Citation: RUAN Hua, GAO Jin—rang, YE Lin hai, XU Jing—ping, W ANG Xiao-ling.ZHAO Yue-e.W U Zheng—hui, . Expression of Hepatitis C Virus NS5A Gene In E .coli and Its Application in HCV Antibody Detection .VIROLOGICA SINICA, 2001, 16(2) : 190-192.

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

  • Available online: 05 June 2001
  • Full-length NS5A gene of the hepatitis C virus was amplified by PCR using plasmid pBAC25 containing HCV nonstructural gene as template.The am plified fragment(about 1.34 kb)was cloned into plaS d pQE32.and the recombinant plasmid pQENSSA was expressed in JM109 strain The NS5A protein WaS purified by NiS04 metal chelating resin,and characterized by W estern—blot Its antigenecity was de~ermined 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 stan— dand sera,and the negative rate of anti—NS5A was 10O% (40/40)in fourty negative standand sera The results showed that the Full—length NSSA 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.

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

    • 1. Institute of Virology.Wuhan University,Hube1.Wuhan 430072.china

    Abstract: Full-length NS5A gene of the hepatitis C virus was amplified by PCR using plasmid pBAC25 containing HCV nonstructural gene as template.The am plified fragment(about 1.34 kb)was cloned into plaS d pQE32.and the recombinant plasmid pQENSSA was expressed in JM109 strain The NS5A protein WaS purified by NiS04 metal chelating resin,and characterized by W estern—blot Its antigenecity was de~ermined 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 stan— dand sera,and the negative rate of anti—NS5A was 10O% (40/40)in fourty negative standand sera The results showed that the Full—length NSSA 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.

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