皖北地区1025例孕妇抗-HPA抗体的初步研究

Preliminary study on anti-HPA antibody in 1025 pregnant women in north of Anhui Province

目的:探讨皖北地区孕妇血小板抗体的产生与基因型分布情况,为中国孕妇是否进行产前血小板抗体筛查提供基础数据和统计学依据。方法:选择2016年12月至2017年12月在本院进行产检的1025例孕妇为研究对象,收集其外周血,用单克隆抗体特异性血小板抗原固定术对孕妇血清进行血小板抗体初筛,对于初筛阳性标本用二磷酸氯喹去除人类白细胞抗原(HLA)的作用后重复进行初筛实验,仍为阳性的标本利用抗原捕获酶联免疫吸附试验进行血小板抗体抗原特异性分析。对于抗-HLA抗体阳性标本用聚合酶链反应-序列特异性引物(PCR-SSP)法进行HLA基因型检测,对人类血小板抗原(抗-HPA)抗体阳性标本用PCR-SSP法进行HLA和HPA基因型检测。结果:1025例孕妇中血小板抗体阳性者共45例,总阳性率为4.39%,其中41例为抗-HLA抗体阳性,4例为抗-HLA抗体和抗-HPA抗体同时阳性;初次妊娠孕妇(410例)与多次妊娠孕妇(615例)比较,血小板抗体阳性者分别为6例和39例,阳性率分别为1.46%和6.34%,差异具有统计学意义(χ2=13.95,P<0.05);在多次妊娠孕妇中,有2次妊娠史(388例)、3次妊娠史(188例)、4次及以上妊娠史(89例)孕妇血小板抗体阳性者分别为18例、10例、11例,阳性率分别为4.64%、7.25%、12.36%,三组比较差异具有统计学意义(χ2=7.51,P<0.05)。结论:皖北地区孕妇血小板抗体阳性率为4.39%,其中抗-HLA抗体阳性率为4.39%,抗-HPA抗体阳性率为0.39%,且抗-HPA抗体阳性孕妇同时伴有抗-HLA抗体阳性;孕妇血小板抗体的产生与妊娠次数有着密切的关系,抗体阳性率随着妊娠次数的增加而增高;在皖北地区人群中致新生儿同种免疫性血小板减少症发生的致病抗体可能为抗-HPA-5b抗体;皖北地区血小板抗体阳性孕妇的基因分型中,频率较高的HLA-A特异性抗原有A2、A24、A11、A33,HLA-B特异性抗原有B46、B13、B60、B62、B58,HLA-DQ特异性抗原有DQ6、DQ2、DQ9、DQ5、DQ7,HLA-DR特异性抗原有DR9、DR4、DR15、DR7、DR12、DR13。

Objective:To investigate the production and genotype distribution of platelet antibodies in pregnant women in the north of Anhui Province,in order to provide basic data and statistical basis for prenatal platelet antibody screening in pregnant women in China.Methods:1025 pregnant women who underwent prenatal examination in our hospital from December 2016 to December 2017 were selected as subjects and their peripheral blood was collected.Screening platelet antibody of pregnant women serum by with Monoclonal antibody specific immobilization of platelet antigen at the beginning.Repeated screening experiment for positive cases with chloroquine phosphate to removal the role of human leukocyte antigen(HLA).Detect the platelet antibody antigen specific of the cases which was still positive by modified antigen capture ELISA.Detected the HLA genotype of anti-HLA antibody positive cases by polymerase chain reaction-sequence specific primer(PCR-SSP).Detected the HLA and human platelet alloantigen(HPA)genotypes of anti-HPA antibody positive cases by PCR-SSP.Results:There were 45 cases of platelet antibody positive in 1025 pregnant women,with a total positive rate of 4.39%,and 41 cases were anti-HLA antibody positive,and 4 cases were anti-HLA and HPA antibody positive.In first pregnancy pregnant women(410 cases)and multiple pregnancy pregnant women(615 cases),platelet antibody positive wemen was 6 cases and 39 cases,positive rate was 1.46%and 6.34%,the difference has statistical significance(χ2=13.95,P<0.05);in multiple pregnancy pregnant women,there were 2 pregnancies(388 cases),3 pregnancies(188 cases),4 pregnancies and above(89 cases),platelet antibody positive pregnant women was 18 cases,10 cases,11 cases,positive rate was 4.64%,7.25%,12.36%respectively,the difference in three groups had statistical significance(χ2=7.51,P<0.05).Conclusion:The positive rate of platelet antibody in pregnant women in the north of Anhui Province was 4.39%,of which the positive rate of anti-HLA antibody was 4.39%,the positive rate of anti-HPA antibody was 0.39%,and the anti-HPA antibody positive pregnant woman was accompanied with anti-HLA antibody.The occurrence of platelet antibody in pregnant women was closely related to the frequency of pregnancy,and the positive rate of the antibody increases as the frequency of pregnancies increases;the pathogenic antibodies causing NAIT may be anti-HPA-5b antibody in the north of Anhui Province;among the HLA-A,B,DQ,DR genotyping of anti-HLA antibodies positive women,HLA-A specific antigen sites with higher frequency is A2,A11,A24 and A33,HLA-B specific antigen sites with higher frequency is B46,B13,B60,B62 and B58,HLA-DR specific antigen sites with higher frequency is DQ6,DQ2,DQ9,DQ5 and DQ7,HLA-DQ specific antigen sites with higher frequency is DR9,DR4,DR15,DR7,DR13 and DR12.