Lu Xiao-jun, Jia Yong-qian, Fan Hong, Zhang Lei, Jia Jin
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2007 Sep;38(5):882-4.
To study the efficiency of polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) in detecting the DNA base mutation and single nucleotide polymorphism (SNP).
By DNA sequencing and PCR-DGGE respectively, mutations in the D-loop of the mitochondrial genome were detected in patients with acute leukemia in period of diagnosis and remission, which was regarded as constitutional DNA. The comparison of results was made for evaluating above two methods. Also, the wild type DNA and mutational DNA were mixed at different proportions to mimic the presence of DNA polymorphism. PCR-DGGE was done to study its sensitivity for SNP.
The specificity and sensitivity of PCR-DGGE in detecting base mutation were 100% and 97.1% respectively. And PCR-DGGE could even detect the presence of polymorphism at rate of 0.5%.
PCR-DGGE has high sensitivity and specificity and can be applied to detection of DNA mutation and SNP, which is better method for checking DNA mutations.
研究聚合酶链反应-变性梯度凝胶电泳（PCR-DGGE）检测DNA碱基突变和单核苷酸多态性（SNP）的效率。
分别采用DNA测序和PCR-DGGE检测急性白血病患者诊断期和缓解期线粒体基因组D环区的突变，将其作为体质性DNA。对两种方法的结果进行比较以评估上述两种方法。此外，将野生型DNA和突变型DNA按不同比例混合以模拟DNA多态性的存在。进行PCR-DGGE以研究其对SNP的敏感性。
PCR-DGGE检测碱基突变的特异性和敏感性分别为100%和97.1%。并且PCR-DGGE甚至能够以0.5%的比例检测到多态性的存在。
PCR-DGGE具有高敏感性和特异性，可应用于DNA突变和SNP的检测，是检测DNA突变的较好方法。
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