Establishment of a quantification method for the amino acid 70 substitution in the core region of hepatitis C virus genotype 1b

VernacularTitle:采用TaqMan MGB探针定量检测HCV 1b基因型C区氨基酸70变异
Author: Zhongjie HU ¹ ; Ying LIU ; Lixia QIU ; Zuopeng FAN ; Wei NIE ; Shan LIANG
Author Information

1. 100069,首都医科大学附属北京佑安医院丙肝与中毒性肝病科
Keywords: Hepatitis viruses; Reverse transcription polymerase chain reaction; DNA probes,MGB
From: Chinese Journal of Experimental and Clinical Virology 2014;28(4):305-308
CountryChina
Language:Chinese
Abstract: Objective To develop a rapid and sensitive method to quantify the substitution of amino acid (aa) 70 in hepatitis C virus (HCV) genotype 1 b core region.Methods Specific primers and TaqManminor groove binder probes were designed to establish a real time reverse transcription-polymerase chain reaction for aa70 substitution.The specificity,sensitivity and accuracy of the system were tested.Results The probes could clearly distinguish 70W and 70M.The detection limit for each system was 5 copies/ reaction.Further tests using cloning sequencing confirmed the reliability of the detection system.Conclusions A novel system was developed and provides an effective,sensitive,and accurate approach to quantify the wild type and mutant strains of HCV 1b core region as70.