[Overexpression, purification of recombinant HIV-1 gp41 protein and detection of HIV antibody in urine].
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%.


CONCLUSIONS
The HIV-1 urine antibody kit can be used in screening and diagnosing for HIV-1 infection.
Zhang
Xiao-Guang
XG
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Qi
Qi-Ping
QP
Ma
Jing
J
Zhang
Xiao-Mei
XM
Wang
Zi-Chun
ZC
Li
Hong-Xia
HX
Zeng
Yi
Y
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015483:HIV Antibodies / Q000652:urine*; D016655:HIV Core Protein p24 / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D015658:HIV Infections / Q000276:immunology* / Q000652:urine; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D015483:HIV Antibodies; D016655:HIV Core Protein p24; D011994:Recombinant Proteins