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首页> 外文期刊>Journal of clinical laboratory analysis. >Improved Allele-Specific PCR Technique for Kidd Blood Group Genotyping
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Improved Allele-Specific PCR Technique for Kidd Blood Group Genotyping

机译：改进的等位基因特异性PCR技术用于孩子血型基因分型

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Background: We developed an allele-specific polymerase chain reaction (AS-PCR) technique for Kidd blood group genotyping. Methods: Altogether, 340 blood samples from Thai blood donors at the National Blood Centre, Thai Red Cross Society, were tested with anti-Jka and anti-Jkb using the gel technique and the direct urea lysis test was used for screening Jk(a-b-) phenotype. For AS-PCR technique, different types of primers were used for JK*01 and JK*02 allele detections in known DNA controls. Results: Regarding JK*02 allele detection, the pseudopositve amplification products were found when using correctly matched forward primer and a single mismatch forward primer. Interestingly, one type of two mismatch pairing at the 3′ end of the forward primer can be used together with the newly designed reverse primer for Kidd blood group genotyping. It was found that the typing results in all samples obtained by serological techniques and newly developed AS-PCR technique were in agreement and this PCR technique also gave 100% concordance of results in 30 samples randomly tested twice and demonstrated reproducible results. Conclusion: This study shows that the in-house AS-PCR is simple, cost-effective, and convenient for Kidd blood group genotyping in routine laboratories, especially, in resolving serologic investigations.
机译：背景：我们开发了用于基德血型基因分型的等位基因特异性聚合酶链反应（AS-PCR）技术。方法：使用凝胶技术，对来自泰国红十字会国家血液中心泰国献血者的340份血液样本进行了抗Jka和抗Jkb的检测，并采用直接尿素裂解试验筛查了Jk（ab- ）表型。对于AS-PCR技术，在已知的DNA对照中，不同类型的引物用于JK * 01和JK * 02等位基因检测。结果：关于JK * 02等位基因检测，当使用正确匹配的正向引物和单个错配的正向引物时，发现伪正向扩增产物。有趣的是，可以将正向引物3'端的两种错配配对中的一种与新设计的反向引物一起用于Kidd血型基因分型。发现通过血清学技术和新开发的AS-PCR技术获得的所有样品的分型结果是一致的，并且该PCR技术还给出了在随机测试两次的30个样品中100％的结果一致性，并证明了可再现的结果。结论：这项研究表明，内部AS-PCR操作简单，经济高效，并且便于常规实验室中的Kidd血型基因分型，尤其是在解决血清学研究方面。

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