Lee J C, Tsai L C, Chen C H, Chang J G
Department of Forensic Science, Central Police University, Taoyuan, Taiwan, Republic of China.
Forensic Sci Int. 1996 Oct 25;82(3):227-32. doi: 10.1016/s0379-0738(96)02003-8.
ABO blood groups were determined by the mutagenically separated polymerase chain reaction (MS-PCR). The products from two sets of PCR reactions using the same program for the nucleotides at positions 261 and 703 from cDNA at the ABO locus were used to distinguish A, B and O alleles. Two forward mutagenic allele-specific primers of different lengths for the ABO polymorphic site were paired with the same reverse primer in each PCR reaction. The 216 bp fragment of the PCR products for the 261th nucleotide was A or B allele-specific and the 195 bp fragment was O allele-specific. The 126 bp fragment of the PCR products for the 703th nucleotide was B allele-specific and the 106 bp fragment was A or O allele-specific. The ABO genotypes were determined by the intersection of the predicted alleles from these two PCR reactions. The PCR products were obtained using 10 ng of DNA in 50 microL of PCR reaction mixture, and electrophoresed in 4% agarose gel. In this study, 265 ABO-phenotype known samples (A: 31, B: 48, AB: 6 and O: 180) in Chinese were used. The results of ABO genotypes were AA: 1, AO: 30, BB: 2, BO: 46, AB: 6 and OO: 180. These results were confirmed by the PCR-RFLP ABO genotyping method. This technique is a simple, rapid, and reliable method for ABO genotyping.
ABO血型通过诱变分离聚合酶链反应（MS-PCR）来确定。使用相同程序对ABO基因座cDNA中第261位和第703位核苷酸进行两组PCR反应的产物，用于区分A、B和O等位基因。在每个PCR反应中，将两条针对ABO多态性位点的不同长度的正向诱变等位基因特异性引物与同一条反向引物配对。针对第261位核苷酸的PCR产物的216 bp片段是A或B等位基因特异性的，195 bp片段是O等位基因特异性的。针对第703位核苷酸的PCR产物的126 bp片段是B等位基因特异性的，106 bp片段是A或O等位基因特异性的。ABO基因型通过这两组PCR反应预测等位基因的交叉来确定。使用50 μL PCR反应混合物中的10 ng DNA获得PCR产物，并在4%琼脂糖凝胶中进行电泳。在本研究中，使用了265份已知ABO血型的中国样本（A：31份，B：48份，AB：6份，O：180份）。ABO基因型结果为AA：1份，AO：30份，BB：2份，BO：46份，AB：6份，OO：180份。这些结果通过PCR-RFLP ABO基因分型方法得到了证实。该技术是一种用于ABO基因分型的简单、快速且可靠的方法。
