PCR-SSP基因分型技术在Rh血型鉴定中的应用研究

Application of PCR-SSP genotyping in identification of Rh blood grouping

目的研究聚合酶链式反应-特异性序列引物(polymerase chain reaction-sequence specific primer,PCR-SSP)基因分型技术在Rh血型鉴定中的应用。方法收集广东省肇庆市无偿捐血者RhD阴性血型样本2例、RhD阳性样本1例,采用盐水法进行C、c、D、E、e抗原分型,对D抗原阴性的样本采用抗人球蛋白法进行确认。提取样本基因组DNA,并应用PCR-SSP基因分型技术特异性扩增RhD以及RhCE基因片段,并对D外显子进行检测。根据有无PCR产物以及产物长度,判断样本的基因型。结果 1例RhD阳性样本以及ccdee样本的血清学分型与基因分型结果相符合,1例Ccdee样本的血清学分型与基因分型结果不相符。结论 PCR-SSP基因分型技术在RhD血型鉴定中能识别血清意义上的假阴型,在临床输血实践中具有广阔的应用前景。

Objective To study the application of PCR-SSP genotyping technology in Rh blood group identification. Methods 2 Rh D negative cases and 1 Rh D positive case from donors in Zhaoqing city of Guangdong province were detected by series serological tests. The D antigen negative were confirmed by anti- human-globulin methods. Genomic DNA was extracted from each blood sample, RhD/CE genes and D exon were analyzed by polymerase chain reaction-sequence specific primer（PCR-SSP）. The genotypes of samples were identified by gel electrophoresis according to the length of PCR products. Results The serologic results of RhD-positive and ccdee samples were consistent with the genotyping. The Ccdee sample was mismatched. Conclusion PCR-SSP genotyping technology could recognize the ＂false＂ negative in RhD blood group identification.