不同地区MNS血型系统与M、N、S、s、Mur+抗原基因型一致性分析

Consistency of genotyping and phenotyping in M,N,S,s, Mur+ antigen and MNS blood group system from multiple regions

目的探讨不同Mur表型人群在MNS血型系统上,应用不同方法和试剂检测结果的一致性。方法共检测2500人份的Mur抗原,采用血清学和荧光定量PCR方法,比较不同方法和试剂间检测结果的一致性。对所有Mur阳性血样和随机抽取的200例Mur阴性血样的M、N、S、s抗原,分别采用血清学和荧光定量PCR方法,探讨不同方法间结果的一致性。结果研究人群中通过血清学和基因分型方法,检测出Mur阳性个体119例(4.76%),Mur阴性个体2381例(95.24%),其中不同地区献血者中Mur阳性率分别为4.2%(渭南地区:42/1000)和4.6%(北京:23/500);不同地区三个月内无输血史的患者中Mur阳性率分别为3.8%(西安:19/500)和7%(佛山:35/500),三种方法经统计学检验一致性为100%(Kappa=1.000,P<0.05);对200例Mur阴性人群分别采用三个不同厂家单克隆抗-M、抗-N和MN基因型方法进行M、N抗原检测,其结果一致性分别为99.0%(Kappa=0.983),97.5%(Kappa=0.958),95.5%(Kappa=0.925);119例Mur阳性人群不同厂家单克隆抗-M、抗-N和基因分型检测结果的一致性分别为97.5%(Kappa=0.955),93.2%(Kappa=0.880),84.9%(Kappa=0.729);在S、s抗原检测方面,Mur阳性人群不同厂家单克隆抗体和基因型方法间检测结果一致性分别为97.5%(Kappa=0.736),93.3%(Kappa=0.625),41.2%(Kappa=0.057);Mur阳性人群不同厂家单克隆抗体和基因型方法间检测结果一致性分别为99.5%(Kappa=0.944),97.5%(Kappa=0.858),89.0%(Kappa=0.526)。结论不同方法和试剂对于Mur抗原的检测结果没有差异性,均可以作为Mur抗原的常规筛查方法;对于Mur阳性人群,在M、N、S、s抗原方面,常规应用单克隆抗体检测的血型血清学方法和荧光定量PCR基因分型方法,在结果上存在差异性,这提示我们MNSs抗原的正确定型需同时以Mur抗原的结果为参考。

Objective To evaluate the consistency of M,N,S,s antigen screening in multiple regions′of samples with positive and negative Mur,using monoclonal antibodies,polyclonal antibodies and genotyping.Methods Polyclonal anti-Mi^a,monoclonal anti-Mi^a,Mur genotyping through real-time PCR were used to screen for Mi^a antigen in erythrocyte.Mur positive samples and 200 randomly selected Mur negative samples were collected.Genotyping and serotyping of M,N,S and s antigen in these samples were carried out using monoclonal anti-Mi^a produced by I brand,and monoclonal anti-M,monoclonal anti-N,monoclonal anti-S and monoclonal anti-s produced by three different manufacturers.Results A total of 2500 samples were collected from three designated areas.The positive rate for donor were 4.2%(42/1000)in Weinan and 4.6%(23/500)in Beijing;In Cotrast to patient,the positive rate were 3.8%(19/500)in Xian and 7%(35/500)in Foshan.The consistency was 100%(Kappa=1.000,P<0.05)for Mur genotype,monoclonal anti-Mi^a and polyclonal anti-Mi^a.The consistency of monoclonal anti-M,monoclonal anti-N and MN genotype produced by three different manufacturers were 99.0%(Kappa=0.983),97.5%(Kappa=0.958)and 95.5%(Kappa=0.925),respectively,when tested with M and N antigen in Mur negative samples.When tested with M and N antigen in Mur positive samples,the consistency were 97.5%(Kappa=0.955),93.2%(Kappa=0.880),and 84.9%(Kappa=0.729),respectively,to the three antibodies.The consistency of monoclonal anti-S,monoclonal anti-S and Ss genotype produced by three different manufacturers were 99.5%(Kappa=0.944),97.5%(Kappa=0.858)and 89.0%(Kappa=0.526),respectively,when tested with Ss antigen in Mur negative samples.When tested with Ss antigen in Mur positive samples,the consistency was 97.5%(Kappa=0.736),93.3%(Kappa=0.625)and 41.2%(Kappa=0.057),respectively,to the three antibodies.Conclusion The consistency of monoclonal anti-Mi^a,polyclonal anti-Mi^a and Mur genotyping were 100%when used to screen for Mi^a antigen in erythrocyte,thus it is advisable to be used in those screenings.When tested with serotypes and genotypes of M,N,S and s antigen in Mur positive samples,the results showed inconsistency,thus it is advisable to carry out Mur antigen genotyping simultaneously with MNSs antigens genotyping to act as quality control.