[Reverse hybridization and sequencing for genotyping the hepatitis C virus].
BACKGROUND
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(R) 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.
Levada
Patrícia Martinez
PM
Faculdade de Medicina de Botucatu, Faculdade de Medicina de Botucatu, Universidade Estadual Paulista, Botucatu, SP.
Moraes
Camila Fernanda Verdichio de
CF
Corvino
Silvia Maria
SM
Grotto
Rejane Maria Tommasini
RM
Silva
Giovanni Faria
GF
Pardini
Maria Inês de Moura Campos
MI
D020121:5' Untranslated Regions / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D005838:Genotype; D016174:Hepacivirus / Q000145:classification / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D017361:Viral Nonstructural Proteins / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase
D020121:5' Untranslated Regions; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase