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Virologica Sinica, 16 (2) : 190-192, 2001
Brief Reports
Expression of Hepatitis C Virus NS5A Gene In E .coli and Its Application in HCV Antibody Detection
Institute of Virology.Wuhan University,Hube1.Wuhan 430072.china
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
  Published online: 5 Jun 2001
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