Overexpression, Purification of recombinant HIV-1 gp41 protein and detection of HIV antibody in urine
10.3760/cma.j.issn.1003-9279.2008.04.023
- VernacularTitle:高效表达重组HIV-1 gp41及在尿液检测中的应用
- Author:
Xiao-Guang ZHANG
1
;
Qi-Ping QI
;
Jing MA
;
Xiao-Mei ZHANG
;
Zi-Chun WANG
;
Hong-Xia LI
;
Yi ZENG
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
HIV;
gp41 overexpression;
purification;
Antibdies,viral;
Urine
- From:
Chinese Journal of Experimental and Clinical Virology
2008;22(4):308-310
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a specific and sensitive Enzyme-linked immunosorbent Assay(ELISA)kit for detection of HIV-1 antibody in urine using Escherichia coli expression products as coating antigen. Methods The truncated HIV-1 gp41 gene fragment of major antigenic epitopes was inserted into the plasmid pET22b to obtain expression plasmid pET22b-mgp41. The recombinant antigen was expressed in BL21 (DE3) strains of Escherichia coli and was purified by immobilized metal chelation and gel filtration chromatography. Using this antigen as coating antigen, a HIV-1 urine antibody ELISA kit was developed. In order to examine the clinical utility of the kit, 5437 urine samples were assayed, which consisted of 641 urine samples from HIV infected patients and 4796 samples from normal subjects. Results The purity of purified antigen is up to 95%. Anti-HIV antibodies were detected in all the urine samples from HIV infected patients, and the diagnostic sensitivity for HIV-1 infection was 100%. In healthy control group, 71 cases showed false positive, the specificity was 98.52%.Conclusion The HIV-1 urine antibody kit can be used in screening and diagnosing for HIV-1 infection.
