Antigenicity of hepatitis C virus F protein and serum prevalence of anti-F in HCV-infected patients / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 890-893, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-260560
ABSTRACT
<p><b>OBJECTIVE</b>To examine the antigenicity of hepatitis C virus (HCV) F protein and investigate serum prevalence of anti-F in HCV-infected patients.</p><p><b>METHODS</b>Eleven pairs of overlapping primers were used to synthesize the full-length HCV f gene, from which the truncated HCV f65 gene fragment was amplified by PCR. HCV f65 gene was then cloned into pET32a(+), and transformed into E. coli strain Plyss (DE3). This recombinant E.coli was induced by IPTG for the production of HCV F65 protein. The expressed HCV F65 protein, purified by Ni-NTA agarose, was further used in ELISA to detect serum anti-F, and to immunize rabbits for making polyclonal anti-F. The rabbit polyclonal anti-F was purified by Staphylococcus aureus protein A agarose.</p><p><b>RESULTS</b>After recombinant pET32a(+)-f65 was constructed successfully, HCV F65 protein was expressed and purified. The purified HCV F65 protein was used as a capture antigen in ELISA to detect serum anti-F in HCV infected patients (n = 30). The result showed that the mean A450 value and the positive rate of serum anti-F were 0.125+/-0.061 and 63.3%, respectively. The rabbit-derived polyclonal anti-F reacted specifically with HCV F65 protein, of which the titer was 130,000.</p><p><b>CONCLUSION</b>Our expressed HCV F65 protein is of antigenicity, and can be used to determine serum anti-F. Anti-F IgG does exist in the sera of the HCV-infected patients. Moreover, the rabbit-derived polyclonal anti-F can be used to detect HCV F protein.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Blood
/
Viral Envelope Proteins
/
Viral Core Proteins
/
Epidemiology
/
Prevalence
/
Hepatitis C
/
Hepacivirus
/
Hepatitis C Antigens
/
Allergy and Immunology
/
Genetics
Type of study:
Prevalence study
/
Risk factors
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Hepatology
Year:
2006
Type:
Article
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