QUANTIFICATION OF HIV-1 USING MULTIPLE COMPETITORS IN A SINGLE-TUBE ASSAY

Methods for quantification of human immunodeficiency virus type I (HN- I) based on competitive PCR and fragment analysis have been developed. Samples containing HN-I DNA and known amounts of three cloned competi tors were co-amplified by PCR with semi-nested primers. The competitor DNAs contained the same long terminal repeat primer binding sequences as the wild-type DNA, but they are different in internal sequences an d length. One of the inner primers was fluorescent-labeled to allow di scrimination between the wild-type DNA and the three competitors by fr agment analysis using a standard automated sequencer: A calibration cu rve using the peak area of the three competitors enabled accurate dete rmination of target amount with minimal variations. The method present ed here can be used for quantification of HIV-1 in clinical samples an d will be useful for monitoring disease progression and treatment effe cts.