建立多重PCR方法进行Vel-,Sc1-,GIL-及In^b-稀有血型检测

Multiplex PCR for screening Vel,Sc1,GIL,In^b negative rare blood type

目的建立稳定的多重PCR-SSP反应方法检测Vel-,Sc1-,GIL-及Inb-稀有血型,辅助鉴定稀有血型人士易产生的高频抗体。方法根据Vel,Scianna,Gill及Indian血型系统编码基因的单核苷酸多态性位点,设计特异性引物,对献血者DNA模板使用多重PCR方法进行检测,并使用PCR基因定点诱变技术和基因合成的方法制备出的Sc1,GIL,Inb及Vel阴性对照品作为对照,根据是否扩增出目标片段判断稀有血型基因的存在。直接测序验证此方法的准确性。结果建立了稳定的多重PCR-SSP反应体系,能够同时检测Vel-,Sc1-,GIL-及Inb-稀有血型,PCR-SSP结果与直接测序结果相一致。结论该方法能在同一体系中同时扩增4个稀有血型,具有快捷、高效且成本低的优点,为稀有血型人士用血及抗体特异性鉴定提供保障。

Objective To develop a multiplex PCR system for screening Vel, Scl, GIL, and Inb negative rare blood type, to improve the verification of the rare blood type and to identify the antibody to the high frequency antigen on erythroeyte. Methods Specifie primers were designed according to the moleeular mechanisms of Vel, Scianna, Gill and indian blood group antigens that have been clarified and the nucleotide mutation of the rare blood type. The control was produced by gene site-directed mutagenesis and gene synthesis technique. ~13ae multiplex PCR was used for blood group antigens genoty- ping, then the PCR results would indicate whether or not the patient is rare blood type. The aeeuracy of this method could be verified by sequencing. Results A multiplex PCR system for screening Vel, Scl, GIL, and Inb negative rare blood type in one PCR system was developed. The results of PCR-SSP can be verified by sequencing. Conclusion This method can screen four rare blood types in one PCR system at the same time. It is fast, high-throughput, and low cost, and improves the detection efficiency. More guarantee on blood transfusion can be provided to the patients with rare blood type and antibody specificity identifieation.