Inhibition of human immunodeficiency virus gene amplification by heparin.
Gene amplification of virus-specific sequences is widely used as a method to detect or confirm human immunodeficiency virus (HIV) infection. In this study we used an enzyme-linked affinity assay to quantify polymerase chain reaction products from whole blood, plasma, and separated mononuclear cells collected in the presence of four common anticoagulants: acid citrate dextrose, sodium EDTA, potassium oxalate, and sodium heparin. Attenuation of the product signal was observed after amplification of nucleic acid extraction from whole blood, washed mononuclear cells, and plasma from specimens collected in sodium heparin. These inhibitory effects on gene amplification could be reversed with heparinase. The addition of as little as 0.05 U of heparin completely inhibited amplification of an HLA-DQa sequence from placental DNA. We conclude that heparin can cause attenuation or inhibition of gene amplification. Acid citrate dextrose and EDTA, which lack inhibitory activity, are the most appropriate anticoagulants for clinical blood samples when polymerase chain reaction amplification is anticipated.
Holodniy
M
M
Center for AIDS Research, Stanford University Medical Center, California 94305.
Kim
S
S
Katzenstein
D
D
Konrad
M
M
Groves
E
E
Merigan
T C
TC
D019343:Citric Acid; D004279:DNA, Viral / Q000097:blood; D004492:Edetic Acid / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005784:Gene Amplification / Q000187:drug effects; D015750:Genes, gag; D005947:Glucose / Q000031:analogs & derivatives / Q000494:pharmacology; D006678:HIV / Q000187:drug effects* / Q000235:genetics; D015658:HIV Infections / Q000175:diagnosis; D006683:HLA-DQ Antigens / Q000235:genetics; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry / Q000382:microbiology; D010070:Oxalates / Q000494:pharmacology; D010920:Placenta / Q000737:chemistry; D016133:Polymerase Chain Reaction
D004279:DNA, Viral; D006683:HLA-DQ Antigens; D010070:Oxalates; C002113:acid citrate dextrose; D019343:Citric Acid; D006493:Heparin; D004492:Edetic Acid; D005947:Glucose