of diseases with acute diarrhoea. / Evaluación del panel gastrointestinal xTAG®-GPP de Luminex en el diagnóstico de las gastroenteritis agudas.

INTRODUCTION: Most Microbiology laboratories use different techniques for the diagnosis of gastrointestinal infections. Some of which require at least 72 hours to obtain final results. MATERIAL AND METHODS: The gastrointestinal panel Luminex (xTAG-GPP, Luminex Molecular Diagnostics, Toronto, Canada) is a qualitative multiplex fast and sensitive assay able to detect and to identify the 15 most common pathogens causing gastrointestinal infection simultaneously. We evaluated this multiplex panel comparing it with conventional methods used in our laboratory. RESULTS: We analyzed 225 samples of feces. Through the conventional methods were positive 74 samples (32.9%). Through the Luminex method were positive 137 samples (60.9%). CONCLUSIONS: The use of the xTAG® GPP system in Clinical Microbiology can improve the diagnosis of gastrointestinal infectious because it provides results in less than 8 hours. Some pathogens should be applied with caution and should be interpreted based on the patient́s clinical data.

Evaluation of a commercial multiplex PCR (Unyvero i60®) designed for the diagnosis of bone and joint infections using prosthetic-joint sonication.

BACKGROUND: The development of sonication protocols over the last few years has improved the sensitivity of conventional cultures for the diagnosis of prosthetic-joint infection (PJI). However, the development of a new, specifically designed kit for the molecular diagnosis of PJI could provide a major improvement in this field. METHODS: Prostheses retrieved from patients who underwent implant removal from May 2014 to May 2015 were sent for culture, and processed according to a previously defined protocol that included sonication. Furthermore, 180 microlitres of sonication fluid were used to carry out the multiplex PCR test (Unyvero i60 system®). A comparison of the sensitivity, specificity, positive (PPV) and negative (NPV) predictive value, was performed. The study was approved by the Clinical Research Ethics Committee. RESULTS: The analysis included 88 prostheses from 68 patients (1.29 prostheses/patient). The type of prostheses studied were knee (n=55), total hip (n=26), partial hip (n=5), and shoulder (n=2). Twenty-nine patients were diagnosed with a PJI (15 delayed, 12 acute, and 2 haematogenous infections). In 24 cases, the result of the PCR was positive, all but 1 corresponding to patients with clinical criteria of PJI. Nine resistance mechanisms were detected from 5 samples. The Unyvero i60 system® showed slightly better results than traditional culture in terms of specificity and PPV. CONCLUSIONS: The Unyvero i60 system® may play a role in rapid diagnosis of PJI, due to its high specificity and PPV. However, despite these results, cultures have to be performed to detect organisms not detected by the system.

Molecular detection of Sarcocystis aucheniae in the blood of llamas from Argentina.

Sarcocystis aucheniae are apicomplexan protozoa that infect South American camelids (SACs), giving rise to macroscopic cysts similar to rice grains in skeletal muscles. Visual detection of macrocysts in slaughtered animals hampers commercialization of SAC meat, a highly relevant economic exploitation for Andean rural families. Importantly, the consumption of undercooked S. aucheniae-infested meat causes gastroenteritis. A carnivore definitive host, possibly the dog, acquires the parasite when feeding on infected SAC meat, and later eliminates infective oocysts in its feces. The parasite cycle is completed when SACs ingest contaminated water or pastures. We hypothesized that parasite DNA can be detected in SAC blood using molecular methods. In order to test this hypothesis, a seminested PCR format was specifically designed to target the hypervariable 18S rRNA gene region of S. aucheniae. PCR conditions were optimized using genomic DNA extracted from macrocyst bradyzoites. A detection limit of up to 1 parasite in 10µl of llama blood was established based on DNA samples extracted from aliquots of S. aucheniae bradyzoite-spiked non-infected llama blood. The seminested PCR allowed to detect natural infections of S. aucheniae in llama blood samples originating in the Andean flatlands of Argentina. Specific amplification of S. aucheniae DNA was corroborated by amplicon sequencing. This is the first report of S. aucheniae detection in llama blood, which provides a valuable diagnostic tool for epidemiological studies and for the evaluation of the efficacy of control measures for this parasitosis.

[RECIST and the radiologist]. / RECIST y el radiólogo.

Biomedical imaging makes it possible not only to diagnose and stage cancer, but also to follow up patients and evaluate the response to treatment. RECIST (Response Evaluation Criteria In Solid Tumors) provides a method to monitor the response to treatment based on one dimensional measurements of tumors obtained with reproducible imaging techniques like CT, MRI, and PET. The metabolic changes induced by new treatments modify the biology and behavior of the tumor; occasionally, there is a discrepancy between the patient's clinical condition and the response measured by RECIST, which indicates that functional tests need to be included in the evaluation of the response to treatment. The objective is to review the RECIST criteria to include the contribution of functional imaging to enable the efficacy and effects of the treatment in patients with solid tumors.

Expert consensus of the Spanish Society of Pathology and the Spanish Society of Medical Oncology on the determination of biomarkers in pancreatic and biliary tract cancer.

Pancreatic cancer and biliary tract cancer have a poor prognosis. In recent years, the development of new diagnostic techniques has enabled the identification of the main genetic alterations involved in the development of these tumours. Multiple studies have assessed the ability to predict response to treatment of certain biomarkers, such as BRCA in pancreatic cancer, IDH1 or FGFR2 in biliary tract cancer and microsatellite instability or NTRK fusions in an agnostic tumour fashion. In this consensus, a group of experts selected by the Spanish Society of Medical Oncology (SEOM) and the Spanish Society of Pathology (SEAP) reviewed the role played by these mutations in the process of carcinogenesis and their clinical implications. Based on their results, a series of recommendations are made to optimize the determination of these biomarkers and thus help standardize the diagnosis and treatment of these tumours.

Clinical evaluation of a new molecular method for the detection of multidrug-resistant microorganisms.

INTRODUCTION: The main objective of this work is to carry out the clinical validation of the trial with the AMR Direct Flow Chip starting from either nasal swabs, rectal swabs directly or from isolated strains to detect antibiotic resistance genes. METHODS: We developed the preclinical validation of the assay with 104 known bacterial isolates. A total of 210 nasal or rectal swab samples were analyzed. The AMR assay is based on multiplex PCR followed by reverse dot blot hybridization on DNA arrays fully automated by using the HS24 platform. RESULTS: Both the sensitivity and specificity of the preclinical assay were 100%, with the 104 samples correctly identified. In the clinical validation, the sensitivity was 100% and the specificity was between 100% in nasal swabs and 97% in rectal swabs. CONCLUSIONS: The AMR Direct Flow Chip® is a rapid and effective assay for the detection of multidrug-resistant microorganisms (MDR) from nasal and rectal swab samples.

Molecular diagnosis of the central nervous system (CNS) infections.

Central nervous system (CNS) infections such as meningitis and encephalitis are medical emergencies that require rapid diagnosis of the causative pathogen to guide early and adequate treatment since a delay in implementing an adequate antimicrobial therapy can lead to death. The current microbiological diagnostic methods based on culture or antigen detection have important limitations in their capacity to accurately identify the different potential pathogens causing CNS and, in the time, to obtaining results. Rapid syndromic molecular arrays have been developed. The main advantage of using a meningoencephalitis panel based in a multiplex test is that includes bacteria, viruses and fungi, covering the most prevalent microorganisms causing meningitis and encephalitis and the turn-around time is circa 1h. The use of these multiplex-PCR based tools is reviewed and the advantages and disadvantages of this technique are discussed.

Clinical evaluation of a new molecular method for the detection of multidrug-resistant microorganisms. / Evaluación clínica de un nuevo método molecular para la detección de microorganismos multirresistentes.

INTRODUCTION: The main objective of this work is to carry out the clinical validation of the trial with the AMR Direct Flow Chip® starting from either nasal swabs, rectal swabs directly or from isolated strains to detect antibiotic resistance genes. METHODS: We developed the preclinical validation of the assay with 104 known bacterial isolates. A total of 210 nasal or rectal swab samples were analyzed. The AMR assay is based on multiplex PCR followed by reverse dot blot hybridization on DNA arrays fully automated by using the HS24 platform. The completion time of the full analysis is 3 hours. RESULTS: Both the sensitivity and specificity of the preclinical assay were 100%, with the 104 samples correctly identified. In the clinical validation, the sensitivity was 100% and the specificity was between 100% in rectal swabs and 97% in nasal swabs. CONCLUSIONS: The AMR Direct Flow Chip® is a rapid and effective assay for the detection of multidrug-resistant microorganisms from nasal and rectal swab samples.

Molecular diagnosis of the central nervous system (CNS) infections.

Central nervous system (CNS) infections such as meningitis and encephalitis are medical emergencies that require rapid diagnosis of the causative pathogen to guide early and adequate treatment since a delay in implementing an adequate antimicrobial therapy can lead to death. The current microbiological diagnostic methods based on culture or antigen detection have important limitations in their capacity to accurately identify the different potential pathogens causing CNS and, in the time, to obtaining results. Rapid syndromic molecular arrays have been developed. The main advantage of using a meningoencephalitis panel based in a multiplex test is that includes bacteria, viruses and fungi, covering the most prevalent microorganisms causing meningitis and encephalitis and the turn-around time is circa 1h. The use of these multiplex-PCR based tools is reviewed and the advantages and disadvantages of this technique are discussed.

Rhodotorula dairenensis fungemia in a patient with cancer.

BACKGROUND: Rhodotorula species were traditionally considered non-virulent environmental microorganisms, but are nowadays considered important human pathogens, especially in immunocompromised individuals. CASE REPORT: We present the case of a 73 year-old man with diarrhea, anorexia and fever. In the blood analyses, both aerobic blood culture bottles yielded the growth of Rhodotorula dairenensis. The MALDI-TOF MS score was inadequate to provide an identification, which was achieved by means of molecular techniques. Treatment with an echinocandin was started, but the patient died. CONCLUSIONS: Basidiomycetous yeast genera such as Rhodotorula can cause invasive and severe infections, e.g., fungemia, especially in patients with central venous catheter or another indwelling device.

Diagnóstico molecular de parasitosis intestinales.

Infections causes by parasites of the gastrointestinal tract are a global public health problem. In industrialised countries, their particular epidemiological (low general prevalence of enteroparasites), economic (high labour costs) and clinical characteristics (constant increase in the number of samples and diagnostic determinations to be performed) have led molecular techniques to progressively replace conventional microscopy as the first-line diagnostic method of these pathogens in modern clinical laboratories. PCR-based techniques, particularly those developed for the simultaneous detection of the various agents that can cause the same infectious disease (syndromic diagnosis), already represent a cost-effective option that allow process automisation, workflow optimisation, and comparison of results among different laboratories, and facilitate accreditation of diagnostic procedures. This review clearly and concisely discusses the current situation of the molecular diagnosis of the main species of intestinal parasites in humans, particularly the enteric protozoans causing diarrhoea (Cryptosporidium spp., Giardia duodenalis, Entamoeba histolytica), the most important members the Microsporidia phyla (Enterocytozoon bieneusi) and Stramenopiles phyla (Blastocystis sp.), as well as the helminths transmitted by soil (Ancylostoma spp., Ascaris lumbricoides, Necator americanus, Strongyloides stercoralis and Trichuris trichiura) and food (Anisakis spp., Clonorchis sinensis, Fasciola spp., Taenia solium, and Trichinella spiralis). Special attention is paid to the description of available techniques and formats, to their diagnostic benefits and the most widely used genetic markers for their detection, both in clinical laboratories and genotyping in referral and research centres.

Actualización en la detección de la tuberculosis a través de pruebas moleculares / An update on the detection of tuberculosis through molecular testing

La tuberculosis es una enfermedad infecciosa respiratoria que afecta a un tercio de la población mundial y es una amenaza significativa para la salud global. La detección de la tuberculosis de manera temprana es crucial para un tratamiento eficaz y prevenir su propagación. Una solución para mejorar el diagnóstico y abordar la resistencia a los medicamentos antituberculosos es el uso de pruebas moleculares de alto rendimiento para la identificación del Mycobacterium tuberculosis y su susceptibilidad. Este estudio de revisión narrativa busca describir las generalidades, la eficacia, la sensibilidad, las ventajas y las limitaciones de las principales pruebas moleculares; Truenat® MTB, MTB plus y MTB-RIF, Abbott RealTime MTB y MTB RIF/INH en el sistema m2000sp y m2000rt y FluoroType MTBDR, además, de compararlas con GeneXpert MTB/RIF o Xpert Ultra, utilizadas para la detección del patógeno resistente a medicamentos tuberculosos. Estas pruebas utilizan diversas técnicas para la detección del ADN del Mycobacterium tuberculosis y la cuantificación de la carga bacteriana con alta sensibilidad y especificidad, resultados rápidos, reducción de los errores humanos, así como la detección temprana de cepas drogo-resistentes. A pesar de que requieren infraestructura especializada y competencias profesionales para su implementación, representan avances significativos con el potencial de mejorar la atención sanitaria y la gestión de la tuberculosis. Estas pruebas moleculares, comparadas con el GeneXpert, son una alternativa viable, aunque esta última tecnología sigue siendo la preferida en áreas con recursos limitados

Tuberculosis is a respiratory infectious disease that affects one third of the world's population and is a significant threat to global health. Detecting tuberculosis early is crucial for effective treatment and preventing its spread. One solution to improve diagnosis and address antituberculosis drug resistance is the use of high-throughput molecular tests for the identification of Mycobacterium tuberculosis and its susceptibility. This narrative review study seeks to describe the generalities, efficacy, sensitivity, advantages and limitations of the main molecular tests: Truenat® MTB, MTB plus and MTB-RIF, Abbott RealTime MTB and MTB RIF/INH on the m2000sp and m2000rt system and FluoroType MTBDR, and to compare them with GeneXpert MTB/RIF or Xpert Ultra, used for the detection of the tuberculosis drug-resistant pathogen. These tests use various techniques for the detection of Mycobacterium tuberculosis DNA and quantification of bacterial load with high sensitivity and specificity, rapid results, reduction of human error, as well as early detection of drug-resistant strains

Use of the rapid molecular test for tuberculosis among people deprived of liberty: a scoping review / Uso de la prueba molecular rápida de tuberculosis en personas privadas de libertad: revisión de alcance / Utilização do teste rápido molecular para tuberculose entre pessoas privadas de liberdade: revisão de escopo

ABSTRACT Objective: to map the repercussions of using the rapid molecular test for diagnosing tuberculosis among people deprived of liberty in the scientific literature. Method: this is a scoping review following the recommendations of the Joanna Briggs Institute and PRISMA for Scoping Reviews. The search was conducted using controlled and free vocabulary in the following databases: EMBASE, Scopus, MEDLINE, Cinahl, Academic Search Premier, LILACS and Web of Science, in the Brazilian Digital Library of Theses and Dissertations and Google Scholar. The materials which answered the review question were selected by two independent reviewers based on reading the titles, abstracts and publications. All types of studies and publications were included. The extracted data was subjected to narrative synthesis and presented graphically. Results: a total of 13 among the 461 publications found were included in the review. The studies pointed out the following repercussions of using the rapid molecular test in the prison population: increase in the diagnosis of cases compared to sputum smear microscopy; reduction in diagnosis time, initiating treatment and isolation; identification of strains resistant to antibiotic therapy; reducing the prevalence and occurrence of tuberculosis; high agreement of test results with culture results; lower cost of the test when carried out in groups of samples or when screening is carried out by radiography. Conclusion: the literature indicated that the rapid molecular test is relevant for combating tuberculosis in prison units, so its use should be considered by authorities and managers as a strategic tool for controlling the disease.

RESUMEN Objetivo: mapear las repercusiones del uso de la prueba molecular rápida para el diagnóstico de tuberculosis en personas privadas de libertad en la literatura científica. Método: scoping review, siguiendo las recomendaciones del Joanna Briggs Institute y PRISMA for Scoping Reviews. La búsqueda se realizó utilizando vocabularios controlados y libres en las siguientes bases de datos: EMBASE, Scopus, MEDLINE, Cinahl, Academic Search Premier, LILACS y Web of Science, en la Biblioteca Digital Brasileña de Tesis y Disertaciones y en Google Scholar. Los materiales que respondieron a la pregunta de revisión fueron seleccionados por dos revisores independientes, basándose en la lectura de títulos, resúmenes y publicaciones. Se incluyeron todo tipo de estudios y publicaciones. Los datos extraídos fueron sometidos a síntesis narrativa y presentados gráficamente. Resultados: entre las 461 publicaciones encontradas, 13 fueron incluidas en la revisión. Los estudios señalaron las siguientes repercusiones del uso de la prueba molecular rápida en la población penitenciaria: aumento del diagnóstico de casos en comparación con la baciloscopia de esputo; reducción del tiempo de diagnóstico, inicio de tratamiento y aislamiento; identificación de cepas resistentes a la terapia con antibióticos; reducir la prevalencia y aparición de la tuberculosis; alta concordancia de los resultados de las pruebas con los resultados del cultivo; menor coste de la prueba cuando se realiza en grupos de muestras o cuando el cribado se realiza mediante radiografía. Conclusión: la literatura indicó que la prueba molecular rápida es relevante para el combate a la tuberculosis en las unidades penitenciarias, por lo que su uso debe ser considerado por autoridades y gestores como una herramienta estratégica para el control de la enfermedad.

RESUMO Objetivo: mapear as repercussões da utilização do teste rápido molecular para o diagnóstico de tuberculose entre as pessoas privadas de liberdade junto à literatura científica. Método: revisão de escopo seguiram-se as recomendações do Joanna Briggs Institute e do PRISMA for Scoping Reviews. A busca foi realizada com vocabulários controlados e livres nas bases de dados: EMBASE, Scopus, MEDLINE, Cinahl, Academic Search Premier, LILACS e Web of Science, na Biblioteca Digital Brasileira de Teses e Dissertações e no Google Scholar. Foram selecionados por dois revisores independentes, os materiais que respondiam à pergunta da revisão, a partir da leitura dos títulos, resumos e publicações. Foram incluídos todos os tipos de estudo e publicações. Os dados extraídos foram submetidos à síntese narrativa e apresentados graficamente. Resultados: entre as 461 publicações encontradas, 13 foram incluídas na revisão. Os estudos apontaram as seguintes repercussões da utilização do teste rápido molecular na população prisional: aumento no diagnóstico de casos comparado à baciloscopia; redução no tempo de diagnóstico, início do tratamento e isolamento; identificação de cepas resistentes à antibioticoterapia; redução da prevalência e ocorrência da tuberculose; alta concordância dos resultados do teste com os da cultura; menor custo do teste quando realizado em grupos de amostras ou quando o rastreamento é realizado por radiografia. Conclusão: a literatura apontou que o teste rápido molecular é relevante para o enfrentamento da tuberculose nas unidades prisionais, de modo que a sua utilização deve ser considerada pelas autoridades e gestores como uma ferramenta estratégica para o controle da doença.

Características de la Diabetes del adulto de inicio juvenil y su diagnóstico en el laboratorio

La Diabetes del adulto de inicio juvenil, es un subtipo hereditario poco común que se manifiesta a una edad temprana, relacionado con mutaciones en genes específicos que principalmente afectan la función de las células beta pancreática. Un diagnóstico preciso es fundamental para un tratamiento efectivo, aunque puede ser desafiante debido a la variabilidad en sus características clínicas y moleculares. Esta revisión analiza la evidencia disponible sobre estas características y los métodos de diagnóstico utilizados en laboratorio. Se realizó una búsqueda exhaustiva en bases de datos científicas, seleccionando estudios relevantes según criterios específicos. Se analizaron características clínicas, hallazgos moleculares y métodos de diagnóstico, utilizando tablas, gráficos y síntesis narrativas. Se identificaron mutaciones genéticas asociadas con MODY, así como biomarcadores útiles en el laboratorio clínico. Además, se describieron métodos de diagnóstico molecular, incluyendo la secuenciación de próxima generación (NGS). Esta revisión resalta la importancia del diagnóstico preciso de MODY, subrayando la diversidad de sus características biológicas y moleculares, y la necesidad de una investigación más profunda para mejorar su identificación y manejo clínico.

Maturity Onset Diabetes of the Young is a rare hereditary subtype that manifests at an early age, related to mutations in specific genes that primarily affect the function of pancreatic beta cells. An accurate diagnosis is crucial for effective treatment, though it can be challenging due to variability in clinical and molecular characteristics. This review examines available evidence on these characteristics and laboratory diagnostic methods. A comprehensive search was conducted in scientific databases, selecting relevant studies based on specific criteria. Clinical features, molecular findings, and diagnostic methods were analyzed using tables, graphs, and narrative synthesis. Genetic mutations associated with MODY were identified, as well as useful biomarkers in clinical laboratory settings. Additionally, molecular diagnostic methods were described, including next-generation sequencing (NGS). This review emphasizes the importance of precise MODY diagnosis, highlighting the diversity of its biological and molecular characteristics, and the need for further research to enhance its identification and clinical management.

A diabetes adulto de início juvenil é um subtipo hereditário raro que se manifesta em uma idade precoce, relacionado a mutações em genes específicos que afetam principalmente a função das células beta do pâncreas. Um diagnóstico preciso é fundamental para um tratamento eficaz, embora possa ser desafiador devido à variabilidade em suas características clínicas e moleculares. Esta revisão analisa a evidência disponível sobre essas características e os métodos de diagnóstico utilizados em laboratório. Foi realizada uma busca abrangente em bases de dados científicas, selecionando estudos relevantes com base em critérios específicos. Características clínicas, descobertas moleculares e métodos de diagnóstico foram analisados utilizando tabelas, gráficos e síntese narrativa. Foram identificadas mutações genéticas associadas ao MODY, assim como biomarcadores úteis em laboratório clínico. Além disso, foram descritos métodos de diagnóstico molecular, incluindo a sequenciação de próxima geração (NGS). Esta revisão enfatiza a importância do diagnóstico preciso do MODY, destacando a diversidade de suas características biológicas e moleculares e a necessidade de uma pesquisa mais aprofundada para melhorar sua identificação e manejo clínico.

Rede nacional de testes moleculares para detecção de Chlamydia trachomatis e Neisseria gonorrhoeae: experiência de implantação-piloto no Brasil / A national network of molecular tests to detect Chlamydia trachomatis and Neisseria gonorrhoeae: a pilot implementation experience in Brazil / Red nacional de pruebas moleculares para la detección de Chlamydia trachomatis y Neisseria gonorrhoeae: experiencia de implantación piloto en Brasil

Resumo: O objetivo deste estudo foi conhecer a opinião dos profissionais participantes da implantação-piloto de testes moleculares para detecção de Chlamydia trachomatis e Neisseria gonorrhoeae no Sistema Único de Saúde (SUS). Determinou-se a taxa de detecção de C. trachomatis e/ou N. gonorrhoeae e os fatores associados à infecção. A estratégia contou com laboratórios pertencentes à rede de carga viral de HIV e hepatites virais. A testagem teve como público-alvo pessoas mais vulnerabilizadas às infecções sexualmente transmissíveis, com coleta de amostras de urina e/ou swabs vaginal, endocervical e/ou uretral masculino. Questionários foram enviados aos gestores estaduais e profissionais de laboratório sobre a implantação-piloto. De maneira geral, as avaliações foram positivas. Entre as fraquezas, citou-se dificuldades na mudança do processo de trabalho, carência de recursos humanos, pouca sensibilidade de profissionais da assistência e ausência de tubo primário de urina, único insumo não fornecido. Como fortaleza, destaca-se aquisição centralizada de testes, compartilhamento de equipamentos e armazenamento de amostras à temperatura ambiente. Das 16.177 pessoas testadas, 1.004 (6,21%) foram positivas para C. trachomatis, 1.036 (6,4%) para N. gonorrhoeae e 239 (1,48%) para C. trachomatis/N. gonorrhoeae. A detecção de infecção ocorreu mais em pessoas jovens (≤ 24 vs. > 24 anos) (aOR = 2,65; IC95%: 2,38-2,96), do sexo masculino (aOR = 1,95; IC95%: 1,72-2,21), pardas/pretas (aOR = 1,06; IC95%: 1,05-1,11), na Região Sudeste (aOR = 1,08; IC95%: 1,02-1,13) e em amostras de secreção uretral (aOR = 1,46; IC95%: 1,41-1,52). Os resultados deste estudo demonstraram a importância da disponibilização da testagem em âmbito nacional, os quais subsidiaram a implantação da rede definitiva para detecção de C. trachomatis/N. gonorrhoeae no SUS.

Abstract: This study aimed to know the opinion of professionals participating in an experiment to implement a pilot for molecular tests to detect Chlamydia trachomatis and Neisseria gonorrhoeae at the Brazilian Unified National Health System (SUS). The detection rate of C. trachomatis and/or N. gonorrhoeae and the factors associated with infection were determined. The strategy included laboratories belonging to the HIV and viral hepatitis viral load network. Testing targeted people who are more vulnerable to sexually transmitted infections and collected urine samples and/or vaginal, endocervical, and/or male urethral swabs. Questionnaires were sent to state managers and laboratory professionals about the implementation of the pilot. Reviews were overall positive. Weaknesses included difficulties changing work processes, lack of human resources, poorly sensitized care professionals, and absence of primary urine tubes, the only input not provided. Strengths included the centralized acquisition of tests, sharing of equipment, and storage of samples at room temperature. Of the 16,177 people who were tested, 1,004 (6.21%) were positive for C. trachomatis; 1,036 (6.4%), for N. gonorrhoeae; and 239 (1.48%), for C. trachomatis/N. gonorrhoeae . Detection of any infection occurred more frequently in young people (≤ 24 vs. > 24 years) (adjOR = 2.65; 95%CI: 2.38-2.96), men (adjOR = 1.95; 95%CI: 1.72-2.21), brown/black individuals (adjOR = 1.06; 95%CI: 1.05-1.11), those in Southeastern Brazil (adjOR = 1.08; 95%CI: 1.02-1.13), and in urethral secretion samples (adjOR = 1.46; 95%CI: 1.41-1.52). Results show the importance of making testing available nationwide, which supported the implementation of a definitive network to detection C. trachomatis/N. gonorrhoeae in SUS.

Resumen: El objetivo de este estudio fue conocer la opinión de los profesionales participantes de la implantación piloto de pruebas moleculares para la detección de Chlamydia trachomatis y Neisseria gonorrhoeae en el Sistema Único de Salud brasileño (SUS). Se determinó la tasa de detección de C. trachomatis y/o N. gonorrhoeae y los factores asociados con la infección. En la estrategia participaron laboratorios pertenecientes a la red de carga viral de VIH y hepatitis virales. La prueba tuvo como público objetivo a personas más vulnerables a las infecciones de transmisión sexual, con recolección de muestras de orina y/o swabs vaginal, endocervicales y/o uretral masculino. Se enviaron cuestionarios a los gestores estatales y a los profesionales de laboratorio sobre la implementación piloto. En general, las evaluaciones fueron positivas. Entre las debilidades, se citó las dificultades en el cambio del proceso de trabajo, la falta de recursos humanos, los profesionales de la asistencia poco sensibilizados y la ausencia del contenedor de orina primaria, el único insumo no suministrado. Como fortalezas, se destaca la adquisición centralizada de pruebas, el intercambio de equipos y el almacenamiento de muestras a temperatura ambiente. De las 16.177 personas evaluadas, 1.004 (6,21%) fueron positivas para C. trachomatis, 1.036 (6,4%) para N. gonorrhoeae y 239 (1,48%) para C. trachomatis/N. gonorrhoeae. La detección de alguna infección ocurrió más en personas jóvenes (≤ 24 vs. > 24 años) (aOR = 2,65; IC95%: 2,38-2,96), del sexo masculino (aOR = 1,95; IC95%: 1,72-2,21), parda/negra (aOR = 1,06; IC95%: 1,05-1,11), localizadas en la región Sudeste (aOR = 1,08; IC95%: 1,02-1,13) y en muestras de secreción uretral (aOR = 1,46; IC95%: 1,41-1,52). Los resultados de este estudio demostraron la importancia de la disponibilidad de la prueba a nivel nacional, los cuales subsidiaron la implantación de la red definitiva para detección de C. trachomatis/N. gonorrhoeae en el SUS.

Controles de calidad externos en el diagnóstico molecular de enfermedades genéticas. Impacto sobre la calidad de los estudios genéticos / External quality controls in the molecular diagnosis of genetic diseases. Impact on the quality of the genetic studies / Controles de qualidade externos no diagnóstico molecular de doenças genéticas. Impacto na qualidade dos estudos genéticos

Resumen La participación en programas de evaluación externa de la calidad (PEEC) dirigidos al diagnóstico de enfermedades genéticas permite obtener una medida objetiva del desempeño técnico y analítico de los laboratorios y es un requisito para la acreditación de los laboratorios clínicos bajo la norma ISO 15189. El objetivo de este estudio fue evaluar retrospectivamente el desempeño en los esquemas EMQN (European Molecular Genetics Quality Network) y CF Network (Cystic Fibrosis European Network) en el período 2014-2022. Se participó en un total de 88 esquemas. Se recolectó la información de nuestros puntajes y las medias de los laboratorios participantes en las categorías genotipificación, interpretación y exactitud de la información del paciente/informe. Se informó en forma completa el 90,9% (n=80) de los esquemas. El desempeño en genotipificación mostró puntajes superiores a la media en el 89,3% de los esquemas; 0,8% de los informes correspondieron a falsos negativos. En interpretación, el 66,7% de los esquemas evidenció un desempeño superior a la media y el 33,3% debajo de la media. La exactitud de la información del paciente/informe presentó puntajes superiores a la media en el 97,6% de los esquemas. Se observó una diferencia estadísticamente significativa en el porcentaje de esquemas con puntaje por encima de la media en el año 2022 (10/12 esquemas) respecto al año 2014 (1/6 esquemas) en la categoría interpretación (p=0,0128). En conclusión, la participación regular en PEEC tuvo impacto positivo en la calidad de los estudios y permite realizar mejoras continuas a partir de las recomendaciones sugeridas por estos programas.

Abstract Participation in external quality assessment programmes focused on rare genetic diseases makes it possible to assess the laboratory technical and analytical performance and it is a prerequisite for accreditation according to ISO 15189. The objective of this study was to perform a retrospective evaluation of our performance in the EMQN (European Molecular Genetics Quality Network) and the CF Network (Cystic Fibrosis European Network) programmes in the 2014-2022 period. The laboratory performance on genotyping, interpretation and clerical accuracy and patient identifiers in a total of 88 schemes were assessed. The information of our scores and the mean scores of all participating laboratories in the three categories were collected. A total of 90.9% of the schemes were fully completed. The performance in genotyping showed scores above the mean scores in 89.3% of the schemes; 0.8% of the reports correspond to false negative results. Regarding interpretation category, 66.7% of the schemes presented scores above the mean scores and 33.3% below the mean scores. The clerical accuracy and patient identifiers were above the mean scores in 97.6% of the schemes. A statistically significant difference in the percentage of schemes with a score above the mean for the interpretation category in the year 2022 (10/12 schemes) was observed compared to the year 2014 (1/6 schemes) (p=0.0128). In conclusion, regular participation in external quality assessment programmes had a positive impact on the quality of the studies and allows for continuous improvements based on the recommendations suggested by these programmes.

Resumo A participação em programas de avaliação externa da qualidade (PEECs) voltados para o diagnóstico de doenças genéticas permite obter uma mensuração objetiva do desempenho técnico e analítico dos laboratórios e é requisito para a acreditação dos laboratórios clínicos sob a norma ISO 15189. O objetivo desse estudo foi avaliar retrospectivamente o desempenho nos esquemas EMQN (European Molecular Genetics Quality Network) e CF Network (Cystic Fibrosis European Network) no período 2014-2022. Participou-se em um total de 88 esquemas. Foram coletadas informações de nossos escores e das médias dos laboratórios participantes nas categorias genotipagem, interpretação e precisão da informação do paciente/laudo. 90,9% (n=80) dos esquemas foram informados em sua totalidade. O desempenho na genotipagem apresentou escores acima da média em 89,3% dos esquemas; 0,8% dos laudos corresponderam a falsos negativos. Na interpretação, 66,7% dos esquemas apresentaram desempenho acima da média e 33,3% abaixo da média. A precisão das informações do paciente/laudo apresentou escores acima da média em 97,6% dos esquemas. Observou-se diferença estatisticamente significativa no percentual de esquemas com pontuação acima da média no ano de 2022 (10/12 esquemas) em relação ao ano de 2014 (1/6 esquemas) na categoria interpretação (p=0,0128). Em conclusão, a participação regular em PEECs teve um impacto positivo na qualidade dos estudos e permite fazer melhorias contínuas com base nas recomendações sugeridas por esses programas.