title: Absence of HIV-1 DNA in high-risk seronegative individuals using high-input polymerase chain reaction.
abstract: Evidence of frequent HIV-1 infections in antibody-negative, high-risk individuals (so-called 'silent' infections) remains controversial. To evaluate whether these discrepant results may be the consequence of intermittent detection of rare infected cells (low viral load) preceding seroconversion, we developed a modification of the polymerase chain reaction (PCR) technique which enabled analysis of 10-fold greater amounts of cellular DNA per reaction than standard PCR (2 x 10(6) rather than 0.2 x 10(6) input cells). This technique allowed consistent detection of HIV-1 provirus in two seropositive individuals who had repeatedly tested negative by standard-input PCR. However, results were negative when high-input PCR was applied to 51 specimens from 39 selected high-risk seronegative individuals. These results suggest that variations in viral load preceding or in the absence of seroconversion probably do not explain discrepant evidence regarding silent HIV-1 infection.
authors.affiliation: Irwin Memorial Blood Centers, San Francisco, California 94118.
authors.forename: T H
authors.initials: TH
authors.lastname: Lee
authors.forename: Z
authors.initials: Z
authors.lastname: el-Amad
authors.forename: M
authors.initials: M
authors.lastname: Reis
authors.forename: M
authors.initials: M
authors.lastname: Adams
authors.forename: E A
authors.initials: EA
authors.lastname: Donegan
authors.forename: T R
authors.initials: TR
authors.lastname: O'Brien
authors.forename: A R
authors.initials: AR
authors.lastname: Moss
authors.forename: M P
authors.initials: MP
authors.lastname: Busch
mesh_terms: D015331:Cohort Studies; D004279:DNA, Viral / Q000302:isolation & purification*; D015658:HIV Infections / Q000209:etiology / Q000382:microbiology*; D006679:HIV Seropositivity / Q000382:microbiology*; D015497:HIV-1 / Q000472:pathogenicity*; D006716:Homosexuality; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D012189:Retrospective Studies; D012307:Risk Factors; D012725:Sexual Behavior
chemical_list: D004279:DNA, Viral