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New combination test for hepatitis C virus genotype and viral load determination using Amplicor GT HCV MONITOR test v2.0

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World Journal of Gastroenterology 2005年第4期11卷 469-475页

作者： Motokazu Mukaide Yasuhito Tanaka Hirokazu Kakuda Kei Fujiwara Fuat Kurbanov Eturo Orito Kentaro Yoshioka Kiyotaka Fujise Shoji Harada takazumi kozaki Kazuo Takemura Kazumasa Hikiji Masashi Mizokami SRL Inc. 5-6-50ShinmachiHino-shiTokyo 191-0002Japan Department of Clinical Molecular Informative Medicine Nagoya City University Graduate School of Medical Sciences KawasumiMizuhoNagoya 467-8601Japan Department of Internal Medicine and Molecular Science Nagoya City University Graduate School of Medical Sciences KawasumiMizuhoNagoya 467-8601Japan Department of Clinical Molecular Informative Medicine Nagoya City University Graduate School of Medical Sciences KawasumiMizuhoNagoya 467-8601 Japan Third Department of Internal Medicine Nagoya University School of Medicine65 TsurumaShowaNagoya 466-8550Japan Division of Gastroenterology and Hepatology Department of Internal Medicine (Kashiwa) The Jikei University School of Medicine163-1KashiwashitaKashiwaChiba 277-8567Japan SRL Inc. 5-6-50ShinmachiHmo-shiTokyo 191-0002Japan

AIM: To develop a new sensitive and inexpensive hepatitis C virus (HCV) combination test (HCV Guideline test) that enables the determination of HCV genotypes 1, 2 and 3, and simultaneous determination of HCV viral load using commercial Amplicor GT HOV MONITOR test v2.0 (microwell version). METHODS: The HCV Guideline test used the PCR product generated in commercial Amplicor GT HCV Monitor test v2.0 for viral load measurement using microwell plate version of Amplicor HCV Monitor and also captured on separate plates containing capture probes and competitive oligonucleotide probes specific for HCV genotypes 1, 2 and 3, The HCV genotype was subsequently determined using the biotin-labeled PCR product and five biotin-labeled HCV-specific probes. RESULTS: The sensitivity of the HCV Guideline test was 0.5 KIU/mL. Specificity of the HCV Guideline test was confirmed by direct sequencing of HCV core region and molecular evolutionary analyses based on a panel of 31 samples. The comparison of the HCV Guideline test and an in-house HCV core genotyping assay using 252 samples from chronic hepatitis C patients indicated concordant results for 97.2% of samples (59.5% genotype 1, 33.7% genotype 2, 6.0% genotype 3, and 0.8% mixed genotypes). Similarly, the HCV Guideline test showed concordance with a serological test, and the serological test failed to assign any serotype in 12.7% of the samples, indicating a better sensitivity of the HCV Guideline test. CONCLUSION: Clinically, both viral load and genotypes (1, 2 and 3) have been found to be major predictors of antiviral therapy outcome regarding chronic hepatitis C based on guidelines and they are, in normal circumstances, performed as separate stand-alone assays. The HCV Guideline test is a useful method for screening large cohorts in a routine clinical setting for determining the treatment regimen and for predicting the outcome of antiviral therapy of chronic hepatitis C.

关键词： Hepatitis CVirus HCV Guideline test Viral Load Genotype

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