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M9480100.TXT
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1994-08-09
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Document 0100
DOCN M9480100
TI Identification of low-level contamination of blood as basis for
detection of human immunodeficiency virus (HIV) DNA in anti-HIV-negative
specimens.
DT 9410
AU Sabino EC; Delwart E; Lee TH; Mayer A; Mullins JI; Busch MP; Irwin
Memorial Blood Centers, San Francisco, CA 94118.
SO J Acquir Immune Defic Syndr. 1994 Aug;7(8):853-9. Unique Identifier :
AIDSLINE GENBANK/L20380
AB The significance of detection of human immunodeficiency virus (HIV) DNA
by the polymerase chain reaction (PCR) in seronegative or seroconverting
(SC) subjects remains controversial. In a previously reported study, we
identified a case in which a specimen collected 12 months before
seroconversion (pre-SC) was found repeatedly to be PCR positive in three
experienced laboratories, while the 6-month pre-SC bleed was
PCR-negative; PCR-based human leukocyte antigen (HLA)-DQA and -DRB
typing of serial peripheral blood mononuclear cell (PBMC) samples from
this case did not indicate a specimen mix-up or labeling error. To
further investigate this case, we used HIV env sequence and DNA
heteroduplex gel-shift analyses to characterize HIV quasispecies present
in serial pre- and post-SC specimens. HIV env sequences and gel-shift
pattern analyses from the 12-month pre-SC versus post-SC samples
indicated that markedly distinct quasispecies were present, suggesting
possible abortive infection followed by reinfection and subsequent
seroconversion. However, the HIV burden of this pre-SC sample was very
low (1 provirus/10(6) PBMCs), and the quasispecies was highly
heterogeneous, findings suggesting long-term rather than recent HIV
infection. To test the hypothesis that the index pre-SC sample was PCR
positive owing to trace blood contamination during initial processing,
we analyzed the three seropositive samples collected on the same date in
1985. One of these samples was highly related to the index pre-SC sample
by env sequence and gel-shift methodologies.(ABSTRACT TRUNCATED AT 250
WORDS)
DE Base Sequence Blotting, Western Cell Separation Consensus Sequence
Diagnostic Errors DNA Primers/CHEMISTRY DNA, Viral/*BLOOD/CHEMISTRY
Gene Products, env/GENETICS/IMMUNOLOGY Human HIV Antibodies/*BLOOD
HIV Envelope Protein gp120/GENETICS *HIV Seronegativity HIV
Seropositivity/*DIAGNOSIS HIV-1/*GENETICS/IMMUNOLOGY/ISOLATION & PURIF
HLA-DR2 Antigen/GENETICS Leukocytes, Mononuclear/MICROBIOLOGY
Molecular Sequence Data Peptide Fragments/GENETICS Polymerase Chain
Reaction Protein Precursors/IMMUNOLOGY Sequence Homology, Nucleic Acid
Support, U.S. Gov't, P.H.S. JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).