RESUMEN
This study aimed to standardise an in-house real-time polymerase chain reaction (rtPCR) to allow quantification of hepatitis B virus (HBV) DNA in serum or plasma samples, and to compare this method with two commercial assays, the Cobas Amplicor HBV monitor and the Cobas AmpliPrep/Cobas TaqMan HBV test. Samples from 397 patients from the state of São Paulo were analysed by all three methods. Fifty-two samples were from patients who were human immunodeficiency virus and hepatitis C virus positive, but HBV negative. Genotypes were characterised, and the viral load was measure in each sample. The in-house rtPCR showed an excellent success rate compared with commercial tests; inter-assay and intra-assay coefficients correlated with commercial tests (r = 0.96 and r = 0.913, p < 0.001) and the in-house test showed no genotype-dependent differences in detection and quantification rates. The in-house assay tested in this study could be used for screening and quantifying HBV DNA in order to monitor patients during therapy.
Asunto(s)
ADN Viral/aislamiento & purificación , Técnicas de Genotipaje/normas , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B Crónica/diagnóstico , Técnicas de Diagnóstico Molecular , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Cartilla de ADN/normas , Estudios de Evaluación como Asunto , Femenino , Genotipo , Seropositividad para VIH/sangre , Seropositividad para VIH/diagnóstico , Virus de la Hepatitis B/genética , Hepatitis B Crónica/sangre , Hepatitis C/sangre , Hepatitis C/diagnóstico , Humanos , Lactante , Invenciones/normas , Masculino , Persona de Mediana Edad , Técnicas de Diagnóstico Molecular/instrumentación , Técnicas de Diagnóstico Molecular/métodos , Sensibilidad y Especificidad , Carga Viral , Adulto JovenRESUMEN
INTRODUCTION: HBV and HIV have identical transmission routes. The aim of this study was to determine the prevalence of HBV in HIV patients and to detect the presence of occult HBV infection. METHODS: All samples were tested for serology markers and using qPCR. RESULTS: This study included 232 individuals, out of which 36.6% presented with HBV markers and 11.8% presented with HBsAg or HBV-DNA, including 3 patients that showed OBI. CONCLUSIONS: We observed a high prevalence of HBV among HIV patients. In addition, the results suggest that OBI can occur in patients with serological profiles that are indicative of past infection. Therefore, the application of molecular tests may enable the identification of infections that are not evident solely based on serology.
Asunto(s)
Infecciones por VIH/epidemiología , Anticuerpos contra la Hepatitis B/sangre , Antígenos del Núcleo de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/sangre , Virus de la Hepatitis B/inmunología , Hepatitis B/epidemiología , Brasil/epidemiología , ADN Viral/sangre , Infecciones por VIH/complicaciones , Hepatitis B/complicaciones , Hepatitis B/diagnóstico , Humanos , Prevalencia , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
INTRODUCTION: Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) have identical transmission routes, explaining the high prevalence of coinfections. The main aim of this study was to detect fluctuations in serological HCV levels in HIV patients. METHODS: We analyzed samples of 147 patients who attended an outpatient service that supports HIV/AIDS patients in São Paulo city. We also recruited 22 HCV-monoinfected patients who attended the Instituto Adolfo Lutz Laboratory in São Paulo city, to compare the test results. Serological testing of the blood samples was performed for the detection of HCV antibodies. The samples were then analyzed using real-time PCR for RNA viral quantification and sequencing. RESULTS: We found that 13.6% of the study population was coinfected with HIV and HCV. In 20% of coinfected patients, fluctuations in serology results were detected in samples collected during the follow-up. No changes in anti-HCV serological markers were observed in HCV-monoinfected patients. An HCV viral load was detected in 9,5% of the samples collected from HIV patients. CONCLUSIONS: Our findings provide important clinical data to public health professionals and highlight the importance of periodic monitoring of HCV/HIV coinfected patients.
Asunto(s)
Infecciones por VIH/complicaciones , Anticuerpos contra la Hepatitis C/sangre , Hepatitis C/complicaciones , ARN Viral/sangre , Recuento de Linfocito CD4 , Coinfección , Femenino , Genotipo , Hepacivirus/genética , Hepacivirus/inmunología , Hepatitis C/diagnóstico , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena en Tiempo Real de la Polimerasa , Carga ViralRESUMEN
Abstract INTRODUCTION: HBV and HIV have identical transmission routes. The aim of this study was to determine the prevalence of HBV in HIV patients and to detect the presence of occult HBV infection. METHODS: All samples were tested for serology markers and using qPCR. RESULTS: This study included 232 individuals, out of which 36.6% presented with HBV markers and 11.8% presented with HBsAg or HBV-DNA, including 3 patients that showed OBI. CONCLUSIONS: We observed a high prevalence of HBV among HIV patients. In addition, the results suggest that OBI can occur in patients with serological profiles that are indicative of past infection. Therefore, the application of molecular tests may enable the identification of infections that are not evident solely based on serology.
Asunto(s)
Humanos , Infecciones por VIH/epidemiología , Virus de la Hepatitis B/inmunología , Hepatitis B/epidemiología , Anticuerpos contra la Hepatitis B/sangre , Antígenos del Núcleo de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/sangre , Brasil/epidemiología , ADN Viral/sangre , Infecciones por VIH/complicaciones , Prevalencia , Reacción en Cadena en Tiempo Real de la Polimerasa , Hepatitis B/complicaciones , Hepatitis B/diagnósticoRESUMEN
INTRODUCTION: This study aimed to monitor the seasonality of rotavirus infection, and gain insight into the variability of Brazilian strains. METHODS: A total of 28 stool samples were analyzed from 698 revised cases of gastroenteritis during a norovirus outbreak in the summer of 2010 in Guarujá, Brazil. Diagnosis was performed using enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and sequencing. RESULTS: Rotavirus infection was detected in 17.9% (5/28) of samples; 4 samples were G2P[4] genotype, and one G2P[4]+P[6] genotype. G2 and P[4] sequences showed a genetic relationship to strains from India and Russia, respectively. CONCLUSIONS: The seasonal pattern of rotavirus may be a consequence of human activity apart from climate factors.
Asunto(s)
Infecciones por Caliciviridae/epidemiología , Brotes de Enfermedades , Gastroenteritis/epidemiología , Norovirus/aislamiento & purificación , Infecciones por Rotavirus/epidemiología , Rotavirus/aislamiento & purificación , Adolescente , Brasil/epidemiología , Ensayo de Inmunoadsorción Enzimática , Heces/virología , Femenino , Gastroenteritis/virología , Genotipo , Humanos , Masculino , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rotavirus/genética , Infecciones por Rotavirus/diagnóstico , Estaciones del AñoRESUMEN
Abstract INTRODUCTION: Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) have identical transmission routes, explaining the high prevalence of coinfections. The main aim of this study was to detect fluctuations in serological HCV levels in HIV patients. METHODS: We analyzed samples of 147 patients who attended an outpatient service that supports HIV/AIDS patients in São Paulo city. We also recruited 22 HCV-monoinfected patients who attended the Instituto Adolfo Lutz Laboratory in São Paulo city, to compare the test results. Serological testing of the blood samples was performed for the detection of HCV antibodies. The samples were then analyzed using real-time PCR for RNA viral quantification and sequencing. RESULTS We found that 13.6% of the study population was coinfected with HIV and HCV. In 20% of coinfected patients, fluctuations in serology results were detected in samples collected during the follow-up. No changes in anti-HCV serological markers were observed in HCV-monoinfected patients. An HCV viral load was detected in 9,5% of the samples collected from HIV patients. CONCLUSIONS: Our findings provide important clinical data to public health professionals and highlight the importance of periodic monitoring of HCV/HIV coinfected patients.
Asunto(s)
Humanos , Masculino , Femenino , ARN Viral/sangre , Infecciones por VIH/complicaciones , Hepatitis C/complicaciones , Anticuerpos contra la Hepatitis C/sangre , Hepatitis C/diagnóstico , Hepacivirus/genética , Hepacivirus/inmunología , Recuento de Linfocito CD4 , Carga Viral , Coinfección , Reacción en Cadena en Tiempo Real de la Polimerasa , Genotipo , Persona de Mediana EdadRESUMEN
This study aimed to standardise an in-house real-time polymerase chain reaction (rtPCR) to allow quantification of hepatitis B virus (HBV) DNA in serum or plasma samples, and to compare this method with two commercial assays, the Cobas Amplicor HBV monitor and the Cobas AmpliPrep/Cobas TaqMan HBV test. Samples from 397 patients from the state of São Paulo were analysed by all three methods. Fifty-two samples were from patients who were human immunodeficiency virus and hepatitis C virus positive, but HBV negative. Genotypes were characterised, and the viral load was measure in each sample. The in-house rtPCR showed an excellent success rate compared with commercial tests; inter-assay and intra-assay coefficients correlated with commercial tests (r = 0.96 and r = 0.913, p < 0.001) and the in-house test showed no genotype-dependent differences in detection and quantification rates. The in-house assay tested in this study could be used for screening and quantifying HBV DNA in order to monitor patients during therapy.
Asunto(s)
Humanos , Masculino , Femenino , Lactante , Preescolar , Niño , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Adulto Joven , ADN Viral/aislamiento & purificación , Técnicas de Genotipaje/normas , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B Crónica/diagnóstico , Técnicas de Diagnóstico Molecular , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Cartilla de ADN/normas , Estudios de Evaluación como Asunto , Genotipo , Seropositividad para VIH/sangre , Seropositividad para VIH/diagnóstico , Virus de la Hepatitis B/genética , Hepatitis B Crónica/sangre , Hepatitis C/sangre , Hepatitis C/diagnóstico , Invenciones/normas , Técnicas de Diagnóstico Molecular/instrumentación , Técnicas de Diagnóstico Molecular/métodos , Sensibilidad y Especificidad , Carga ViralRESUMEN
The aim of this study was to evaluate the sensitivity and specificity of the Ridascreen(®) Norovirus 3rd Generation kit compared to the RT-PCR. A retrospective, descriptive study was conducted with 245 specimens from sporadic cases and outbreak surveillance samples of gastroenteritis in Brazil from 2006 to 2009. Overall, the kit showed a sensitivity of 61.8% and a specificity of 92.5%. The sensitivity for outbreaks diagnosis was 87.9% and specificity 83.8%. The Ridascreen(®) 3rd Generation could detect specimen containing genogroup (G) II with high sensitivity. However, GI and mixed infections (GI/GII) were unlikely to be detected by the kit. ELISA for Norovirus (NoV) detection provides a rapid, technically simple assay system that can be used to increase the surveillance of gastroenteritis outbreaks, especially in Public Health Laboratories with high sample throughput. This assay is useful for the detection of NoV outbreaks and is an improvement as compared to previous ELISAs; however, due to its sensitivity, RT-PCR in still required for routine NoV detection in sporadic cases investigation.
Asunto(s)
Infecciones por Caliciviridae/diagnóstico , Técnicas de Laboratorio Clínico/métodos , Gastroenteritis/virología , Norovirus/aislamiento & purificación , Brasil , Infecciones por Caliciviridae/virología , Humanos , Técnicas de Diagnóstico Molecular/métodos , Epidemiología Molecular/métodos , Juego de Reactivos para Diagnóstico , Sensibilidad y EspecificidadRESUMEN
Objective: the aim of this study was to identify HBV genotypes in serum samples from patients from the state of São Paulo, received by the viral hepatitis laboratory, at the Virology Centre of Instituto Adolfo Lutz, from various municipalities. Methods: a total of 94 serum samples were randomly analyzed. Genotyping was performed using nested PCR for amplification of S and Pol regions from viral genome. Genotypes were identified comparing the sequences obtained with the sequences deposited in GenBank. Results: we were able to determine the genotype of 91 (97%) samples, as follows: genotype A (55.3%), D (32%), F (5.3%), C (3.2%) and G (1%). There are few data on the epidemiology of genotype G. This genotype has been detected in restricted areas around the world. Frequently, the genotype G infection occurs in HIV-positive male patients. In our case, the sample identified as G was also positive for HIV but in a female patient, which is an uncommon finding in the scientific literature. Conclusion: in this work, we identified the most frequent genotypes in São Paulo as well as the genotype G, rare among the genotypes found in our environment. .
Objetivo: o objetivo deste estudo foi identificar os genótipos do HBV nas amostras de soros recebidas pelo Laboratório de Hepatites do Centro de Virologia do Instituto Adolfo Lutz. Métodos: foram analisadas aleatoriamente 94 amostras de soropositivas, provenientes de diversos municípios do Estado de São Paulo. Para determinação dos genótipos, foi realizada Nested-PCR das regiões S e Pol do HBV. Os genótipos foram identificados comparando os resultados amplificados com as sequências depositadas no GenBank. Resultados: foi possível determinar o genótipo de 91 (97%) amostras do total analisado e os genótipos identificados foram: genótipos A (55,3%), D (32%), F (5,3%), C (3,2%) e G (1%). Há poucos dados a respeito da epidemiologia do genótipo G. Esse genótipo tem sido detectado em áreas restritas do mundo. Geralmente, a infecção pelo genótipo G ocorre em indivíduos HIV positivos do sexo masculino. Neste trabalho, a amostra identificada como G foi também positiva para HIV e era de uma paciente do sexo feminino, dado raro na literatura científica. Conclusão: neste trabalho, foram identificados os genótipos mais frequentes, assim como uma cepa do genótipo G, rara entre os encontrados em nosso meio. .
RESUMEN
Introduction This study aimed to monitor the seasonality of rotavirus infection, and gain insight into the variability of Brazilian strains. Methods A total of 28 stool samples were analyzed from 698 revised cases of gastroenteritis during a norovirus outbreak in the summer of 2010 in Guarujá, Brazil. Diagnosis was performed using enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and sequencing. Results Rotavirus infection was detected in 17.9% (5/28) of samples; 4 samples were G2P[4] genotype, and one G2P[4]+P[6] genotype. G2 and P[4] sequences showed a genetic relationship to strains from India and Russia, respectively. Conclusions The seasonal pattern of rotavirus may be a consequence of human activity apart from climate factors. .
Asunto(s)
Adolescente , Femenino , Humanos , Masculino , Infecciones por Caliciviridae/epidemiología , Brotes de Enfermedades , Gastroenteritis/epidemiología , Norovirus/aislamiento & purificación , Infecciones por Rotavirus/epidemiología , Rotavirus/aislamiento & purificación , Brasil/epidemiología , Ensayo de Inmunoadsorción Enzimática , Heces/virología , Genotipo , Gastroenteritis/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Infecciones por Rotavirus/diagnóstico , Rotavirus/genética , Estaciones del AñoRESUMEN
A infecção pelo vírus da hepatite B (HBV) é um problema global de saúde pública e estima-se que mais de dois bilhões de indivíduos estejam infectados por HBV em todo o mundo. Entre eles, aproximadamente 240 milhões estão cronicamente infectados e 780 mil vêm a óbito em consequência desta infecção. O diagnóstico laboratorial da hepatite B é realizado através da pesquisa de antígenos e anticorpos no soro/plasma. A hepatite B oculta é definida pela presença de HBV-DNA na ausência de antígeno de superfície (S), com ou sem anticorpos detectáveis...
Hepatitis B virus (HBV) infection is a global health problem. It is estimated that two billion people are infected with HBV around the world. Among them, it is possible that chronic infection affects 240 million people and 780,000 die due this infection. Hepatitis B laboratory diagnosis is accomplished by antigens and antibodies research in serum/plasma. Occult hepatites B is defined like the presence of HBV DNA in the serum/plasma of individuals testing HBsAg negative, with detectable or undetectable antibodies...