TY - JOUR
T1 - Genotyping of hepatitis C virus by melting curve analysis with SYBR Green I
AU - Fujigaki, Hidetsugu
AU - Takemura, Masao
AU - Takahashi, Kanako
AU - Yamada, Yasuhiro
AU - Fujii, Hidehiko
AU - Wada, Hisayasu
AU - Saito, Kuniaki
AU - Ohnishi, Hiroo
AU - Seishima, Mitsuru
PY - 2004/3
Y1 - 2004/3
N2 - 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 lb (n = 32), 2a (n = 15) and 2b (n = 8) were 93.14°C (0.51°C), 91.08°C (0.49°C) and 91.77°C (0.27°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.
AB - 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 lb (n = 32), 2a (n = 15) and 2b (n = 8) were 93.14°C (0.51°C), 91.08°C (0.49°C) and 91.77°C (0.27°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.
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U2 - 10.1258/000456304322880014
DO - 10.1258/000456304322880014
M3 - Article
C2 - 15025803
AN - SCOPUS:1942438005
SN - 0004-5632
VL - 41
SP - 130
EP - 132
JO - Annals of Clinical Biochemistry
JF - Annals of Clinical Biochemistry
IS - 2
ER -
