摘要
目的研究95例血清学ABO亚型标本的分子机制。方法对95例血清学ABO正反定型不符亚型标本全部进行血清学确认和聚合酶链反应-序列特异性(PCR-SSP)ABO基因分型,对PCR-SSP无法检测到亚型等位基因的标本分别先后进行ABO基因第1~7外显子双链测序和单链测序,以确定突变位点以及突变点所在基因位置。结果95例标本共检测出34个ABO等位基因。其中5个常见ABO等位基因(ABO*A1.01、ABO*A1.02、ABO*B.01、ABO*O.01.01和ABO*O.01.02)和29个罕见亚型等位基因,包括16个ISBT命名等位基因(ABO*A2.01、ABO*A2.05、ABO*A2.13、ABO*A3.07、ABO*AW.37、ABO*AEL.05、ABO*B3.01、ABO*B3.05、ABO*BW.03、ABO*BW.07、ABO*BW.27、ABO*BEL.03、ABO*cisAB.01、ABO*cisAB.05、ABO*BA.02、ABO*BA.04)和5个dbRBC曾命名等位基因(A223、B309、Bw37、Bel09、Bw40)和8个均未命名新等位基因[ABO*B.01+978C>A、ABO*A1.02+248A>T、ABO*B.01+125dupT、ABO*B.01+(98+1G>A)、ABO*A1.02/ABO*B.01+1A>G,ABO*A1.02/ABO*O.01.01+28G>T,ABO*A1.02/ABO*B.01+538C>T,ABO*A1.02/ABO*O.01.01+797insT,后4个标本因标本量不足无法进行单链测序验证]。95例标本通过PCR-SSP确认亚型等位基因76例(21个ISBT和dbRBC已命名等位基因),剩余19例标本进行ABO基因第1~7外显子测序确认,其中确认8例未命名等位基因标本,剩余11例标本经测序未发现在检测范围内的亚型等位基因。结论研究揭示了95例血清学ABO亚型的分子遗传学背景,发现8种罕见的未命名新等位基因。血清学疑难血型的检测受到患者病理、生理等因素的干扰,ABO亚型鉴定需要血清学检测和基因检测相结合。
Objective To study the molecular mechanism of 95 samples of serological ABO subtypes.Methods A total of 95 samples with discrepancy between forward and reverse blood grouping were subjected to serological confirmation,and genotyped by polymerase chain reaction with sequence-specific primers(PCR-SSP).For those subtype alleles could not be detected by PCR-SSP,ABO gene exon 1-7 sequencing and gene single strand sequencing were performed successively to determine the mutation site and the gene location.Results A total of 34 ABO alleles were detected in 95 samples.Five common ABO alleles(ABO*A1.01,ABO*A1.02,ABO*B.01,ABO*O.01.01 and ABO*O.01.02)and 29 rare ABO alleles were identified,including 16 named alleles by ISBT(ABO*A2.01,ABO*A2.05,ABO*A2.13,ABO*A3.07,ABO*AW.37,ABO*AEL.05,ABO*B3.01,ABO*B3.05,ABO*BW.03,ABO*BW.07,ABO*BW.27,ABO*BEL.03,ABO*cisAB.01,ABO*cisAB.05,ABO*BA.02,ABO*BA.04)and 5 named alleles by dbRBC(A223,B309,Bw37,Bel09,Bw40)and eight unnamed alleles[ABO*B.01+978C>A,ABO*A1.02+248A>T,ABO*B.01+125dupT,ABO*B.01+(98+1G>A),ABO*A1.02/ABO*B.01+1A>G,ABO*A1.02/ABO*O.01.01+28G>T,ABO*A1.02/ABO*B.01+538C>T,ABO*A1.02/ABO*O.01.01+797insT].The last four samples could not be verified by single strand because of insufficient samples.In 95 samples,76 samples(21 named alleles of ISBT and dbRBC)were identified by PCR-SSP,and the remaining 19 samples were identified by exon 1-7 sequencing of ABO gene,of which 8 were identified as unnamed alleles,and the remaining 11 samples were not identified as subtype alleles.Conclusion The molecular genetic mechanism of 95 serological ABO subtypes was revealed,and 8 rare novel alleles were identified.The detection of ambiguous blood groups is influenced by factors such as patient pathology and physiology,therefore the combination of serological testing and genetic testing is suggested for the identification of ABO subtype.
作者
宿军
王夕钢
杨红霞
车玲玲
任甜甜
杨春晴
赵玲
王胜
SU Jun;WANG Xigang;YANG Hongxia;CHE Lingling;REN Tiantian;YANG Chunqing;ZHAO Ling;WANG Sheng(Weifang Central Blood Station,Weifang Blood Research Institute,Weifang 261043,China;Weifang People′s Hospital)
出处
《中国输血杂志》
CAS
2024年第2期145-150,共6页
Chinese Journal of Blood Transfusion
基金
潍坊市卫生健康委员会科研项目(WFWSJK-2020-131)。
作者简介
第一作者:宿军(1974-),男,副主任技师,Email:sjun0609@163.com;通信作者:王胜,Email:wfswsjws@163.com。
