Genotyping of hepatitis C virus by melting curve analysis with SYBR Green I

Hidetsugu Fujigaki, Masao Takemura, Kanako Takahashi, Yasuhiro Yamada, Hidehiko Fujii, Hisayasu Wada, Kuniaki Saito, Hiroo Ohnishi, Mitsuru Seishima

Research output: Contribution to journalArticle

7 Citations (Scopus)

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. 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.

Original languageEnglish
Pages (from-to)130-132
Number of pages3
JournalAnnals of Clinical Biochemistry
Volume41
Issue number2
DOIs
Publication statusPublished - 01-03-2004

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Viruses
Hepacivirus
Freezing
Melting
Genotype
Viral RNA
Screening
Polymerase chain reaction
RNA-Directed DNA Polymerase
Chronic Hepatitis C
Virus Diseases
Reverse Transcriptase Polymerase Chain Reaction
Purification
Antiviral Agents
Virulence
Melting point
SYBR Green I
RNA
Temperature
Serum

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

Cite this

Fujigaki, H., Takemura, M., Takahashi, K., Yamada, Y., Fujii, H., Wada, H., ... Seishima, M. (2004). Genotyping of hepatitis C virus by melting curve analysis with SYBR Green I. Annals of Clinical Biochemistry, 41(2), 130-132. https://doi.org/10.1258/000456304322880014
Fujigaki, Hidetsugu ; Takemura, Masao ; Takahashi, Kanako ; Yamada, Yasuhiro ; Fujii, Hidehiko ; Wada, Hisayasu ; Saito, Kuniaki ; Ohnishi, Hiroo ; Seishima, Mitsuru. / Genotyping of hepatitis C virus by melting curve analysis with SYBR Green I. In: Annals of Clinical Biochemistry. 2004 ; Vol. 41, No. 2. pp. 130-132.
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Fujigaki, H, Takemura, M, Takahashi, K, Yamada, Y, Fujii, H, Wada, H, Saito, K, Ohnishi, H & Seishima, M 2004, 'Genotyping of hepatitis C virus by melting curve analysis with SYBR Green I', Annals of Clinical Biochemistry, vol. 41, no. 2, pp. 130-132. https://doi.org/10.1258/000456304322880014

Genotyping of hepatitis C virus by melting curve analysis with SYBR Green I. / Fujigaki, Hidetsugu; Takemura, Masao; Takahashi, Kanako; Yamada, Yasuhiro; Fujii, Hidehiko; Wada, Hisayasu; Saito, Kuniaki; Ohnishi, Hiroo; Seishima, Mitsuru.

In: Annals of Clinical Biochemistry, Vol. 41, No. 2, 01.03.2004, p. 130-132.

Research output: Contribution to journalArticle

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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

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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.

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