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Clinical Chemistry 39: 244-247, 1993;
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Clinical Chemistry, Vol 39, 244-247, Copyright © 1993 by American Association for Clinical Chemistry

Nonradioactive, colorimetric microplate hybridization assay for detecting amplified human immunodeficiency virus DNA

JM Rapier, Y Villamarzo, G Schochetman, CY Ou, CL Brakel, J Donegan, W Maltzman, S Lee, D Kirtikar and D Gatica
Division of HIV/AIDS, Centers for Disease Control, Atlanta, GA 30333.

A nonradioactive, colorimetric microplate hybridization procedure was used to assay human immunodeficiency virus (HIV) DNA, amplified by the polymerase chain reaction (PCR). Under the PCR conditions used, four proviral copies per 150,000 cells were detected by amplifying a series of DNA mixtures that contained various copy numbers of HIV. Assays of PCR-amplified DNA from peripheral blood mononuclear cells of seronegative individuals yielded negative results (104 of 104), whereas samples from seropositive individuals yielded > 99% positive results (141 of 142). Similar results were obtained in a chemiluminescent assay with an acridinium ester-labeled probe and in a solution hybridization assay in which a 32P-labeled probe was used.




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Copyright © 1993 by the American Association for Clinical Chemistry.