流式细胞术应用于Kidd血型Jka抗原检测的建立  

The Establishment and Application of Flow Cytometry on the Detection of Jka Antigen in Kidd Blood Group

作　　者：章昊[1] 梁爽 李乐兵 苏宇清[1] ZHANG Hao;LIANG Shuang;LI Le-Bing;SU Yu-Qing(Shenzhen Blood Center,Shenzhen 518035,Guangdong Province,China;School of Life and Marine Sciences,Shenzhen University,Shenzhen 518061,Guangdong Province,China)

机构地区：[1]深圳市血液中心,广东深圳518035 [2]深圳大学生命与海洋科学学院,广东深圳518061

出　　处：《中国实验血液学杂志》2022年第1期250-255,共6页Journal of Experimental Hematology

基　　金：深圳市科技计划项目(JCYJ20180302153621300);深圳市医疗卫生三名工程(SZSM2O1811092);深圳市输血医学重点学科(SZXK070);广东省医学基金(B2021093)。

摘　　要：目的:建立流式细胞术鉴定Kidd血型Jka抗原的实验方法,并验证其准确性。方法:随机选取96(人)份无偿献血者血液样本,将红细胞与IgG性质抗-Jka混合孵育,随后结合荧光素Alexa Fluor 647标记的羊抗人IgG二抗,采用流式细胞术检测样本红细胞表面Jka抗原。通过流式细胞术获得每个样品的荧光直方图。利用血清学凝集试验比较衡量流式细胞术检测Jka抗原强度的准确性,利用PCR-SSP与基因测序分型验证流式细胞术检测Jka抗原阴阳性的正确性。结果:流式细胞术对于红细胞Jka抗原强度的检测与血清学鉴定结果相一致,血清学凝集强度越高,流式细胞术检测荧光强度也越高,Jka抗原强度也越高。流式细胞术检测灵敏度优于血清学方法,Jka抗原阳性包括弱阳性样本与阴性样本的荧光强度差别很大,极易区分。全部样本流式细胞术检测结果与基因分型结果完全一致,证实了流式细胞术的准确性。结论:成功建立了流式细胞术鉴定红细胞Kidd血型Jka抗原的方法,灵敏度优于传统的血清学方法,可以准确区分不同强度的Jka血型抗原。Objective;To establish a based method flow cytometry to identify the antigen Jka in human red blood cells(RBCs) and verify its accuracy.Methods;A total of 96 blood samples were enrolled in the study randomly from the voluntary blood donors in Shenzhen Blood Center.The RBCs were incubated with IgG anti-Jka primary antibody,and then labeled with the secondary antibody antin-IgG-Alexa Fluor 647.The fluorescence histograms of each sample were obtained by flow cytometry.Serological agglutination test was used to compare the accuracy of flow cytometry in the detecting of antigen Jka,while PCR-SSP and gene sequencing genotyping were used to verify the accuracy of flow cytometry in the detecting of the antigen in human RBCs.Results:The results of flow cytometry for antigen Jka in human RBCs were consistent with those from serological tests.Samples that demonstrated higher serological agglutination intensity also showed higher fluorescence activity,which indicate more stronger of Jka antigen.The sensitivity of flow cytometry was higher than that of serological test;especially in distinguish Jka weak and negative samples.Flow cytometric results of all samples were consistent with the geno typing results,which confirmed the accuracy of flow cytometry.Conclusion:The study established a new flow cytometry-based method successfully for the identification of Jka antigen of Kidd blood group in human RBCs.The Kidd blood group antigen Jka of different intensities can be accurately distinguished by the technique.

关 键 词：KIDD血型 Jka抗原 流式细胞术 JKA基因 

分 类 号：R457.ll[医药卫生—治疗学]