Title Hepatitis C Virus (HCV) Genotyping by Annealing Reverse Transcription-PCR Products with Genotype-Specific Capture Probes
Author Jungmin Rho1, Jong Soon Ryu2, Wonhee Hur3, Chang Wook Kim2,3, Jeong Won Jang2,3, Si Hyun Bae2,3, Jong Young Choi2,3, Sung Key Jang1,4, and Seung Kew Yoon2,3*
Address 1Panbionet, Corp., POSTECH Biotech Center, Pohang 790-784, Republic of Korea, 2WHO Collaborating Center on Viral Hepatitis, College of Medicine, The Catholic University of Korea, Seoul 137-040, Republic of Korea, 3Division of Hepatology, Department of Internal Medicine, The Catholic University of Korea, Seoul 137-040, Republic of Korea, 4Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea
Bibliography Journal of Microbiology, 46(1),81-87, 2008,
DOI
Key Words hepatitis C virus, genotype, reverse-hybridization
Abstract The genotype of the hepatitis C virus (HCV) strain infecting a given patient is an important predictive factor for the clinical outcome of chronic liver disease and its response to anti-viral therapeutic agents. We herein sought to develop a new easy, sensitive and accurate HCV genotyping method using annealing genotype- specific capture probes (AGSCP) in an automation-friendly 96-well plate format. The validation of our new AGSCP was performed using the Standard HCV Genotype Panel. We then used both our AGSCP and the commercially available INNO-LiPA assay to analyze the HCV genotypes from 111 Korean patients. Discordant results were analyzed by direct sequencing. AGSCP successfully genotyped the standard panel. The genotypes of 111 patient samples were also obtained successfully by AGSCP and INNO-LiPA. We observed a high concordance rate (93 matched samples, 83.8%) between the two assays. Sequencing analysis of the 18 discordant results revealed that the AGSCP had correctly identified 12 samples, whereas the INNO- LiPA had correctly identified only 6. These results collectively indicate that AGSCP assay is a convenient and sensitive method for large-scale genotyping, and it may be a promising tool for the determination of HCV and other genotypes in clinical settings.