DETECTION OF REVERSE-TRANSCRIPTASE BY A HIGHLY SENSITIVE ASSAY IN SERA FROM PERSONS INFECTED WITH HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1

In an ultrasensitive assay for reverse transcriptase (RT), an in vitro -transcribed heteropolymeric RNA sequence was used as a template and p olymerase chain reaction (PCR) amplification with Southern blot hybrid ization served as a detection system for the cDNA reaction product, Th e assay, called Amp-RT, detected 9 tested retroviruses in unconcentrat ed culture supernatants diluted 10(2)- to 10(5)-fold. A comparative an alysis using human immunodeficiency virus type 1 (HIV-1) revealed that Amp-RT was 100,000 times more sensitive than the standard RT assay, 1 0,000 times more sensitive than p24 antigen capture and branched DNA a ssays, and 100 times more sensitive than RT-PCR or TCID50 assays, Anal ysis of serum specimens from 42 HIV-1-infected persons by Amp-RT showe d that 36 samples (85.7%) were RT-positive. In contrast, 41 serum spec imens from persons seronegative for HIV-I and human T lymphotropic vir us types I and II were all Amp-RT-negative.