MN血型四种方法检测比较分析  

Comparative analysis of four kinds of methods to detect the MN blood group

作　　者：许德义 孙春霞[2] 高家良 邓刚 梁伟 贺云蕾 彭明喜 童微星[4] 

机构地区：[1]宁波市中心血站输血研究所,浙江宁波315041 [2]宁波大学附属医院,浙江宁波315020 [3]鄞州人民医院,浙江宁波315040 [4]浙江大学宁波理工学院,浙江宁波315100

出　　处：《中国优生与遗传杂志》2015年第8期37-39,4,F0002,共5页Chinese Journal of Birth Health & Heredity

基　　金：宁波市医学科技计划项目(编号:2011A06)

摘　　要：目的比较MN血型四种方法检测方法的差异,为MN血型检测的准确性研究提供参考。方法分别用血型血清学、免疫印迹法、PCR-SSP法、直接测序法检测MN血型。血型血清学以免疫学为基础,按抗原抗体反应原理测定;免疫印迹法是把SDS电泳分离的红细胞膜蛋白从凝胶转移到PVDF膜上,并检测目标蛋白。PCR-SSP法针对红细胞MN血型系统基因不同碱基的变异设计特异性引物进行测定;直接测序法可测出MN血型的相关外显子基因序列,并与正常人相应的测序进行比对。结果血清学方法检测MN血型200例,其中MM 57例(28.5%),MN 87例(43.5%),NN 56例(28%);基因频率:m为50.25%,n为49.75%。Ss血型系统中,表现型为ss 198例、Ss 2例,未检到SS血型。免疫印迹法结果表明不同血型的红细胞之间膜蛋白图谱基本相似,但存在一些差异点,提示不同血型的红细胞之间可能存在膜蛋白的差异。PCR-SSP法和测序法解决了血清学方法抗体特异性、大量输血患者或自身溶血性贫血患者等问题。4种方法测得的血型结果一致,符合率100%。其中直接测序法可获得核苷酸的变异,使得该方法在研究血型遗传机制中发挥着重要的作用。结论 4种方法检测结果之间不存在差异,但均有一定的局限性和优势,可实现相互验证,确保MN血型检测的准确性。Objective：To compare the differences between four methods about detecting MN blood group to provide reference for the study of accuracy of MN blood group detection. Methods：Using serologic method,Western blot,PCR-SSP method,direct DNA sequencing to detect MN blood group,serologic method based on immunology,according to the principle of antigen antibody reaction. Western blotting is a technique that electrophoretic transfer of membrane proteins from sodium dodecyl ulfatepolyacrylamide gel electrophoresis（SDS-PAGE）gels to polyvinylidenefluoride（PVDF）membrane and detecting a protein target. PCR-SSP is a method that detect Specific primers designed according to the variation bases of MN blood group system gene. Direct DNA sequencing method can detect exon sequences of MN blood group,and sequences were compared with the normal people. Results：The MN blood group-associated variants were determined by serologic method,200 Chinese subjects studied,MM was present in 57（28.5%）of 200,MN was present in 87（43.5%）of 200,NN was present in 56（28%）of 200;Gene frequency：m 50.25%,n 49.75%. And Ss blood group system,ss was found in 198 of 200,Ss was found in 2 of 200,SS had not been detected. Western blotting results show that different red cell membrane proteins about blood group were similar,but there were some differences. Differences may exist between membrane proteins in different blood red cell. PCR-SSP and DNA sequencing method have solved the inadequate in serological antibody specificity,massive transfusion or autoimmune hemolytic anemia in patients. The results of 4 methods measured blood group were the same,the coincidence rate was 100%.The direct DNA sequencing method could obtain nucleotide variations,the method plays an important role in the study of genetic mechanism of blood group. Conclusion：There were no difference between the results of the 4 methods,but there were some limitations and advantages,they can realize mutual authentication,to ensure the accuracy of MN blood group detection

关 键 词：MN血型 免疫印迹法 PCR-SSP GYPA基因 DNA测序 

分 类 号：R446.6[医药卫生—诊断学]