Detection of polymerase chain Reaction-Amplified Immunodeficiency virus proviral DNA with a digoxigenin-labeled nucleotide and Enzyme-Linked oligosorbent assay

An enzyme-linked oligosorbent assay (ELOSA) was developed for the detection on microtiter plates of PCR amplified human immunodeficiency DNA. In this nonisotopic PCR assay designated PCR-ELOSA, fragment of the HIV gag gene from peripheral blood mononuclear cells (PBMCs) was first amplified with a digoxigenin-labeled amplified DNA segment was reacted in solution with a probe labeled with biotin. Hybrids were captured in microtiter plate coated with streptavidin. Specific bound hybrids were detected by the addition of on enzyme-labeled antibody against digoxigenin and chromogenic substrate. This method was developed to increase both sensitivity of detection of PCR product.

کلیدواژه ها:

HIV ، Nested PCR ، PCR- ELOSA