Genotyping of hepatitis C virus by the polymerase chain reaction: a comparison of two PCR-based methods using core or NS5 primers

Hashimoto, Michie; Chayama, Kazuaki; Tubota, Akihito; Koida, Isao; Saitoh, Satosi; Arase, Yasuji; Ikeda, Kenji; Kobayashi, Mariko; Kanda, Masumi; Iwasaki, Satomi; Nakano, Akemi; Takagi, Kimiko; Koike, Hiromi; Kumada, Hiromitsu

doi:10.1016/0928-4346(95)00216-6

Cited by 3 publications

(1 citation statement)

References 4 publications

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“…3,4 Quantification was performed with previously determined HCV RNA-positive serum as a standard and the results were highly reproducible. We determined the HCV genotype with NS 5 region PCR using the method described by Chayama et al, 11 and Hashimoto et al 12 Statistical analysis was done with the Wilcoxon signed rank sum test and Fisher's exact test and results were considered significant when P Ͻ 0.05 by two-tailed test.…”

Section: Methodsmentioning

confidence: 99%

Usefulness of hepatitis C virus RNA counts by second generation HCV bDNA-probe in chronic hepatitis C based on the HCV genotype

Kobayashi

Kumada

Arase

et al. 1998

Journal of Gastroenterology

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Detection of hepatitis C virus (HCV) RNA by a second generation (ver 2) HCV bDNA-probe method (bDNA-probe) was compared with detection by the first generation (ver 1) assay. The two assays were performed simultaneously with the same serum samples of HCV genotypes 1b, 2a, 2b, 3a, and 3b. The positive rates with ver 1 were 82% for HCV genotype 1b (type 1b), 57.6% for HCV genotype 2a (type 2a), 75.0% for HCV genotype 2b (type 2b), 55.6% for HCV genotype 3a (type 3a), and 93.8% for HCV genotype 3b (type 3b). The positive rates with ver 2 were 95.0% for type 1b, 93.9% for type 2a, 83.3% for type 2b, 100% for type 3a, and 93.8% for type 3b. With Fisher's exact test, the detection rate for type 2a was significantly higher (P = 0.001) with ver 2 than with ver 1. We obtained regression lines using the HCV counts measured by bDNA-probe on the y axis and the HCV counts obtained by an HCV reverse transcriptase (RT)-competitive polymerase chain reaction method (competitive PCR) on the x axis. The gradients for types 1b, 2a, and 3b were greater with ver 2 compared to ver 1. The gradients for types 2a and 3b were the highest: for type 2a, y = 0.135x + 0.6 with ver 1 and y = 0.248x + 0.1 with ver 2; for type 3b, y = 0.366x + 0.1 with ver 1 and y = 0.727x + 0.3 for ver 2. In addition, HCV-RNA counts for all the genotypes tested in this study were significantly higher with ver 2 than with ver 1. Hence, we conclude that ver 2 of the bDNA-probe measures HCV-RNA counts closer to those obtained with competitive PCR than the ver 1 assay.

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Section: Methodsmentioning

confidence: 99%

Usefulness of hepatitis C virus RNA counts by second generation HCV bDNA-probe in chronic hepatitis C based on the HCV genotype

Kobayashi

Kumada

Arase

et al. 1998

Journal of Gastroenterology

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Fluctuation patterns of HCV-RNA serum level in patients with chronic hepatitis C

Arase

Ikeda

Chayama

et al. 2000

Journal of Gastroenterology

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The serum level of hepatitis C virus (HCV)-RNA is clinically important as a predictor of the response to interferon (IFN) therapy in patients with chronic hepatitis C. If serum HCV-RNA levels fluctuate during follow-up, and IFN therapy is begun at the time of a low HCV-RNA level, the IFN therapy may be more effective. We evaluated the fluctuation of HCV-RNA serum levels for 2 years in 212 patients with chronic hepatitis C, untreated with IFN who had HCV genotype 1b and an HCV-RNA level of 10 Meq/ml or more at first consultation. The HCV-RNA level was measured monthly for 2 years with an HCV branched DNA probe assay (b DNA probe assay). We classified HCV-RNA patterns into three types by the ratio of maximum HCV-RNA level (a) to minimum HCV-RNA level (b). In pattern 1 (constant type, 151 patients; 71.2%) the a/ b ratio was 1-5. In pattern 2 (slight fluctuation type, 46 patients; 21.7%) the a/b ratio was 5-10. In pattern 3 (severe fluctuation type, 15 patients; 7.1%), the a/b ratio was 10 or more. Next, we evaluated the factors associated with the three patterns. Acute exacerbation of chronic hepatitis was regarded as an increase in serum alanine aminotransferase (ALT) level to more than 250 IU/l. The incidence of acute exacerbation for a 2-year follow-up was 13.9% (21/151) in pattern 1, 19.6% (9/46) in pattern 2, and 53.3% (8/15) in pattern 3. Multivariate analysis showed that acute exacerbation was the most important factor in the manifestation pattern 3. In conclusion, we found that: (1) about 70% of patients had a constant HCV-RNA levels for 2 years. (2) A few patients had severe fluctuation of serum HCV-RNA level after acute exacerbation of chronic hepatitis.

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Typing six major hepatitis C virus genotypes by polymerase chain reaction using primers derived from nucleotide sequences of the NS5 region

Hashimoto

Chayama

Tubota

et al. 1996

International Hepatology Communications

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Genotyping of hepatitis C virus by the polymerase chain reaction: a comparison of two PCR-based methods using core or NS5 primers

Cited by 3 publications

References 4 publications

Usefulness of hepatitis C virus RNA counts by second generation HCV bDNA-probe in chronic hepatitis C based on the HCV genotype

Usefulness of hepatitis C virus RNA counts by second generation HCV bDNA-probe in chronic hepatitis C based on the HCV genotype

Fluctuation patterns of HCV-RNA serum level in patients with chronic hepatitis C

Typing six major hepatitis C virus genotypes by polymerase chain reaction using primers derived from nucleotide sequences of the NS5 region

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