Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion
Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion
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DOI：
10.1128/jcm.37.10.3402-3404.1999
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发表时间：
1999-10-01
影响因子：
9.4
通讯作者：
Okhravi, N
中科院分区：
文献类型：
--
作者：
Carroll, NM;Adamson, P;Okhravi, N
由于在Tag DNA聚合酶中存在细菌DNA而导致的假阳性的发生率是使用PCR诊断感染的障碍。我们描述了一种方法，该方法使用限制性内切酶来破坏污染序列作为嵌套PCR模板的能力，该PCR使用基于16S rRNA基因的引物。该方法在PCR扩增10 fg细菌DNA之前使用。这种方法可以很容易地适应临床应用所需的其他敏感pcr。
The incidence of false positives due to the presence of bacterial DNA in Tag DNA polymerase is an obstacle to the use of PCR in the diagnosis of infection. We describe a method that uses a restriction enzyme to destroy the ability of contaminating sequences to act as templates for a nested PCR which uses primers based on the 16S rRNA genes. The method was used prior to a PCR that amplified 10 fg of bacterial DNA. This method can be readily adapted to suit other sensitive PCRs required for clinical applications.