Quantitation of Plasma Human Immunodeficiency Virus Type 1 RNA by Competitive Polymerase Chain Reaction

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Abstract
Clinical measures of human immunodeficiency virus (HIV) type 1 activity in.vivo are limited and hinder the assessment of antiretroviral therapies. Reported here is a method for quantitating HIV-I RNA in human plasma using the polymerase chain reaction (PCR). This method uses an internal cRNA standard generated from a cloned 113-bp deletion mutation of a highly conserved HIV-I gag region sequence. The mutant cRNA (K4)was shown to amplify with efficiency equivalent to that of wild-type HIV-l. Known quantities ofK4 cRNA added to wild-type HIV-I in a competetive PCR strategy using a radiolabeled primer permitted quantitation of wild-type HIV1 RNA over four orders of magnitude (103-106 RNA copies). RNA isolated from plasma from AIDS patients yielded 103 to 8 × 104 HIV-I RNA copies/ml of plasma with an average intrasampie coefficient of variation of .26. This method offers a sensitive assay with a broad dynamic range for monitoring HIV-I activity in the plasma of AIDS patients. It may provide a useful tool for assessing the effects of antiretroviral therapy.

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