摘要
目的探讨山东省潍坊市RhD初筛结果呈阴性汉族无偿献血者的Rh血型表型及RHD基因多态性分布特点。方法选择2015年3月至2017年7月,于潍坊中心血站参加无偿献血的1307例RhD初筛结果呈阴性的汉族献血者为研究对象。其中,男性献血者为921例,女性为386例,年龄为18~55岁。采用血清学检测方法对1307例RhD初筛结果呈阴性献血者的RhD阴性进行确认。血清学检测采用3个不同厂家的抗-D血型定型试剂和间接抗人球蛋白试验。对经血清学检测确认为RhD阴性的献血者,进行Rh血型表型分型,并且计算各RhCcEe的表型频率和单倍型频率。选择2015年9月至2016年3月参加无偿献血的116例RhD初筛结果呈阴性献血者进行RHD基因检测。RHD基因检测采用PCR-序列特异性引物(SSP)法和人类红细胞RHD基因分型试剂盒,并且进行基因多态性分析。Rh血型表型频率和单倍型频率,采用方根法计算。各Rh血型表型频率分布的Hardy-Weinberg遗传平衡检验采用χ2检验。本研究遵循的程序符合2013年修订的《世界医学协会赫尔辛基宣言》要求,并且与所有献血者签署临床研究知情同意书。结果①本研究对1307例RhD初筛结果呈阴性献血者的RhD阴性进行确认的结果显示,RhD阴性献血者为1244例(95.2%),RhD变异型为63例(4.8%)。②本研究1244例RhD阴性献血者中,ccee、Ccee、ccEe、CCee、CcEe、CCEe表型献血者分别为773例(62.14%)、329例(26.45%)、83例(6.66%)、40例(3.22%)、18例(1.45%)、1例(0.08%)。cde、CdE、Cde、cdE单倍型频率分别为78.83%、0.06%、16.99%和4.12%。经Hardy-Weinberg遗传平衡检验,本研究纳入的1244例RhD阴性献血者的Rh血型表型频率观察值与期望值比较,差异无统计学意义(χ2=2.17,P>0.05)。③本研究116例RhD初筛结果呈阴性献血者的RHD基因分型结果显示,RHD外显子全部缺失基因型为77例(66.4%),RHD-CE(2-9)-D型为13例(11.2%),弱D15型为3例(2.6%),DEL RHD1227A纯合型为19例(16.4%),DEL RHD1227A杂合型为4例(3.5%)。结论本研究初步探明本地区RhD阴性献血者的Rh血型表型及RHD基因分布特点,可为确保本地区Rh阴性血型患者的临床输血安全,并为Rh阴性稀有血型库建立,提供参考依据。
Objective To explore characteristics of Rh blood group phenotypes and RHD gene polymorphism among Han nationality voluntary blood donors with RhD negative of primary screening in Weifang City,Shandong Province.Methods From March 2015 to July 2017,a total of 1307 Han nationality blood donors who were negative for RhD primary screening when participated in the voluntary blood donation at Weifang Blood Center,were selected as subjects.Among them,there were 921 male donors and 386 females,aged 18-55 years.Serological testing method was used to confirm RhD negative of 1307 blood donors with RhD negative of primary screening.Anti-D blood grouping reagents from 3 different manufacturers and direct antiglobulin test were performed for serological tests.Blood donors who were confirmed RhD negative by serological test were classified for Rh blood group phenotype,and the phenotype frequency of each RhCcEe and their haplotype frequencies were calculated.From September 2015 to March 2016,a total of 116 blood donors with RhD negative of primary screening were selected for RHD gene detection.PCR-sequence-specific primer(SSP)method and human erythrocyte RHD genotyping kit were used for RHD gene detection.And the gene polymorphism of RHD gene was analyzed.The frequencies of Rh blood group phenotypes and haplotypes were calculated by square root method.The test for Hardy-Weinberg equilibrium of the phenotype frequency distribution of Rh blood group phenotypes were conducted by chi-square test.The procedures followed in this study were accordance with the requirements of the World Medical Association Declaration of Helsinki revised in 2013,and all the blood donors signed the informed consents.Results①Among 1307 blood donors with RhD negative of primary screening from Weifang in this study,there were 1244 donors(95.2%)with RhD negative and 63 donors(4.8%)with RhD variants.②Among the 1244 RhD negative blood donors,there were 773 donors(62.14%)with ccee phenotypes,329 cases(26.45%)with Ccee,83 cases(6.66%)with ccEe,40 cases(3.22%)with CCee,18 cases(1.45%)with CcEe and 1 case(0.08%)with CCEe.The haplotype frequencies of cde,CdE,Cde and cdE were 78.83%,0.06%,16.99%and 4.12%,respectively.The result of test for Hardy-Weinberg equilibrium showed that there was no significant difference between the observed and expected values of Rh blood group phenotype frequencies in 1244 RhD negative blood donors(χ2=2.17,P>0.05).③Among 116 blood donors with RhD negative of primary screening,there were 77 cases(66.4%)with whole RHD exon deletion genotypes,13 cases(11.2%)with RHD-CE(2-9)-D,3 cases(2.6%)with weak D15,19 cases(16.4%)with homozygous DEL RHD1227A,and 4 cases(3.5%)with heterozygous DEL RHD1227A.Conclusions This study preliminarily investigate characteristics of Rh blood type phenotypes and RHD gene distribution among donors with RhD negative in Weifang City,which ensure the blood transfusion safety of RhD negative patients and provide scientific basis for the establishment of RhD negative rare blood group library.
作者
刘文迪
杨春晴
宿军
张建科
车玲玲
赵玲
Liu Wendi;Yang Chunqing;Su Jun;Zhang Jianke;Che Lingling;Zhao Ling(Department of Medical Laboratory Science,Weifang Medical University,Weifang 261042,Shangdong Province,China;Weifang Blood Center,Weifang Institute of Blood,Weifang 261043,Shandong Province,China)
出处
《国际输血及血液学杂志》
CAS
2020年第1期71-76,共6页
International Journal of Blood Transfusion and Hematology
基金
潍坊市卫生健康委员会科研项目(2017wsjs051)。
