CAJ | 학술논문

目的建立可同时检测血型抗原Dib、k、Jsb1910、Jsb2019的一套稳定的基因分型方法,通过筛选获得所检测人群的稀有血型数据,可扩大中国稀有血型资料库.方法应用基于PCR的基因定点诱变技术制备Dib、k、Jsb1910、Jsb2019血型等位基因检测对照品.分别针对血型抗原Dib、k、Jsb1910、Jsb2019等位基因的单核苷酸多态性位点设计序列特异性引物,通过优化PCR条件建立多重PCR体系,并对4190份随机献血者样本进行Dib、k、Jsb1910、Jsb2019血型抗原基因分型.结果成功制备出Dib、k、Jsb1910、Jsb2019基因检测对照品,并成功构建检测血型抗原Dib、k、Jsb1910、Jsb2019的多重PCR体系,所建立的多重PCR体系具有良好的重复性和稳定性,4190份随机献血者样本中共检出2例Di(b-)样本,未检出k-和Js(b-)样本.结论多重PCR具有快捷、高通量且成本低的优点,可用于筛选稀有血型.获得的稀有血型可存入稀有血型数据库,为稀有血型患者及长期输血患者提供相配合的血液,减少输血不良反应的发生.
목적 건립가동시검측혈형항원Dib、k、Jsb1910、Jsb2019적일투은정적기인분형방법,통과사선획득소검측인군적희유혈형수거,가확대중국희유혈형자료고.방법 응용기우PCR적기인정점유변기술제비Dib、k、Jsb1910、Jsb2019혈형등위기인검측대조품.분별침대혈형항원Dib、k、Jsb1910、Jsb2019등위기인적단핵감산다태성위점설계서렬특이성인물,통과우화PCR조건건립다중PCR체계,병대4190빈수궤헌혈자양본진행Dib、k、Jsb1910、Jsb2019혈형항원기인분형.결과 성공제비출Dib、k、Jsb1910、Jsb2019기인검측대조품,병성공구건검측혈형항원Dib、k、Jsb1910、Jsb2019적다중PCR체계,소건립적다중PCR체계구유량호적중복성화은정성,4190빈수궤헌혈자양본중공검출2례Di(b-)양본,미검출k-화Js(b-)양본.결론 다중PCR구유쾌첩、고통량차성본저적우점,가용우사선희유혈형.획득적희유혈형가존입희유혈형수거고,위희유혈형환자급장기수혈환자제공상배합적혈액,감소수혈불량반응적발생.
Objective A reliable method for genotyping blood group antigens Dib,k,Jsb1910 and Jsb2019 was developed.Through screening for rare blood types,the National Rare Blood Bank of China may be enriched.Methods The controls for allele detection of blood groups Dib,k,Jsb1910 and Jsb2019 were prepared via polymerase chain reaction (PCR)-mediated gene site-directed mutagenesis (SDM) technique.Sequence-specific primers were designed according to known single nucleotide polymorphism (SNP) sites of alleles of blood groups antigens Dib,k,Jsb1910 and Jsb2019,a multiplex PCR system was developed by optimizing PCR reaction system.And 4190 random healthy donors samples were screened for the blood group antigens.Results Using SDM technique,controls for alleles in blood group Dib,k,Jsb1910 and Jsb2019 were successfully generated.And a multiplex PCR system for genotyping above blood groups was developed.After verification,the system has performed with good stability and reproducibility.Two Di (b-) samples have been discovered from 4190 samples,no k-and Js(b) sample was found.Conclusion Multiplex PCR features rapid detection,high throughput and low cost,and can be used for screening for donors of rare blood types.Information of donors may be registered in a database,which in turn can help those with rare blood types or require long-term blood transfusion to obtain matched blood,thereby reduce the adverse reactions of blood transfusion.