title: Genotyping of hepatitis C virus by melting curve analysis with SYBR Green I.
abstract: 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.
  
  
  METHODS
  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.affiliation: Department of Clinical Laboratory Medicine, Gifu University School of Medicine, Gifu 500-8705, Japan. g2104021@gueda.cc.gifu-u.ac.jp
authors.forename: Hidetsugu
authors.initials: H
authors.lastname: Fujigaki
authors.forename: Masao
authors.initials: M
authors.lastname: Takemura
authors.forename: Kanako
authors.initials: K
authors.lastname: Takahashi
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yamada
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Fujii
authors.forename: Hisayasu
authors.initials: H
authors.lastname: Wada
authors.forename: Kuniaki
authors.initials: K
authors.lastname: Saito
authors.forename: Hiroo
authors.initials: H
authors.lastname: Ohnishi
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Seishima
mesh_terms: D052160:Benzothiazoles; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics*; D003959:Diamines; D016679:Genome, Viral; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000472:pathogenicity; D006526:Hepatitis C / Q000097:blood / Q000235:genetics*; D006801:Humans; D009691:Nucleic Acid Denaturation / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000235:genetics / Q000379:methods; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral / Q000097:blood / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014774:Virulence / Q000235:genetics
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D018076:DNA, Complementary; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; C098022:SYBR Green I