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2.
Med. infant ; 28(1): 23-26, Marzo 2021. ilus, Tab
Article in Spanish | LILACS, BINACIS, UNISALUD | ID: biblio-1282888

ABSTRACT

Pneumocystis jirovecii es un hongo oportunista, causante de neumonía en huéspedes inmunocomprometidos. Es una infección grave con elevada tasa de mortalidad en pacientes oncohematológicos y receptores de trasplante de células progenitoras hematopoyéticas. La administración de corticosteroides es el principal factor de riesgo para adquirir esta infección. Actualmente las infecciones ocurren en aquellos pacientes que no reciben adecuada profilaxis. Las técnicas de diagnóstico molecular son las recomendadas por su elevada sensibilidad, especificidad y rapidez. La frecuencia global de P. jirovecii en pacientes inmunocomprometidos de nuestro hospital, durante el período evaluado fue de 4,8%, con una mortalidad global del 20%. Como factores de mal pronóstico se reportan la presencia de coinfecciones y la necesidad de asistencia respiratoria mecánica. Es importante la sospecha precoz en pacientes de riesgo, confirmada con un diagnóstico preciso mediante métodos moleculares para una intervención adecuada y oportuna (AU)


Pneumocystis jirovecii is an opportunistic fungus, causing pneumonia in immunocompromised hosts. It is a severe infection with a high mortality rate in oncology/hematology patients and hematopoietic stem cell transplant recipients. The administration of corticosteroids is the main risk factor for acquiring this infection. Currently infections occur in patients who do not receive adequate prophylaxis. Molecular diagnostic techniques are recommended because of their high sensitivity, specificity, and speed. In the study period, the overall incidence of P. jirovecii in immunocompromised patients at our hospital was 4.8%, with an overall mortality rate of 20%. Factors of a poor prognosis are the presence of coinfections and the need for mechanical respiratory assistance. Early suspicion in high-risk patients is important to confirm the diagnosis through molecular studies and start adequate and early treatment (AU)


Subject(s)
Humans , Infant , Child, Preschool , Child , Adolescent , Polymerase Chain Reaction/methods , Pneumocystis Infections/diagnosis , Pneumocystis Infections/epidemiology , Immunocompromised Host , Molecular Diagnostic Techniques/methods , Pneumocystis carinii/isolation & purification , Hospitals, Pediatric/statistics & numerical data , Cross-Sectional Studies , Retrospective Studies
3.
Braz. arch. biol. technol ; 64: e21190423, 2021. tab, graf
Article in English | LILACS | ID: biblio-1285548

ABSTRACT

Abstract High sensitivity of qPCR assay can be compromised by the presence of PCR inhibitors in samples analyzed. The aim of this study was to analyze the RT-qPCR assay efficiency considering the RNA quality/quantity and the presence of PCR inhibitors in patients with chemotherapy and/or antibiotic therapy. We analyzed 60 samples using RT-qPCR from individuals suspected of leukemia and 44 samples were quantified by fluorimetry and spectrophotometry. The efficiency of the RT-qPCR assay was evaluated comparing the threshold cycle (Ct) from tested samples and the standard curve. The 260/280 and 260/230 ratios, the presence of PCR inhibitors and the amount of sample (ng) used in the RT-qPCR reaction can be associated with 56.8% (R²=0.56, p<0.05) in the Ct obtained. The decrease of the RT-qPCR efficiency can be explained in 42,8% due to the variation of the 260/280 ratio (R²=0.42,p<0.05). The presence of antibiotics in the blood sample can be associated in 11.3% with the variability of 260/280 ratio (R²=0.11,p<0.05). Presence of chemotherapeutic drugs in the blood sample was not correlated with Ct variation (p=0.17). The spectrophotometer determines a RNA quantification with 2.2 times higher than the fluorimeter (t=2.2, p=0,03) and this difference is correlated with the 260/280 ratio (R²=0.36, p<0.05). Samples with low purity had a reduction in the qPCR efficiency, although we did not observe false results.


Subject(s)
Humans , Polymerase Chain Reaction/methods , Nucleic Acid Synthesis Inhibitors , Molecular Diagnostic Techniques/methods , Spectrophotometry/instrumentation , Fluorometry/instrumentation
4.
Article in English | WPRIM | ID: wpr-887737

ABSTRACT

Objective@#To evaluate multidrug resistant loop-mediated isothermal amplification (MDR-LAMP) assay for the early diagnosis of multidrug-resistant tuberculosis and to compare the mutation patterns associated with the @*Methods@#MDR-LAMP assay was evaluated using 100 @*Results@#The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of MDR-LAMP were 85.5%, 93.6%, 96.7%, and 74.4% for the detection of resistance to isoniazid and rifampicin, respectively, and 80.5%, 92.3%, 98.6%, and 41.4% for the detection of @*Conclusion@#MDR-LAMP is a rapid and accessible assay for the laboratory identification of rifampicin and isoniazid resistance of


Subject(s)
Antitubercular Agents , Bacterial Proteins/genetics , Catalase/genetics , DNA, Bacterial/analysis , DNA-Directed RNA Polymerases/genetics , Drug Resistance, Multiple, Bacterial/genetics , Isoniazid , Molecular Diagnostic Techniques/methods , Mutation , Mycobacterium tuberculosis/isolation & purification , Nucleic Acid Amplification Techniques/methods , Oxidoreductases/genetics , Phenotype , Rifampin , Whole Genome Sequencing
5.
Rev. cuba. med. trop ; 72(3): e532, sept.-dic. 2020. tab
Article in Spanish | LILACS, CUMED | ID: biblio-1156544

ABSTRACT

Introducción: En la actualidad las infecciones fúngicas representan un problema para la salud humana. Las infecciones causadas por especies patógenas de hongos registran un incremento constante y se ubican entre el cuarto y décimo lugar como causa de muerte, particularmente en las unidades de cuidado intensivo. Un diagnóstico adecuado y precoz impacta directamente en la morbilidad y mortalidad asociadas a estas. Objetivo: Describir las principales técnicas de diagnóstico no convencional de las enfermedades fúngicas más frecuentes, en especial las relacionadas con el diagnóstico serológico y molecular. Métodos: Se realizó una revisión de la literatura científica sobre el tema, publicada entre 2000 y 2019. Se revisaron un total de 63 trabajos. Como motores de búsqueda se emplearon Google y Google Scholar. Se revisaron las bases de datos Medline, PubMed, Science Direct, BUCea y SciELO. Análisis y síntesis de la información: Las técnicas serológicas se emplean en el diagnóstico de las micosis invasivas o sistémicas por ser fáciles, rápidas y confiables. La detección de anticuerpos tiene utilidad limitada en el diagnóstico de las micosis invasivas debido a que la respuesta puede estar retrasada, reducida o no existir en pacientes inmunocomprometidos. La detección de componentes no antigénicos liberados por los hongos durante la infección y la secuenciación de ácidos nucleicos fúngicos son otras opciones para el diagnóstico de las micosis. Conclusiones: El desarrollo biotecnológico aporta nuevas herramientas que incrementan las oportunidades de identificación de las micosis. En la actualidad se disponen de métodos basados tanto en la detección de marcadores inmunológicos como de elementos moleculares específicos. La eficacia de las herramientas no convencionales para el diagnóstico depende de la correcta combinación de estas(AU)


Introduction: Fungal infections are a current human health problem. Infections caused by pathogenic fungal species constantly increase in number, and are ranked between the fourth and tenth leading causes of death, particularly in intensive care units. Early accurate diagnosis has a direct impact on the morbidity and mortality of fungal infections. Objective: Describe the main non-conventional diagnostic techniques for the most common fungal diseases, especially those related to serological and molecular diagnosis. Methods: A review was conducted of the scientific literature about the topic published between the years 2000 and 2019. A total 63 publications were reviewed. The search engines used were Google and Google Scholar. The databases Medline, PubMed, Science Direct, BUCea and SciELO were reviewed. Data analysis and synthesis: Serological techniques are used for the diagnosis of invasive or systemic mycoses because they are easy, fast and reliable. The detection of antibodies has a limited usefulness in invasive mycosis diagnosis, for the response may be delayed, reduced or inexistent in immunocompromised patients. Detection of non-antigenic components released by fungi during infection and sequencing of fungal nucleic acids are other mycosis diagnosis options. Conclusions: Biotechnological development contributes new tools increasing mycosis identification opportunities. Methods are currently available which are based on detection of immunological markers and specific molecular elements. The efficacy of non-conventional diagnostic tools depends on their appropriate combination(AU)


Subject(s)
Humans , Molecular Diagnostic Techniques/methods , Mycoses/diagnosis , Mycoses/mortality , Clinical Laboratory Techniques/methods
6.
Rev. colomb. cancerol ; 24(3): 140-145, jul.-set. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1144333

ABSTRACT

Resumen El desarrollo y la innovación de nuevas tecnologías ha permitido mejorar la detección de la infección por el virus del papiloma humano de alto riesgo. La captura de híbridos II es un ensayo que se basa en hibridación y quimioluminiscencia. Cobas VPH Test es una PCR cualitativa y Aptima VPH Assay permite detectar la expresión de ARN mensajero de las oncoproteínas E6/E7 del VPH de alto riesgo. Estas técnicas presentan ventajas en comparación con la citología convencional, que se utiliza como prueba de rutina para la detección temprana del cáncer de cuello uterino. En el estudio ESTAMPA se realizaron 13.691 procesamientos que permitieron identificar que para el planteamiento de proyectos de investigación o para la implementación de pruebas de tamizaje de VPH es necesario analizar las ventajas y desventajas de las pruebas del mercado.


Abstract The development and innovation of new technologies has improved the detection of high-risk human papillomavirus infection. Hybrid capture II is an assay that is based on hybridization and chemiluminescence. Cobas HPV Test is a qualitative PCR and Aptima HPV Assay allows to detect the expression of messenger RNA of the high- risk HPV E6 / E7 oncoproteins. These techniques have advantages, in comparison, with conventional cytology that is routinely used for the detection of cervical cancer. In the ESTAMPA study, 13,691 prosecutions were carried out that allowed to identify that for the planning of research projects or for the implementation of HPV screening tests, it is necessary to analyze the advantages and disadvantages of market tests.


Subject(s)
Humans , Female , Papillomaviridae/isolation & purification , Research Design , Uterine Cervical Neoplasms/diagnosis , Papillomavirus Infections/diagnosis , Molecular Diagnostic Techniques/methods , Papillomaviridae/genetics , DNA, Viral , RNA, Messenger , Uterine Cervical Neoplasms/genetics , Mass Screening , Multicenter Studies as Topic , Triage , Papillomavirus Infections/genetics , Human Papillomavirus DNA Tests , Luminescent Measurements , Nucleic Acid Hybridization
7.
Afr. j. lab. med. (Online) ; 8(1): 1-6, 2019. tab
Article in English | AIM, AIM | ID: biblio-1257327

ABSTRACT

Background: The Presto combined qualitative real-time assay for Chlamydia trachomatis and Neisseria gonorrhoeae (Presto CT/NG PCR assay) is appealing for developing countries, because it can be used with multiple DNA extraction methods and polymerase chain reaction (PCR) platforms.Objectives: The objective of the study was to implement and evaluate the Presto CT/NG PCR assay at the National Reference Laboratory (NRL) in Kigali, Rwanda, where no real-time PCR assays for the detection of C. trachomatis or N. gonorrhoeae were available.Methods: The Presto CT/NG PCR assay was first evaluated at the Institute of Tropical Medicine (ITM) in Antwerp, Belgium. Next, NRL laboratory technicians were trained to use the assay on their ABI PRISM 7500 real-time PCR instrument and their competencies were assessed prior to trial initiation. During the trial, endocervical swabs were tested at the NRL, with bi-monthly external quality control testing monitored by the ITM. The final NRL results were evaluated against extended gold standard testing at the ITM, consisting of the Abbott m2000 RealTime System with confirmation of positive results by an in-house real-time PCR assay for C. trachomatis or N. gonorrhoeae.Results: Of the 192 samples analysed using the Presto assay at the NRL, 16 samples tested positive for C. trachomatis and 17 tested positive for N. gonorrhoeae; four of these were infected with both. The sensitivity and specificity of the Presto assay were 93.3% (95% confidence interval [CI]: 68.1% ­ 99.8%) and 99.4% (95% CI: 96.8% ­ 100%) for C. trachomatis and 100% (95% CI: 76.8% ­ 100%) and 98.8% (95% CI: 95.8% ­ 99.9%) for N. gonorrhoeae.Conclusion: C. trachomatis and N. gonorrhoeae testing with the Presto assay was feasible in Kigali, Rwanda, and good performance was achieved


Subject(s)
Biological Assay , Chlamydia trachomatis , Molecular Diagnostic Techniques/methods , Neisseria gonorrhoeae
8.
J. bras. pneumol ; 45(2): e20180185, 2019. tab
Article in English | LILACS | ID: biblio-1002431

ABSTRACT

ABSTRACT Objective: To evaluate the accuracy of rapid molecular testing as a diagnostic tool and estimate the incidence of smear-positive pulmonary tuberculosis among the indigenous population. Methods: This is an epidemiological study based on secondary data. We calculated the incidence of smear-positive pulmonary tuberculosis between January 1st, 2011 and December 31, 2016, and the performance of bacilloscopy and rapid molecular testing in diagnosing pulmonary tuberculosis compared to sputum culture (standard test). Results: We included 4,048 cases of indigenous people with respiratory symptoms who provided sputum samples for analysis. Among them, 3.7%, 6.7%, and 3.7% had positive results for bacilloscopy, sputum culture, and rapid molecular testing, respectively. The mean incidence of pulmonary tuberculosis was 269.3/100 thousand inhabitants. Rapid molecular testing had 93.1% sensitivity and 98.2% specificity, compared to sputum culture. Bacilloscopy showed 55.1% sensitivity and 99.6% specificity. Conclusions: Rapid molecular testing can be useful in remote areas with limited resources and a high incidence of tuberculosis, such as indigenous villages in rural regions of Brazil. In addition, the main advantages of rapid molecular testing are its easy handling, fast results, and the possibility of detecting rifampicin resistance. Together, these attributes enable the early start of treatment, contributing to reduce the transmission in communities recognized as vulnerable to infection and disease.


RESUMO Objetivo: Avaliar a acurácia do teste rápido molecular como ferramenta diagnóstica e estimar a incidência de casos pulmonares positivos entre a população indígena. Métodos: Estudo epidemiológico baseado em dados secundários. Foi calculada a incidência de casos de tuberculose pulmonar positiva entre 1° de janeiro de 2011 e 31 de dezembro de 2016, e o desempenho da baciloscopia e do teste rápido molecular no diagnóstico de tuberculose pulmonar, em comparação à cultura de escarro (teste padrão). Resultados: Foram incluídos 4.048 casos de indígenas considerados sintomáticos respiratórios, que forneceram amostras de escarro para análise. Destes, 3,7%, 6,7% e 3,7% apresentaram resultados positivos para baciloscopia, cultura e teste rápido molecular, respectivamente. A incidência média de tuberculose pulmonar foi de 269,3/100 mil habitantes. A sensibilidade do teste rápido molecular, em relação à cultura, foi 93,1% e a especificidade foi 98,2%. A baciloscopia apresentou sensibilidade 55,1% e especificidade 99,6%. Conclusões: O teste rápido molecular pode ser útil em áreas remotas, com recursos limitados e incidência de tuberculose elevada, como as aldeias indígenas nas áreas rurais do país. Ademais, o teste rápido molecular apresenta como principais vantagens o fácil manuseio, os resultados rápidos e a possibilidade de identificar a resistência à rifampicina. Em conjunto, esses atributos facilitam o início do tratamento precoce, contribuindo para reduzir a transmissão em comunidades reconhecidamente vulneráveis à infecção e à doença.


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/ethnology , Indians, South American/statistics & numerical data , Molecular Diagnostic Techniques/methods , Mycobacterium leprae/isolation & purification , Reference Values , Sputum/microbiology , Time Factors , Tuberculosis, Pulmonary/microbiology , Brazil/epidemiology , Incidence , Cross-Sectional Studies , Reproducibility of Results , Sensitivity and Specificity , Sex Distribution , Age Distribution
9.
J. bras. pneumol ; 45(2): e20180128, 2019. tab, graf
Article in English | LILACS | ID: biblio-1002440

ABSTRACT

ABSTRACT Objective: To evaluate the rapid diagnosis of multidrug-resistant tuberculosis, by using a commercial line probe assay for rifampicin and isoniazid detection (LPA-plus), in the routine workflow of a tuberculosis reference laboratory. Methods: The LPA-plus was prospectively evaluated on 341 isolates concurrently submitted to the automated liquid drug susceptibility testing system. Results: Among 303 phenotypically valid results, none was genotypically rifampicin false-susceptible (13/13; 100% sensitivity). Two rifampicin-susceptible isolates harboured rpoB mutations (288/290; 99.3% specificity) which, however, were non-resistance-conferring mutations. LPA-plus missed three isoniazid-resistant isolates (23/26; 88.5% sensitivity) and detected all isoniazid-susceptible isolates (277/277; 100% specificity). Among the 38 (11%) invalid phenotypic results, LPA-plus identified 31 rifampicin- and isoniazid-susceptible isolates, one isoniazid-resistant and six as non-Mycobacterium tuberculosis complex. Conclusions: LPA-plus showed excellent agreement (≥91%) and accuracy (≥99%). Implementing LPA-plus in our setting can speed up the diagnosis of multidrug-resistant tuberculosis, yield a significantly higher number of valid results than phenotypic drug susceptibility testing and provide further information on the drug-resistance level.


RESUMO Objetivo: Avaliar o diagnóstico rápido de tuberculose multirresistente, utilizando um teste comercial de sondas em linha (LPA-plus), na rotina de um laboratório de referência de tuberculose. Métodos: O teste LPA-plus foi avaliado prospectivamente em 341 isolados simultaneamente submetidos ao teste de suscetibilidade aos antimicrobianos em meio líquido, pelo sistema automatizado. Resultados: Entre os 303 resultados fenotipicamente válidos, nenhum foi genotipicamente falso suscetível à rifampicina (13/13; 100% de sensibilidade). Dois isolados sensíveis à rifampicina apresentavam mutações no gene rpoB (288/290; especificidade de 99,3%), as quais, no entanto, não são associadas à resistência a rifampicina. O LPA-plus não identificou resistência à isoniazida em três isolados fenotipicamente resistentes (23/26; 88,5% de sensibilidade) e detectou todos os isolados sensíveis à isoniazida (277/277; especificidade de 100%). Entre os 38 (11%) resultados fenotípicos inválidos, o LPA-plus identificou 31 isolados sensíveis à rifampicina e à isoniazida, um resistente à isoniazida e seis como micobactérias não tuberculosas. Conclusões: O LPA-plus mostrou excelente concordância (≥91%) e acurácia (≥99%). Sua implementação pode acelerar o diagnóstico da tuberculose multirresistente, produzir número significativamente maior de resultados válidos do que o teste fenotípico de suscetibilidade aos antimicrobianos e fornecer informações adicionais sobre o nível de resistência aos fármacos.


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Tuberculosis, Multidrug-Resistant/diagnosis , Molecular Diagnostic Techniques/methods , Phenotype , Rifampin/pharmacology , Time Factors , DNA, Bacterial , Microbial Sensitivity Tests , Prospective Studies , Reproducibility of Results , Sensitivity and Specificity , Tuberculosis, Multidrug-Resistant/microbiology , Nucleic Acid Amplification Techniques/methods , Early Diagnosis , Isoniazid/pharmacology , Mycobacterium tuberculosis/isolation & purification , Mycobacterium tuberculosis/drug effects , Antitubercular Agents/pharmacology
10.
J. bras. pneumol ; 45(2): e20170451, 2019. tab
Article in English | LILACS | ID: biblio-1040271

ABSTRACT

ABSTRACT Tuberculosis continues to be a major public health problem worldwide. The aim of the present study was to evaluate the accuracy of the Xpert MTB/RIF rapid molecular test for tuberculosis, using pulmonary samples obtained from patients treated at the Júlia Kubitschek Hospital, which is operated by the Hospital Foundation of the State of Minas Gerais, in the city of Belo Horizonte, Brazil. This was a retrospective study comparing the Xpert MTB/RIF test results with those of standard culture for Mycobacterium tuberculosis and phenotypic susceptibility tests. Although the Xpert MTB/RIF test showed high accuracy for the detection of M. tuberculosis and its resistance to rifampin, attention must be given to the clinical status of the patient, in relation to the test results, as well as to the limitations of molecular tests.


RESUMO A tuberculose permanece como um grave problema de saúde pública. O objetivo deste estudo foi avaliar a acurácia do teste rápido molecular Xpert MTB/RIF em amostras pulmonares no Hospital Júlia Kubitschek, Fundação Hospitalar do Estado de Minas Gerais, localizado em Belo Horizonte (MG). Trata-se de um estudo descritivo retrospectivo, considerando-se como método padrão a cultura para o bacilo da tuberculose e o teste de sensibilidade fenotípico. O teste Xpert MTB/RIF apresentou ótima acurácia para a detecção da tuberculose e resistência à rifampicina, mas é necessária a atenção a dados clínicos do paciente em relação ao resultado do exame e às limitações dos testes moleculares.


Subject(s)
Humans , Sputum/microbiology , Trachea/microbiology , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/microbiology , Bronchoalveolar Lavage Fluid/microbiology , Nucleic Acid Amplification Techniques/methods , Rifampin/pharmacology , DNA, Bacterial , Reproducibility of Results , Retrospective Studies , Sensitivity and Specificity , Molecular Diagnostic Techniques/methods , Drug Resistance, Bacterial/drug effects , Tertiary Care Centers , Antibiotics, Antitubercular/pharmacology , Mycobacterium tuberculosis/isolation & purification , Mycobacterium tuberculosis/drug effects
11.
Arch. endocrinol. metab. (Online) ; 62(6): 576-584, Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-983811

ABSTRACT

ABSTRACT Approximately 15-30% of all thyroid nodules evaluated with fine-needle aspiration biopsy (FNAB) are classified as cytologically indeterminate. The stepwise unraveling of the molecular etiology of thyroid nodules has provided the basis for a better understanding of indeterminate samples and an opportunity to decrease diagnostic surgery in this group of patients. Over the last 15 years, several studies have tested different methodologies to detect somatic mutations (by polymerase chain reaction and next-generation sequencing, for example), and to identify differentially expressed genes or microRNA, aiming at developing molecular tests to improve the presurgical diagnosis of cytologically indeterminate nodules. In this review, we will provide an overview of the currently available molecular tests and the impact of mutation testing on the diagnosis of thyroid cancer. We will also review current published data and future perspectives in molecular testing of thyroid nodule FNAB and describe the current Brazilian experience with this diagnostic approach. Based on currently available data, especially for countries outside the US-Europe axis, a rational use of these tests must be made to avoid errors with regard to test indication and interpretation of test outcomes. In addition to clinical, radiological, and cytological features, we still need to determine local malignancy rates and conduct more independent validation and comparative performance studies of these tests before including them into our routine approach to indeterminate FNAB.


Subject(s)
Humans , Thyroid Nodule/diagnosis , Thyroid Nodule/pathology , Molecular Diagnostic Techniques/standards , Biopsy, Fine-Needle , Mutation , Brazil , Sensitivity and Specificity , Thyroid Nodule/genetics , Molecular Diagnostic Techniques/methods
12.
Braz. j. microbiol ; 49(2): 422-428, Apr.-June 2018. tab, graf
Article in English | LILACS | ID: biblio-889236

ABSTRACT

Abstract Identification of nonfermenting Gram-negative bacteria (NFGNB) of cystic fibrosis patients is hard and misidentification could affect clinical outcome. This study aimed to propose a scheme using polymerase chain reaction to identify NFGNB. This scheme leads to reliable identification within 3 days in an economically viable manner when compared to other methods.


Subject(s)
Humans , Polymerase Chain Reaction/methods , Gram-Negative Bacterial Infections/diagnosis , Cystic Fibrosis/complications , Molecular Diagnostic Techniques/methods , Gram-Negative Bacteria/isolation & purification , Time Factors , Gram-Negative Bacteria/genetics
13.
J. bras. pneumol ; 44(2): 112-117, Mar.-Apr. 2018. tab, graf
Article in English | LILACS | ID: biblio-893903

ABSTRACT

ABSTRACT Objective: To evaluate the impact of the use of the molecular test for Mycobacterium tuberculosis and its resistance to rifampin (Xpert MTB/RIF), under routine conditions, at a referral hospital in the Brazilian state of Bahia. Methods: This was a descriptive study using the database of the Mycobacteriology Laboratory of the Octávio Mangabeira Specialized Hospital, in the city of Salvador, and georeferencing software. We evaluated 3,877 sputum samples collected from symptomatic respiratory patients, under routine conditions, between June of 2014 and March of 2015. All of the samples were submitted to sputum smear microscopy and the Xpert MTB/RIF test. Patients were stratified by gender, age, and geolocation. Results: Among the 3,877 sputum samples evaluated, the Xpert MTB/RIF test detected M. tuberculosis in 678 (17.5%), of which 60 (8.8%) showed resistance to rifampin. The Xpert MTB/RIF test detected M. tuberculosis in 254 patients who tested negative for sputum smear microscopy, thus increasing the diagnostic power by 59.9%. Conclusions: The use of the Xpert MTB/RIF test, under routine conditions, significantly increased the detection of cases of tuberculosis among sputum smear-negative patients.


RESUMO Objetivo: Avaliar o impacto do teste rápido molecular automatizado Xpert MTB/RIF, utilizado para a detecção de Mycobacterium tuberculosis e sua resistência à rifampicina, em condições de rotina, em um hospital de referência no estado da Bahia. Métodos: Estudo descritivo retrospectivo utilizando o banco de dados do Laboratório de Micobacteriologia do Hospital Especializado Octávio Mangabeira, localizado na cidade de Salvador, e um programa de georreferenciamento. Entre junho de 2014 e março de 2015, foram incluídas no estudo 3.877 amostras de escarro coletadas de pacientes sintomáticos respiratórios em condições de rotina. Todas as amostras coletadas foram submetidas tanto à baciloscopia quanto a Xpert MTB/RIF. Os pacientes foram estratificados por sexo, idade e georreferenciamento. Resultados: Das 3.877 amostras de escarro analisadas, Xpert MTB/RIF detectou a presença de M. tuberculosis em 678 pacientes (17,5%). Desses, 60 (8,8%) apresentaram resistência à rifampicina. O Xpert MTB/RIF detectou 254 pacientes com baciloscopia negativa, representando um acréscimo diagnóstico de 59,9%. Conclusões: A implantação do Xpert MTB/RIF, sob condições de rotina, teve um impacto significativo no aumento da detecção de casos de tuberculose em pacientes com baciloscopia negativa.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Sputum/microbiology , Tuberculosis/diagnosis , Molecular Diagnostic Techniques/methods , Diagnostic Tests, Routine/methods , Mycobacterium tuberculosis/isolation & purification , Reference Values , Rifampin/therapeutic use , Tuberculosis/microbiology , Tuberculosis/drug therapy , Brazil , Reproducibility of Results , Retrospective Studies , Sensitivity and Specificity , Treatment Outcome , Drug Resistance, Bacterial/drug effects , Tertiary Care Centers , Microscopy/methods , Antibiotics, Antitubercular/therapeutic use , Mycobacterium tuberculosis/drug effects
14.
Arq. Inst. Biol ; 85: e0842016, 2018. ilus, tab
Article in English | LILACS, VETINDEX | ID: biblio-996678

ABSTRACT

In areas where human tuberculosis and bovine tuberculosis coexist, differentiation between M. bovis and M. tuberculosis is important for monitoring the spread of M. bovis among cattle and from cattle to humans. The objective of this study was to isolate and identify M. bovis in bovines with positive diagnosis identified on tuberculin test in the State of Paraíba, Northeastern Brazil. Thirty-two bovines that tested positive in the comparative tuberculin test were used, from which samples of any organ with lesions suggestive of tuberculosis were collected, as well as lymph nodes, when no gross lesions were observed. Samples were submitted to histopathological exam, mycobacterial culture, Ziehl-Neelsen staining and molecular diagnosis. Twenty-one (65.6%) animals presented lesions suggestive of tuberculosis. As to body region 77.7% of lesions were found in the thoracic cavity, 12.4% in the head and 9.9% in the abdominal cavity. Among 55 samples submitted to mycobacterial culture, mycobacteria were isolated in 31 (56.4%), being 13 (41.9%) identified as M. bovis and 18 (58.1%) as Mycobacterium spp. Conclusion is that isolation and identification of M. bovis and Mycobacterium spp. in cattle suggests that humans are exposed to the risk of infection. This reinforces the need for intensification and optimization of prevention and control measures foreseen in the Brazilian National Program for the Control and Eradication of Bovine Brucellosis and Tuberculosis. Mycobacteria isolation and identification surveys are, therefore, encouraged in other Northeastern states.(AU)


Em áreas onde a tuberculose humana e a tuberculose bovina coexistem, a diferenciação entre M. bovis e M. tuberculosis é importante para monitorar a disseminação de M. bovis entre bovinos e destes para os seres humanos. Objetivou-se neste estudo isolar e identificar M. bovis em bovinos com diagnóstico positivo pelo teste de tuberculinização no estado da Paraíba, nordeste do Brasil. Foram submetidos 32 bovinos positivos ao teste de tuberculinização comparativa, dos quais foram colhidas amostras de qualquer órgão com lesões sugestivas de tuberculose, e, nos casos em que não foram observadas lesões sugestivas, foram colhidas amostras de linfonodos. As amostras foram submetidas a exame histopatológico, cultivo micobacteriológico, coloração de Ziehl-Neelsen e diagnóstico molecular. Apresentaram lesões sugestivas de tuberculose 21 animais (65,6%). Com relação à distribuição das lesões de acordo com a região corporal, 77,7% localizavam-se na cavidade torácica, 12,4% na cabeça e 9,9% na cavidade abdominal. De 55 amostras submetidas ao cultivo de micobactérias, em 31 (56,4%) foram isoladas micobactérias, sendo que em 13 (41,9%) foi identificado M. bovis, e nas 18 restantes (58,1%) foi identificado Mycobacterium spp. Conclui-se que o isolamento e a identificação de M. bovis e Mycobacterium spp. em bovinos indicam que os seres humanos estão expostos ao risco de infecção. Isso reforça a necessidade de intensificação e otimização de medidas de prevenção e controle previstas no Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose Bovina. Sugere-se a realização de estudos de isolamento e identificação de micobactérias em outros estados do Nordeste.(AU)


Subject(s)
Cattle , Tuberculosis/transmission , Tuberculosis, Bovine/transmission , Immunologic Tests/methods , Brucellosis, Bovine , Molecular Diagnostic Techniques/methods , Mycobacterium
15.
Braz. j. microbiol ; 48(4): 785-790, Oct.-Dec. 2017. tab, graf
Article in English | LILACS | ID: biblio-889167

ABSTRACT

ABSTRACT Early diagnosis of tuberculosis is of major clinical importance. Among 4733 clinical specimens collected from 3363 patients and subjected to Ziehl-Neelsen microscopy, 4109 were inoculated onto Löwenstein-Jensen slants and 3139 in Bactec/9000MB. Polymerase Chain Reaction (PCR) was performed in 3139 specimens, whereas, a genotypic assay was directly applied in 93 Mycobacterium tuberculosis complex PCR-positive for isoniazid and rifampicin resistance detection specimens (GenoType MTBDRplus). Recovered M. tuberculosis isolates (64) as well as, 21 more sent from Regional Hospitals were tested for antimycobacterial resistance with a phenotypic (manual MGIT-SIRE) and a genotypic assay (GenoType MTBDRplus). PCR in the clinical specimens showed excellent specificity (97.4%) and accuracy (96.8%), good sensitivity (70.4%), but low positive predictive value (40.3%). MGIT-SIRE performed to M. tuberculosis did not confer a reliable result in 16 isolates. Of the remaining 69 isolates, 15 were resistant to streptomycin, seven to isoniazid, seven to ethambutol and five to rifampicin. GenoType MTBDRplus correctly detected isoniazid (seven) and rifampicin-resistant M. tuberculosis strains (five), showing an excellent performance overall (100%). Susceptibility results by the molecular assay applied directly to clinical specimens were identical to those obtained from recovered isolates of the corresponding patients. Combining molecular and conventional methods greatly contribute to early diagnosis and accurate susceptibility testing of tuberculosis.


Subject(s)
Humans , Culture Techniques/methods , Molecular Diagnostic Techniques/methods , Mycobacterium tuberculosis/isolation & purification , Tuberculosis, Multidrug-Resistant/diagnosis , Tuberculosis, Pulmonary/diagnosis , Antitubercular Agents/pharmacology , Culture Techniques/economics , Drug Resistance, Bacterial , Genotype , Microbial Sensitivity Tests , Molecular Diagnostic Techniques/economics , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/growth & development , Tuberculosis, Multidrug-Resistant/drug therapy , Tuberculosis, Multidrug-Resistant/microbiology , Tuberculosis, Pulmonary/drug therapy , Tuberculosis, Pulmonary/microbiology
16.
Braz. j. microbiol ; 48(2): 232-236, April.-June 2017. tab
Article in English | LILACS | ID: biblio-839371

ABSTRACT

Abstract Our aim in this study is to compare the standard culture method with the multiplex PCR and the Speed-Oligo® Bacterial Meningitis Test (SO-BMT) – a hybridization-based molecular test method – during the CSF examination of the patients with the pre-diagnosis of acute bacterial meningitis. For the purposes of this study, patients with acute bacterial meningitis treated at the Dicle University Medical Faculty Hospital, Infectious Diseases and Clinical Microbiology Clinic between December 2009 and April 2012 were retrospectively evaluated. The diagnosis of bacterial meningitis was made based on the clinical findings, laboratory test anomalies, CSF analysis results, and the radiological images. Growth was observed in the CSF cultures of 10 out of the 57 patients included in the study (17.5%) and Streptococcus pneumoniae was isolated in all of them. The CSF samples of 34 patients (59.6%) were positive according to the SO-BMT and S. pneumoniae was detected in 33 of the samples (97.05%), while Neisseria meningitidis was found in 1 sample (2.95%). In a total of 10 patients, S. pneumoniae was both isolated in the CSF culture and detected in the SO-BMT. The culture and the SO-BMT were negative in 23 of the CSF samples. There was no sample in which the CSF culture was positive although the SO-BMT was negative. While SO-BMT seems to be a more efficient method than bacterial culturing to determine the pathogens that most commonly cause bacterial meningitis in adults, further studies conducted on larger populations are needed in order to assess its efficiency and uses.


Subject(s)
Streptococcus pneumoniae/isolation & purification , Polymerase Chain Reaction/methods , Bacteriological Techniques/methods , Meningitis, Bacterial/diagnosis , Molecular Diagnostic Techniques/methods , Diagnostic Tests, Routine/methods , Neisseria meningitidis/isolation & purification , Streptococcus pneumoniae/classification , Streptococcus pneumoniae/growth & development , Streptococcus pneumoniae/genetics , Cerebrospinal Fluid/microbiology , Retrospective Studies , Sensitivity and Specificity , Neisseria meningitidis/classification , Neisseria meningitidis/growth & development , Neisseria meningitidis/genetics
17.
Braz. j. microbiol ; 48(2): 366-372, April.-June 2017. tab, graf
Article in English | LILACS | ID: biblio-839381

ABSTRACT

Abstract Malignant Catarrhal Fever (MCF) was investigated in the central nervous system of cattle with neurological syndrome. Two-hundred-ninety samples were analyzed by histology, and molecular methods to detect ovine herpesvirus type 2 (OvHV-2) were optimized and validated. The qualitative polymerase chain reaction (qualitative PCR) analytical sensitivity was 101 DNA copies/µL and found 4.8% (14/290) positive for OvHV-2. The quantitative polymerase chain reaction (qPCR) analytical sensitivity was 100 DNA copy/µL and 5.9% (17/290) positivity, with 47.1% (8/17) of the positive samples presenting histological evidence of non-purulent meningo-encephalitis. The qualitative PCR products (422 bp of the ORF75 region) were sequenced and submitted to phylogenetic analysis. Identity matrices showed 100% similarity in OvHV-2 samples obtained in this study and those recovered from GenBank, corroborating other studies.


Subject(s)
Animals , Phylogeny , Molecular Diagnostic Techniques/methods , Herpesviridae/isolation & purification , Malignant Catarrh/diagnosis , Malignant Catarrh/pathology , Brazil , Cattle , Cluster Analysis , Polymerase Chain Reaction/methods , Sensitivity and Specificity , Sequence Homology , Sequence Analysis, DNA , Genotype , Herpesviridae/classification , Herpesviridae/genetics , Histocytochemistry , Microscopy
18.
Cad. Saúde Pública (Online) ; 33(9): e00214515, 2017. tab, graf
Article in Portuguese | LILACS | ID: biblio-889758

ABSTRACT

Resumo: O objetivo do estudo foi estimar o impacto orçamentário do GeneXpert MTB/RIF para o diagnóstico da tuberculose sob a perspectiva do Programa Nacional de Controle da Tuberculose, valendo-se de um modelo estático apoiado no método epidemiológico entre 2013 e 2017. Comparou-se um teste Xpert MTB/RIF com duas baciloscopias diagnósticas. Utilizaram-se dados epidemiológicos, populacionais, de custos, a taxa de câmbio e bases de dados do Sistema Único de Saúde. Foi realizada análise de sensibilidade por cenários. A incorporação do GeneXpert MTB/RIF demandaria um montante de R$ 147 milhões em cinco anos e representaria um impacto de 23% a 26% nos dois primeiros anos, e de cerca de 11% entre 2015 e 2017. Os resultados podem apoiar os gestores brasileiros e dos países latino-americanos no planejamento e gestão na sua decisão de incorporação da tecnologia.


Abstract: The study aimed to estimate the budget impact of GeneXpert MTB/RIF for diagnosis of tuberculosis from the perspective of the Brazilian National Program for Tuberculosis Control, drawing on a static model using the epidemiological method, from 2013 to 2017. GeneXpert MTB/RIF was compared with two diagnostic sputum smear tests. The study used epidemiological, population, and cost data, exchange rates, and databases from the Brazilian Unified National Health System. Sensitivity analysis of scenarios was performed. Incorporation of GeneXpert MTB/RIF would cost BRL 147 million (roughly USD 45 million) in five years and would have an impact of 23 to 26% in the first two years and some 11% between 2015 and 2017. The results can support Brazilian and other Latin American health administrators in planning and managing the decision on incorporating the technology.


Resumen: El objetivo del estudio fue estimar el impacto presupuestario del GeneXpert MTB/RIF para el diagnóstico de la tuberculosis, desde la perspectiva del Programa Nacional de Control de la Tuberculosis de Brasil, valiéndose de un modelo estático, apoyado en el método epidemiológico entre 2013 y 2017. Se comparó un test Xpert MTB/RIF con dos baciloscopias diagnósticas. Se utilizaron datos epidemiológicos, poblacionales, de costes, la tasa de cambio y bases de datos del Sistema Único de Salud. Se realizó un análisis de sensibilidad por escenarios. La incorporación del GeneXpert MTB/RIF demandaría un montante de R$ 147 millones en cinco años y representaría un impacto de 23 a 26% durante los dos primeros años, y de cerca de un 11% entre 2015 y 2017. Los resultados pueden apoyar a los gestores brasileiros y de los países latinoamericanos en la planificación y gestión a la hora de decidir incorporar este tipo de tecnología.


Subject(s)
Humans , Tuberculosis, Pulmonary/diagnosis , Budgets , Molecular Diagnostic Techniques/economics , Mycobacterium tuberculosis/genetics , Tuberculosis, Pulmonary/economics , Brazil , Sensitivity and Specificity , Molecular Diagnostic Techniques/methods , National Health Programs
20.
Rev. biol. trop ; 64(4): 1505-1518, oct.-dic. 2016. tab, ilus
Article in English | LILACS | ID: biblio-958230

ABSTRACT

Abstract:The productivity of arid legumes, such as Clusterbean (Cyamopsis tetragonoloba), Cowpea (Vigna unguiculata), Moth bean (Vigna aconitifolia) and Horse gram (Macrotyloma uniflorum), may remain stagnant over decades because of their high susceptibility to root diseases. Besides, there is a limitation on the information about molecular diagnosis and intraspecific genetic variability of root pathogens in arid legumes. To contribute in this field, we assessed a total of 52 isolates from 88 root samples that were found infected with fungal pathogens in Jodhpur, Jaipur and Bikaner Districts of Rajasthan. Diseased roots samples were analyzed following standard microbiological methods for fungus extraction and purification, and for genetic studies. Irrespective of the geographical location from where the diseased samples were collected, all pathogen isolates were clustered in RAPD dendrograms as per their respective genera. Phylogram, based on multiple sequence alignment, revealed that different genera (i.e. Fusarium, Neocosmospora and Syncephalastrum), separated from each other, and species within the same genera, clustered together with their reference sequences with apreciable bootstrap values. Out of 20 representative isolates representing each cluster and all outgroups sequenced, eight were molecularly identified as Neocosmospora vasinfecta, five as Fusarium solani, two as Neocosmospora striata, two as Fusarium acutatum, one as Syncephalastrum monosporum, one as Fusarium oxysporum and one as Fusarium species. The root pathogens of the arid legumes were found neither restricted to a geographical location nor were host specific in nature. Fusarium solani wilt in cowpea and seedling rot in moth bean, F. oxysporum wilt in moth bean, F. acutatum damping off in cowpea and Clusterbean, Fusarium sp. seedling rot in Clusterbean, Neocosmospora striata root rot in cowpea and wilt in Clusterbean and Syncephalastrum monosporum root rot in Clusterbean were molecularly identified as new fungal records as pathogens causing root diseases in arid legumes. Rev. Biol. Trop. 64 (4): 1505-1518. Epub 2016 December 01.


Resumen:La producción de leguminosas resistentes a sequías como Cyamopsis tetragonoloba, Vigna unguiculata, Vigna aconitifolia y Macrotyloma uniflorum, puede permanecer inactiva durante décadas debido a su alta susceptibilidad a enfermedades en las raíces. Además, hay información limitada relacionada con el diagnóstico molecular y la variabilidad genética intraespecífica de patógenos de raíces en estas leguminosas resistentes a sequías. Para contribuir en esta área, evaluamos un total de 52 extractos de 88 raíces infectadas con patógenos fúngicos en los distritos de Jodhpur, Jaipur y Bikaner de Rajastán. Las muestras de raíces infectadas se analizaron siguiendo los métodos estándar de microbiología para extracción y purificación de hongos y para estudios genéticos. Independientemente del sitio donde se recolectaron las muestras contaminadas, todos los extractos patógenicos se agruparon en dendrogramas RAPD en cada uno de sus respectivos géneros. El filograma, basado en alineamiento de secuencias múltiples reveló que distintos géneros (Fusarium, Neocosmospora y Syncephalastrum) separados entre ellos y especies del mismo género se agrupan con sus secuencias de referencia con valores de bootstrap significativos. De cada 20 extractos representantes de cada agrupamiento y todos los grupos externos secuenciados, ocho fueron identificados molecularmente como Neocosmospora vasinfecta, dos como Fusarium acutatum, una como Syncephalastrum monosporum, una como Fusarium oxysporum y una como Fusarium. Los patógenos de estas leguminosas resistentes a sequías no están restringidos por la localidad ni por un hospedero específico. Fusarium solani que marchita el frijol de vaca y pudre la semilla de Vigna aconitifolia, F. oxysporum que marchita a Vigna aconitifolia, F. acutatum que marchita a Vigna unguiculata y Cyamopsis tetragonoloba, Fusarium sp. que pudre la semilla de Cyamopsis tetragonoloba, Neocosmospora striata que pudre la raíz de Vigna unguiculata y marchita a Cyamopsis tetragonoloba y, Syncephalastrum monosporum que pudre la raíz en Cyamopsis tetragonoloba, fueron identificados molecularmente como nuevos registros de patógenos fúngicos que causan daños en las raíces de leguminosas resistentes a sequías.


Subject(s)
Plant Diseases/microbiology , Molecular Diagnostic Techniques/methods , Vigna/microbiology , Fusarium/isolation & purification , Hypocreales/isolation & purification , Fabaceae/microbiology , Mucorales/isolation & purification , Genetic Variation , DNA, Fungal , Plant Roots/genetics , Random Amplified Polymorphic DNA Technique , Polymorphism, Single Nucleotide , Vigna/genetics , Hypocreales/genetics , India , Fabaceae/genetics
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