Identification of low-level contamination of blood as basis for detection of human immunodeficiency virus (HIV) DNA in anti-HIV-negative specimens.
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)
Irwin Memorial Blood Centers, San Francisco, CA 94118.
E C
EC
Sabino
E
E
Delwart
T H
TH
Lee
A
A
Mayer
J I
JI
Mullins
M P
MP
Busch
D001483:Base Sequence; D015153:Blotting, Western; D002469:Cell Separation; D016384:Consensus Sequence; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000097:blood* / Q000737:chemistry; D003951:Diagnostic Errors; D015686:Gene Products, env / Q000235:genetics / Q000276:immunology; D015483:HIV Antibodies / Q000097:blood*; D015699:HIV Envelope Protein gp120 / Q000235:genetics; D019302:HIV Envelope Protein gp160; D018023:HIV Seronegativity; D006679:HIV Seropositivity / Q000175:diagnosis*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D015802:HLA-DR2 Antigen / Q000235:genetics; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D004279:DNA, Viral; D015686:Gene Products, env; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; C059762:HIV envelope protein gp120 (305-321); D015802:HLA-DR2 Antigen; D010446:Peptide Fragments; D011498:Protein Precursors