多重PCR技术筛选Co^a、Lu^b、Yt^a、Kp^b阴性稀有血型方法的建立

Multiple PCR for Screening Co^a,Lu^b,Yt^a,Kp^b Negative Rare Blood Type

目的采用多重PCR筛选技术建立一个PCR反应体系,同时扩增Coa、Lub、Yta、Kpb阴性稀有血型,并应用于无偿献血人群稀有血型的筛选与建库。方法根据Coa、Lub、Yta、Kpb血型基因核心序列设计相应引物,采用多重PCR体系对献血者模版DNA进行扩增,对扩增产物进行电泳分析,根据是否扩增出对应片段图谱判断该稀有血型基因的存在。结果随机抽取500名献血者进行筛选鉴定,没有发现Coa、Lub、Yta、Kpb阴性的稀有血型。结论该方法能在一个反应体系内同时扩增4个稀有血型,具有准确、高效的优点,可在无偿献血人群稀有血型的筛选与建库工作中应用。

Objective To build up a PCR system to simultaneously amplify Co^a, Lu^b, Yt^a, Kp^b rare blood type by multiple PCR technique, the system can be applied to screen rare blood type among blood donors and set up database. Methods The corresponding primers according to the blood type gene core sequence of Co^a, Lu^b, Yt^a, Kp^b were designed. The PCR system （ PCR- SSP） that led by different primer sequence to amplify template DNA of blood donors was adopted, amplification products were analyzed by electrophoresis, according whether the corresponding rare blood type gene segments were amplified to judge the existence of rare blood type. Results 500 people who donate blood at our centre in June 2010 were randomly extracted, and rare blood type of Co^a, Lu^b, Yt^a, Kp^b negative was not discovered. Conclusion The method can be used to simuhaneously amplify four rare blood types in one reaction system and had the advantage of accuracy, efficiency and can be used to screen rare blood type among blood donors and set up database.