Reverse hybridization and sequencing for genotyping the hepatitis C virus

Revista da Sociedade Brasileira de Medicina Tropical

Moraes, Camila Fernanda Verdichio de¹  Pardini, Maria Inês de Moura Campos¹  Silva, Giovanni Faria¹  Corvino, Silvia Maria¹  Universidade Estadual Paulista, Botucatu¹  Grotto, Rejane Maria Tommasini¹  Levada, Patrícia Martinez¹ 

关键词: Hepatitis C virus;    Genotyping;    Reverse hybridization;    Direct sequencing;   

DOI  :  10.1590/S0037-86822010000200006

学科分类：农业科学（综合）

来源: Sociedade Brasileira de Medicina Tropical

【 摘 要 】

INTRODUCTION: The methods for genotyping the hepatitis C virus have been much discussed. The aim of this study was to compare the methodologies of reverse hybridization and direct sequencing for genotyping the hepatitis C virus.METHODS: Ninety-one plasma samples from patients attended at the Botucatu Medical School, São Paulo State University, were used. Genotyping by reverse hybridization was performed using the INNO-LiPA® v.1.0 commercial kit. Direct sequencing was performed in an automated sequencer using in-house protocols.RESULTS: Genotyping by direct sequencing was shown to be efficient for resolving cases that had remained inconclusive after using the commercial kit. The kit showed erroneous results in relation to virus subtyping. Moreover, direct sequencing revealed an error of the kit regarding the genotypic determination, thereby raising doubts about the efficiency of reverse hybridization for identifying the virus genotype.CONCLUSIONS: Genotyping by direct sequencing allowed greater accuracy of virus classification than did reverse hybridization.

【 授权许可】

Unknown