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 polymerase chain reaction (PCR) amplification with Southern blot hybridization served as a detection system for the cDNA reaction product. The assay, called Amp-RT, detected 9 tested retroviruses in unconcentrated culture supernatants diluted 10(2)- to 10(5)-fold. A comparative analysis using human immunodeficiency virus type 1 (HIV-1) revealed that Amp-RT was 100,000 times more sensitive than the standard RT assay, 10,000 times more sensitive than p24 antigen capture and branched DNA assays, and 100 times more sensitive than RT-PCR or TCID50 assays. Analysis of serum specimens from 42 HIV-1-infected persons by Amp-RT showed that 36 samples (85.7%) were RT-positive. In contrast, 41 serum specimens from persons seronegative for HIV-1 and human T lymphotropic virus types I and II were all Amp-RT-negative.
Heneine
W
W
Retrovirus Diseases Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
Yamamoto
S
S
Switzer
W M
WM
Spira
T J
TJ
Folks
T M
TM
D000818:Animals; D001483:Base Sequence; D018791:CD4 Lymphocyte Count; D018076:DNA, Complementary / Q000096:biosynthesis; D004279:DNA, Viral / Q000097:blood; D004686:Encephalomyocarditis virus / Q000235:genetics; D016655:HIV Core Protein p24 / Q000097:blood; D015658:HIV Infections / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000097:blood*; D012190:Retroviridae / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013698:Templates, Genetic
D018076:DNA, Complementary; D004279:DNA, Viral; D016655:HIV Core Protein p24; D012367:RNA, Viral; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase