Open AccessEffects of heparin on polymerase chain reaction for blood white cells
Author(s) -
Yokota Masaharu,
Tatsumi Noriyuki,
Nathalang Oytip,
Yamada Tetsuaki,
Tsuda Izumi
Publication year - 1999
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/(sici)1098-2825(1999)13:3<133::aid-jcla8>3.0.co;2-0
Subject(s) - polymerase chain reaction , heparin , microbiology and biotechnology , real time polymerase chain reaction , dna , polymerase chain reaction optimization , hot start pcr , chemistry , polymerase , biology , gene , nested polymerase chain reaction , biochemistry
Polymerase chain reaction (PCR) has been used with increasing frequency to diagnose infectious and genetic diseases. In this study, the effects of heparin on PCR were investigated, because heparinized blood may sometimes be used in PCR studies. HLA‐DQA1 gene amplification was used as a model. PCR was clearly interfered with when heparinized blood was used as a source of template DNA, and the degree of interference was affected by the following three factors; (1) type of Taq DNA polymerase; (2) leukocyte count in blood; and (3) concentration of heparin contained. When additional tests were conducted with additions of definite heparin concentrations to a PCR reaction mixture, specimens with large amounts of DNA tended to exhibit less interference by heparin. The addition of ≥ 0.1 to 0.0016 U of heparin per reaction mixture (50 μl) suppressed DNA amplification in a dose‐dependent fashion. We therefore concluded that much care should be taken when heparinized blood is used as a PCR material. J. Clin. Lab. Anal. 13:133–140, 1999. © 1999 Wiley‐Liss, Inc.