Nested restriction site-specific PCR to detect and type hepatitis C virus (HCV): a rapid method to distinguish HCV subtype 1b from other genotypes

Journal of Clinical Microbiology
L KrekulovaL W Riley

Abstract

Genotypic differentiation of hepatitis C virus (HCV) has become an integral part of clinical management and epidemiologic studies of hepatitis C infections. Thus, it is extremely important in areas such as the Czech Republic, where current instrumentation and kits for assessing HCV infection are too costly for widespread use. We describe a new and relatively inexpensive method called nested restriction site-specific PCR (RSS-PCR) that generates a "fingerprint" pattern to represent an HCV genotype without the use of restriction endonucleases and that specifically differentiates HCV genotype 1b from the other HCV genotypes. The RSS-PCR method was applied directly to serum samples from patients with hepatitis C from the Czech Republic and from patients with known HCV genotypes from the United States. The method was validated by comparison of the subtype determined by RSS-PCR to the subtype determined from analysis of the 5' noncoding region (NC) or the nonstructural protein gene (NS5b) nucleotide sequence of HCV in these clinical samples. From 75 Czech samples containing HCV RNA, three distinct RSS-PCR patterns were observed; 54 were predicted to contain subtype 1b, 19 were predicted to contain subtype 1a, and 2 were predicted to ...Continue Reading

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Citations

Jul 20, 2002·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·D PillayUNKNOWN Public Health Laboratory Service Advisory Committee on Virology
Mar 25, 2010·Virology Journal·Sayyed H Zarkesh-EsfahaniMasoud Edalati
Mar 23, 2005·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Laura KrekulovaLee W Riley

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