RESUMO
Introduction: Despite the availability of an effective vaccine and treatment to reduce the viral load and progressive hepatocellular injury, approximately 240 million people worldwide are chronically infected with the hepatitis B virus (HBV). In Colombia, the circulation of different viral genotypes has been confirmed. Mutations in the genome have been associated to antiviral therapy resistance, viral escape to neutralizing antibodies, occult infection and progression to hepatocellular carcinoma. Objective: To identify the genotypes and the presence of mutations in the coding region of the surface (S) antigen and the reverse transcriptase (RT) domain of the polymerase of HBV obtained from serum samples for hepatitis B diagnosis received by the Instituto Nacional de Salud during the period 2002-2014. Materials and methods: A total of 495 serum samples with previous HBsAg reactive result were used for molecular detection. A fragment of 1,591 nucleotides was sequenced, and the corresponding phylogenetic analysis was performed. Results: We detected the viral genome of HBV in 66 samples and 28 were successfully sequenced. The phylogenetic analysis allowed the identification of subgenotypes F3 and A2. The L180M and M204V resistance mutations were simultaneously identified in one sample, while the I169L resistance mutation was identified in another one. A single escape mutation, P120Q, was identified in one more. Two samples showed a deletion of 105 nucleotides in the preS1-preS2 region. Conclusions: The circulation of genotypes/subgenotypes F3 and A2 of HBV in Colombia was corroborated, as well as the presence of some resistance and escape mutations. The present study constitutes a contribution to the molecular epidemiology of HBV in Colombia.
Assuntos
Genes Virais , Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/genética , DNA Polimerase Dirigida por RNA/genética , Proteínas Virais/genética , Sequência de Bases , DNA Viral/sangue , DNA Viral/genética , Farmacorresistência Viral/genética , Variação Genética/genética , Genótipo , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/enzimologia , Humanos , Mutação de Sentido Incorreto , Filogenia , Mutação Puntual , Domínios Proteicos , DNA Polimerase Dirigida por RNA/sangue , Estudos Retrospectivos , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Proteínas Virais/sangueRESUMO
Introducción. Se estima que 240 millones de personas en el mundo tienen infección crónica con el virus de la hepatitis B (HBV). En Colombia, la endemia es variable y circulan diferentes genotipos virales. Las mutaciones a lo largo del genoma se han asociado con resistencia antiviral, el escape ante la reacción de anticuerpos neutralizadores tras la vacunación o a la infección natural, la infección oculta y la progresión a carcinoma hepatocelular. Objetivo. Identificar los genotipos y las mutaciones presentes en la región codificante del antígeno de superficie (S) y del dominio de la transcriptasa inversa (reverse transcriptase, RT) de la polimerasa del HBV en muestras de suero remitidas al Instituto Nacional de Salud de Colombia para el diagnóstico de hepatitis B, entre el 2002 y el 2014. Materiales y métodos. En 495 muestras de suero positivas para el antígeno de superficie de la hepatitis B (HBsAg) se buscó el ADN viral, se amplificó y secuenció un fragmento de 1.591 nucleótidos y, posteriormente, se hizo el análisis filogenético correspondiente. Resultados. En 66 de las muestras se logró detectar el genoma viral y 28 de ellas se secuenciaron exitosamente. El análisis filogenético permitió identificar los genotipos y subgenotipos F3 y A2. Una muestra presentó simultáneamente las sustituciones de resistencia L180M y M204V, otra presentó la sustitución I169L y en una se identificó la mutación P120Q, previamente asociada con variantes de escape. Dos muestras presentaron una deleción de 105 nucleótidos en la región preS1-preS2. Conclusiones. Se corroboró la circulación en Colombia de los genotipos y subgenotipos F3 y A2, así como la presencia de mutaciones de resistencia y escape. El presente estudio constituye un aporte a la epidemiologia molecular del HBV en Colombia.
Introduction: Despite the availability of an effective vaccine and treatment to reduce the viral load and progressive hepatocellular injury, approximately 240 million people worldwide are chronically infected with the hepatitis B virus (HBV). In Colombia, the circulation of different viral genotypes has been confirmed. Mutations in the genome have been associated to antiviral therapy resistance, viral escape to neutralizing antibodies, occult infection and progression to hepatocellular carcinoma. Objective: To identify the genotypes and the presence of mutations in the coding region of the surface (S) antigen and the reverse transcriptase (RT) domain of the polymerase of HBV obtained from serum samples for hepatitis B diagnosis received by the Instituto Nacional de Salud during the period 2002-2014. Materials and methods: A total of 495 serum samples with previous HBsAg reactive result were used for molecular detection. A fragment of 1,591 nucleotides was sequenced, and the corresponding phylogenetic analysis was performed. Results: We detected the viral genome of HBV in 66 samples and 28 were successfully sequenced. The phylogenetic analysis allowed the identification of subgenotypes F3 and A2. The L180M and M204V resistance mutations were simultaneously identified in one sample, while the I169L resistance mutation was identified in another one. A single escape mutation, P120Q, was identified in one more. Two samples showed a deletion of 105 nucleotides in the preS1-preS2 region. Conclusions: The circulation of genotypes/subgenotypes F3 and A2 of HBV in Colombia was corroborated, as well as the presence of some resistance and escape mutations. The present study constitutes a contribution to the molecular epidemiology of HBV in Colombia.
Assuntos
Vírus da Hepatite B , Genótipo , DNA Polimerase Dirigida por RNA , MutaçãoRESUMO
Introducción. El virus de la hepatitis E se ha convertido en un problema de salud pública, especialmente en los países en desarrollo. Se conocen cuatro genotipos en mamíferos, de los cuales el G3 se ha encontrado en hepatitis autóctonas en países y regiones con gran población de cerdos, y el G1 se ha asociado a muertes maternas. Objetivo. Determinar la infección simultánea con el virus de la hepatitis E y sus genotipos circulantes en Colombia en 1.097 sueros utilizando los marcadores serológicos de los virus de las hepatitis A, B y C. Materiales y métodos. Se seleccionaron 1.097 sueros provenientes de diferentes municipios de Colombia, conservados en el Laboratorio de Virología del Instituto Nacional de Salud. Se determinaron los anticuerpos IgG e IgM anti-hepatitis E. A los positivos se les amplificó el genoma viral mediante reacción en cadena de la polimerasa convencional. Los productos se secuenciaron y analizaron filogenéticamente y se los comparó con las secuencias del ORF2 registradas en el GenBank. Resultados. Se identificaron 278 sueros positivos para IgG anti-hepatitis E, 62 para IgM y 64 para ambos marcadores. La infección simultánea con los virus de la hepatitis E y la hepatitis A determinada por IgG anti-hepatitis E fue de 33,6 % y por IgM anti-hepatitis E fue de 16,1 %; la infección simultánea por los virus de la hepatitis E y B fue de 23,4 % y 8,1 %, y por los virus de la hepatitis E y C fue de 35,4 % y 5,83 %, respectivamente. De las 52 muestras positivas en la reacción en cadena de la polimerasa convencional, nueve secuencias se agruparon como genotipo 3a de origen porcino, cepa norteamericana. Conclusiones. La mayor seropositividad se registró para las hepatitis A y E. La frecuencia de la infección simultánea con el virus de la hepatitis E y otros virus hepatótropos indica que este patógeno puede ser más frecuente de lo esperado. La circulación del genotipo 3a implica que esta enfermedad puede presentarse en forma de brote y de zoonosis en Colombia.
Introduction: Hepatitis E virus has emerged as a public health problem, particularly in developing countries. The four genotypes identified in mammals include the G3 found in indigenous hepatitis in countries and regions with high porcine population, and the G1, associated with maternal deaths. Objective: To determine coinfection by hepatitis E virus and the circulating genotypes in Colombia in 1,097 samples using serological markers for hepatitis A, B and C. Materials and methods: Serum samples of 1,097 patients from different regions of Colombia stored at the Laboratorio de Virología of the Instituto Nacional de Salud were selected to detect IgG and IgM anti-hepatitis E virus antibodies. The viral genomes of positive samples were amplified by RT-PCR, and the products were sequenced and phylogenetically analyzed by comparing ORF2 sequences deposited in the GenBank. Results: IgG anti-hepatitis E virus antibodies were found in 278 samples, IgM in 62, and both markers in 64. Hepatitis E virus and hepatitis A virus coinfection determined by IgG anti-hepatitis E virus was 33.6% and 16.1% by IgM; hepatitis E virus and hepatitis B virus coinfection was 23.4% and 8.1%, and hepatitis E virus and hepatitis C virus coinfection was 35.4% and 5.83%, respectively. Among the 52 positive samples by PCR nine were sequenced and grouped within genotype 3A of the American porcine strain. Conclusions: The highest seropositivity was observed for hepatitis A and E. The incidence of hepatitis E virus coinfection with other hepatotropic viruses indicated that this pathogen is more frequent than expected. The circulation of genotype 3A implies that this disease may occur in outbreaks and as zoonosis in Colombia.
Assuntos
Coinfecção , Vírus da Hepatite E , Genótipo , Hepatite ARESUMO
INTRODUCTION: Hepatitis E virus has emerged as a public health problem, particularly in developing countries. The four genotypes identified in mammals include the G3 found in indigenous hepatitis in countries and regions with high porcine population, and the G1, associated with maternal deaths. OBJECTIVE: To determine coinfection by hepatitis E virus and the circulating genotypes in Colombia in 1,097 samples using serological markers for hepatitis A, B and C. MATERIALS AND METHODS: Serum samples of 1,097 patients from different regions of Colombia stored at the Laboratorio de Virología of the Instituto Nacional de Salud were selected to detect IgG and IgM anti-hepatitis E virus antibodies. The viral genomes of positive samples were amplified by RT-PCR, and the products were sequenced and phylogenetically analyzed by comparing ORF2 sequences deposited in the GenBank. RESULTS: IgG anti-hepatitis E virus antibodies were found in 278 samples, IgM in 62, and both markers in 64. Hepatitis E virus and hepatitis A virus coinfection determined by IgG anti-hepatitis E virus was 33.6% and 16.1% by IgM; hepatitis E virus and hepatitis B virus coinfection was 23.4% and 8.1%, and hepatitis E virus and hepatitis C virus coinfection was 35.4% and 5.83%, respectively. Among the 52 positive samples by PCR nine were sequenced and grouped within genotype 3A of the American porcine strain. CONCLUSIONS: The highest seropositivity was observed for hepatitis A and E. The incidence of hepatitis E virus coinfection with other hepatotropic viruses indicated that this pathogen is more frequent than expected. The circulation of genotype 3A implies that this disease may occur in outbreaks and as zoonosis in Colombia.
