BACKGROUND: Recent studies have focused on whether different hepatitis C virus (HCV) genotypes are associated with different profiles of pathogenicity, infectivity, and response to antiviral therapy. We needed to develop a convenient screening test for HCV genotypes 1 and 2. METHOD: We tested 55 patients with known chronic HCV infection. Viral RNA was extracted from serum samples using an automatic viral RNA purification system, and HCV genotypes were determined by reverse transcriptase-polymerase chain reaction using LightCycler melting curve analysis with SYBR Green I. RESULTS: HCV RNA was detected in all samples and each genotype was determined. The mean (standard deviation) melting temperatures for subtypes 1b (n = 32), 2a (n = 15) and 2b (n = 8) were 93.14 degrees C (0.51 degrees C), 91.08 degrees C (0.49 degrees C) and 91.77 degrees C (0.27 degrees C), respectively. Genotypes 1 and 2 were differentiated within 3 h by this method. CONCLUSIONS: Our melting curve analysis is a rapid and convenient screening test for differential identification of HCV genotypes 1 and 2.
BACKGROUND: Recent studies have focused on whether different hepatitis C virus (HCV) genotypes are associated with different profiles of pathogenicity, infectivity, and response to antiviral therapy. We needed to develop a convenient screening test for HCV genotypes 1 and 2. METHOD: We tested 55 patients with known chronic HCV infection. Viral RNA was extracted from serum samples using an automatic viral RNA purification system, and HCV genotypes were determined by reverse transcriptase-polymerase chain reaction using LightCycler melting curve analysis with SYBR Green I. RESULTS:HCV RNA was detected in all samples and each genotype was determined. The mean (standard deviation) melting temperatures for subtypes 1b (n = 32), 2a (n = 15) and 2b (n = 8) were 93.14 degrees C (0.51 degrees C), 91.08 degrees C (0.49 degrees C) and 91.77 degrees C (0.27 degrees C), respectively. Genotypes 1 and 2 were differentiated within 3 h by this method. CONCLUSIONS: Our melting curve analysis is a rapid and convenient screening test for differential identification of HCV genotypes 1 and 2.
Authors: Linda Cook; KaWing Sullivan; Elizabeth M Krantz; Arthur Bagabag; Keith R Jerome Journal: J Clin Microbiol Date: 2006-09-20 Impact factor: 5.948
Authors: Sueli M Nakatani; Carlos A Santos; Irina N Riediger; Marco A Krieger; Cesar A B Duarte; Marco A Lacerda; Alexander W Biondo; Flair J Carrilho; Flair J Carilho; Suzane K Ono-Nita Journal: PLoS One Date: 2010-04-13 Impact factor: 3.240