摘要
目的探讨RhD变异型DⅥⅢ型的分子生物学机制。方法选取2020年11月至2022年5月北京市红十字血液中心血型室接收的RhD血型弱阳性患者血样。使用3个厂家的单克隆抗-D(1个IgM、2个IgM/IgG抗-D)检测患者RhD血型,其中IgM抗-D采用盐水试管法(saline tube method,S)、IgG抗-D采用间接抗人球蛋白试验(indirect anti-human globulin test,IAT);采用直接抗人球蛋白分型卡和Rh分型卡检测患者直抗(direct anti-human globulin test,DAT)和RhCcEe抗原。采用实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测RHD基因缺失情况、RHD基因10个外显子全序列及其相邻侧翼内含子测序分析样本RHD基因型。结果共纳入RhD血型弱阳性患者血样8份,盐水试管法均只与一个厂家的抗-D试剂(IgM/IgG抗-D)呈弱阳性反应,IgM抗-D和另一家IgM/IgG抗-D阴性;IAT与2个厂家的抗-D试剂(IgM/IgG抗-D)均4+;DAT均阴性;RhCcEe表型分别为Ccee(6/8)、ccee(1/8)、CcEe(1/8)。实时荧光定量PCR结果均为RHD基因外显子E3~E6缺失,RHD基因外显子全序列测序均为RHD^(*)06.03.01。结论通过分子生物学方法对RHD基因的检测能准确鉴定为DⅥⅢ型,为受血者和供血者在临床输血中提供不同的保障策略。
Objective To explore the molecular biological mechanism of RhD variant DⅥⅢ.Methods The blood samples of patients with weak positive RhD blood group received by the blood group room of Beijing Red Cross Blood Center from November 2020 to May 2022 were selected.Three batches of monoclonal anti-D reagents(one batch of IgM and two batches of IgM/IgG anti-D)were used to detect IgM anti-D by saline tube method,and IgG anti-D by indirect antihuman globulin test(IAT).The anti-human globulin screening card and Rh typing card were used for direct anti-human globulin test(DAT)and RhCcEe antigens detection respectively.RHD gene deletions were identified by real-time flurescence quantitative polymerase chain reaction(PCR)and RHD genotypes were subsequently analyzed through genetic sequencing of 10 exons and adjacent flanking introns.Results A total of eight blood samples of patients with weakly positive RhD blood group were included,and all of them were weakly positive only with one batch of monoclonal anti-D reagent(IgM/IgG anti-D)and negative with the others.IAT results were 4+with two batches of anti-D reagents(IgM/IgG anti-D)and DAT results were negative.RhCcEe phenotype were:Ccee(6/8),ccee(1/8),CcEe(1/8).RHD exon E3-E6 deletion was found in all patients by real-time PCR.All exons of RHD gene were sequenced as RHD^(*)06.03.01.Conclusions RHD gene is accurately identified as DⅥⅢtype by molecular biology method,which provides different guarantee strategies for blood recipients and donors in clinicalblood transfusion.
作者
张烨
李晓菲
刘婷婷
许志远
郑仲男
Zhang Ye;Li Xiaofei;Liu Tingting;Xu Zhiyuan;Zheng Zhongnan(Department of Blood Group Section,Beijing Red Cross Blood Center,Beijing 100088,China)
出处
《北京医学》
CAS
2023年第10期913-916,共4页
Beijing Medical Journal
