Watanabe G, Umetsu K, Yuasa I, Sato M, Sakabe M, Naito E, Yamanouchi H, Suzuki T
Department of Forensic Medicine, Yamagata University School of Medicine, Japan.
Electrophoresis. 2001 Feb;22(3):418-20. doi: 10.1002/1522-2683(200102)22:3<418::AID-ELPS418>3.0.CO;2-8.
We present a simple and rapid polymerase chain reaction (PCR)-based technique, termed consumed allele-specific primer analysis (CASPA), as a new strategy for single nucleotide polymorphism (SNP) analysis. The method involves the use of labeled allele-specific primers, differing in length, with several noncomplementary nucleotides added in the 5'-terminal region. After PCR amplification, the amounts of the remaining primers not incorporated into the PCR products are determined. Thus, nucleotide substitutions are identified by measuring the consumption of primers. In this study, the CASPA method was successfully applied to ABO genotyping. In the present method, the allele-specific primer only anneals with the target polymorphic site on the DNA, so it is not necessary to analyze the PCR products. Therefore, this method is only little affected by modification of the PCR products. The CASPA method is expected to be a useful tool for typing of SNPs.
我们提出了一种基于聚合酶链反应（PCR）的简单快速技术，称为消耗等位基因特异性引物分析（CASPA），作为单核苷酸多态性（SNP）分析的新策略。该方法涉及使用长度不同的标记等位基因特异性引物，并在5'末端区域添加几个非互补核苷酸。PCR扩增后，测定未掺入PCR产物中的剩余引物量。因此，通过测量引物的消耗量来鉴定核苷酸取代。在本研究中，CASPA方法成功应用于ABO基因分型。在本方法中，等位基因特异性引物仅与DNA上的目标多态性位点退火，因此无需分析PCR产物。因此，该方法受PCR产物修饰的影响很小。CASPA方法有望成为SNP分型的有用工具。
