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首页> 外文期刊>Journal of clinical laboratory analysis. >An improved PCR PCR ‐ CTPP CTPP assay for the detection of ADH ADH 1B Arg48His polymorphism
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An improved PCR PCR ‐ CTPP CTPP assay for the detection of ADH ADH 1B Arg48His polymorphism

机译：一种改进的PCR PCR - CTPP CTPP测定检测ADH ADH 1B Arg48His多态性

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Background ADH 1B Arg48His polymorphism is associated?with the development of alcohol‐related diseases. In this study, we aimed to explore an improved polymerase chain reaction with confronting two‐pair primers ( PCR ‐ CTPP ) assay for the detection of ADH 1B Arg48His polymorphism. Methods A mismatch was introduced at the 3′ end of each of the two allele‐specific to increase the specificity of the reaction. But beyond that, a new mismatch at‐3 positions of outer primers was designed to decrease the efficiency of the aforementioned primers and depresses the amplification of an internal nonspecific DNA control. A total of 180 samples from healthy volunteers Han Chinese were tested to evaluate this new assay. Results The protocol of PCR ‐ CTPP was successful for genotyping of ADH 1B Arg48His . The results from the improved PCR ‐ CTPP assay were confirmed by Sanger sequencing, and correct genotyping rates were 100%.The genotype frequencies were 49.44% (89 cases) for His/His, 46.67% (84 cases) for Arg/His, and 3.89% (seven cases) for Arg/Arg respectively. Conclusions This improved PCR ‐ CTPP assay is simple, rapid, cost‐effective, and reliable, specific for the detection of? ADH 1B Arg48His polymorphism in most clinical diagnostic laboratories.
机译：背景ADH 1B Arg48 His多态性有关？随着酒精相关疾病的发展。在这项研究中，我们旨在探讨改善的聚合酶链反应与对抗双对引物（PCR - CTPP）测定进行检测，用于检测ADH 1B Arg48His多态性。方法在两种等位基因特异性中的每种特异性的3'末端引入不匹配以增加反应的特异性。但是，除此之外，外引物的新失配旨在降低上述引物的效率，并抑制内部非特异性DNA对照的扩增。测试了来自健康志愿者的180个样本Han Chinese进行了评估这个新的测定。结果PCR - CTPP的方案成功用于ADH 1B Arg48His的基因分型。通过Sanger测序证实了改进的PCR-CTPP测定结果，并且正确的基因分率率为100％。基因型频率为他/他的46.67％（84例）的基因型频率为49.44％（89例），arg /他分别为3.89％（七种案件）分别为Arg / Arg。结论这种改善的PCR - CTPP测定简单，快速，经济高效，可靠，具体用于检测？ ADH 1B ARG48HIS多态性在大多数临床诊断实验室中。

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