Expression of Hepatitis C Virus NS5A Gene In E.coli and Its Application in HCV Antibody Detection

VernacularTitle:HCV NS5A基因表达及其在血清学检测中的应用评估
Author: Hua, RUAN ; Jin-rong, GAO ; Lin-Bai, YE ; Jing-ping, XU ; Xiao-ling, WANG ; Yue-e, ZHAO ; Zheng-hui, WU
Publication Type:Journal Article
From:Virologica Sinica 2001;16(2):190-192
CountryChina
Language:Chinese
Abstract: Full-length NS5A gene of the hepatitis C virus was amplified by PCR using plasmid pBAC25 containing HCV nonstructural gene as template. The amplified fragment (about 1.34 kb) was cloned into plasmid pQE32, and the recombinant plasmid pQENS5A was expressed in JM109 strain. The NS5A protein was purified by NiSO4 metal chelating resin, and characterized by Western-blot. Its antigenecity was determined by ELISA. The positive detection rate of anti-NS5A was 75% (69/92) in ninety-two clinic sera. The positive rate of anti-NS5A was 82.5% (33/40) in fourty positive standand sera, and the negative rate of anti-NS5A was 100% (40/40) in fourty negative standand sera. The results showed that the Full-length NS5A proteinn had the higher sensitivity and specificity in the detection of HCV antibody in sera, we suggested that NS5A protein was a useful antigen for blood screening.