Genotyping hepatitis C virus from hemodialysis patients in central Brazil by line probe assay and sequence analysis
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Data
2007
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Resumo
The present study examined the distribution of hepatitis C virus
(HCV) genotypes and subtypes in a hemodialysis population in Goiás
State, Central Brazil, and evaluated the efficiency of two genotyping
methods: line probe assay (LiPA) based on the 5' noncoding region
and nucleotide sequencing of the nonstructural 5B (NS5B) region of
the genome. A total of 1095 sera were tested for HCV RNA by RTnested PCR of the 5' noncoding region. The LiPA assay was able to
genotype all 131 HCV RNA-positive samples. Genotypes 1 (92.4%)
and 3 (7.6%) were found. Subtype 1a (65.7%) was the most prevalent,
followed by subtypes 1b (26.7%) and 3a (7.6%). Direct nucleotide
sequencing of 340 bp from the NS5B region was performed in 106
samples. The phylogenetic tree showed that 98 sequences (92.4%)
were classified as genotype 1, subtypes 1a (72.6%) and 1b (19.8%),
and 8 sequences (7.6%) as subtype 3a. The two genotyping methods
gave concordant results within HCV genotypes and subtypes in 100
and 96.2% of cases, respectively. Only four samples presented discrepant results, with LiPA not distinguishing subtypes 1a and 1b.
Therefore, HCV genotype 1 (subtype 1a) is predominant in hemodialysis patients in Central Brazil. By using sequence analysis of the
NS5B region as a reference standard method for HCV genotyping, we
found that LiPA was efficient at the genotype level, although some
discrepant results were observed at the subtype level (sensitivity of
96.1% for subtype 1a and 95.2% for subtype 1b). Thus, analysis of the
NS5B region permitted better discrimination between HCV subtypes,
as required in epidemiological investigations.
Descrição
Palavras-chave
Hepatitis C, Hepatitis C virus, Genotypes, Hemodialysis
Citação
SANTO, M. P. Espírito et al. Genotyping of hepatitis C virus from hemodialysis patients in central Brazil by line probe assay and sequence analysis. Brazilian Journal of Medical and Biological Research, Ribeirão Preto, v. 40, n. 4, p. 545-550, Apr. 2007.