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J Clin Microbiol. 1989 November; 27(11): 2570-2573

Detection of enzymatically amplified human immunodeficiency virus DNA by oligonucleotide solution hybridization and by incorporation of radiolabeled deoxynucleotides.

Medical Research Council AIDS Virus Research Unit, University of the Witwatersrand, Johannesburg, South Africa.

ABSTRACT

Two methods are described for the detection of in vitro enzymatically amplified DNA. The first method involves solution hybridization of labeled oligonucleotides to amplified products. Hybridization of primers to a dilution series of known concentration of amplified DNA showed that approximately 5 pg of DNA could be detected by this method. In the second method, radiolabeled deoxynucleotides were incorporated into the elongating DNA chain. Both methods were able to detect amplified products 10 cycles before detection by ethidium bromide staining. Some variations of these techniques are discussed.