Detection of human recombinant HIV-p24 antigen by Immuno-polymerase chain reaction
- VernacularTitle:免疫PCR检测HIV-p24抗原的实验研究
- Author:
Ji ZHENG
;
Weiling FU
;
Tianlun JIANG
- Publication Type:Journal Article
- Keywords:
Immuno-PCR;
Sensitivity;
Human recombinant HIV-p24 antigen;
AIDS;
Blood donor;
Early screening
- From:
Chinese Journal of Blood Transfusion
1988;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To describe a highly sensitive immuno-polymerase chain reaction (immuno-PCR) assay for the detection of human recombinant HIV-p24 antigen. Methods We used gold-magnetic particles as the carriers,mouse anti-p24 monoclonal antibody as the capture antibody and biotinylated goat anti-p24 polyclonal antibody as the detection antibody. The reporter DNA was initially generated by PCR amplification using a biotinylated primer,and was bound with streptavidin to biotinylated polyclonal antibody. Human recombinant p24 antigen sandwiched by antibodies was detected by amplifying the reporter DNA using PCR. The optimal concentration of sreptavidin and DNA label were determined using square titration. The electrophoresis gels were imaged and analyzed by Quantity One software. Results The optimal concentration of sreptavidin and DNA label were determined to be 0.1 mg/ L and 10 ng/ L,respectively. The detection limit of the immuno-PCR assay was 0.1 ng/L,higher than that of the conventional ELISA. Conclusion A highly sensitive immuno-PCR for human recombinant HIV-p24 antigen was indicated to be an available method for early screening of HIV infectors in blood donors.
