Quantitation of plasma human immunodeficiency virus type 1 RNA by competitive polymerase chain reaction.
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-1 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-1 gag region sequence. The mutant cRNA (K4) was shown to amplify with efficiency equivalent to that of wild-type HIV-1. Known quantities of K4 cRNA added to wild-type HIV-1 in a competitive PCR strategy using a radiolabeled primer permitted quantitation of wild-type HIV-1 RNA over four orders of magnitude (10(3)-10(6) RNA copies). RNA isolated from plasma from AIDS patients yielded 10(3) to 8 x 10(4) HIV-1 RNA copies/ml of plasma with an average intrasample coefficient of variation of .26. This method offers a sensitive assay with a broad dynamic range for monitoring HIV-1 activity in the plasma of AIDS patients. It may provide a useful tool for assessing the effects of antiretroviral therapy.
Scadden
D T
DT
Division of Hematology/Oncology, New England Deaconess Hospital, Harvard Medical School, Boston, Massachusetts.
Wang
Z
Z
Groopman
J E
JE
D001667:Binding, Competitive; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D012367:RNA, Viral