Inhibition of human immunodeficiency virus gene amplification by heparin.
Author(s): Holodniy M, Kim S, Katzenstein D, Konrad M, Groves E, Merigan TC
Publication: J Clin Microbiol, 1991, Vol. 29, Page 676-9
PubMed ID: 1909709 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effect of anticoagulant type on the amplification of Human immunodeficiency virus (HIV) DNA and RNA from whole blood, plasma, serum and peripheral blood mononuclear cells (PBMC).
Conclusion of Paper
Less amplification of DNA from whole blood and DNA and RNA from PBMC and plasma was observed when blood was collected into sodium heparin then when whole blood, PBMC, plasma or serum were collected in acid-citrate-dextrose (ACD), EDTA, potassium oxalate or without any anticoagulants. However, this effect could be reversed by incubation of the extracted DNA with heparinase.
Studies
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Study Purpose
The purpose of this study was to determine the effect of anticoagulant type on the amplification of HIV DNA and RNA from whole blood, plasma, serum and PBMC. Blood was obtained from 5 HIV positive patients and 3 HIV negative patients. HIV infected U1 cells were added to some blood specimens obtained from HIV negative patients. RNA was extracted using guanadinium thiocyanate-phenol-chloroform, and DNA was extracted using phenol-chloroform.
Summary of Findings:
Less amplification of DNA from whole blood and DNA and RNA from PBMC and plasma was observed when blood was collected into sodium heparin then when whole blood, PBMC, plasma or serum were collected in ACD, EDTA, potassium oxalate or without anticoagulants. However, this effect could be reversed by incubation of the extracted DNA with heparinase. Further, the authors show that the addition of ≥0.5 units/mL of heparin directly to the PCR reaction inhibited amplification of placental DNA, but that this effect was partially attenuated by the addition of 10-fold more Taq polymerase to the reaction or by incubation with heparinase.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Not specified
- AIDS/HIV-related
Platform:
Analyte Technology Platform RNA RT-PCR DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Blood and blood products Plasma
Serum
Whole blood
Peripheral blood mononuclear cells
RT-PCR Specific Targeted nucleic acid HIV gag
Biospecimen Acquisition Anticoagulant Acid-citrate-dextrose
Sodium heparin
Sodium EDTA
Potassium oxalate
None
PCR Specific Reaction solution Heparinase
No heparinase
PCR Specific Targeted nucleic acid HIV gag
Analyte Extraction and Purification Analyte purification Heparinase
No heparinase
PCR Specific Nucleic acid amplification 1 x Taq polymerase
10 x Taq polymerase