抗原减弱AB亚型的血清学表型与基因检测

Serological phenotype and gene detection of antigen-weakened AB subtype

目的鉴定血清学正定型A、B抗原减弱或丢失的疑难血型。方法收集2021—2022年河南省人民医院A或B抗原减弱的标本,血清学常规采用ABO、RhD血型鉴定微住凝胶卡式法,基因检测采用序列特异性引物-聚合酶链式反应(PCR-SSP)和Sanger测序两种方法,扩增ABO等位基因1~7外显子。结果血清学方法8例标本正定型A或B抗原减弱或丢失,反定型为AB型。PCR-SSP法显示,8例标本均含有A和B抗原,基因分型为AB型。Sanger测序显示8例标本有7个共有的突变位点;与A1.01/B.01基因型相比,A1.02/B.01特异性突变位点为c.467C>T,A1.02/Bw.12特异性突变位点为c.278C>T、c.467C>T,A1.02/Bw.07特异性突变位点为c.467C>T、c.1055G>A,c.278C>T、c.467C>T引起编码的第93位、156位氨基酸由脯氨酸变成亮氨酸,c.1055G>A引起第352位氨基酸由精氨酸变成谷氨酰胺。8例标本中1例基因型为A1.01/B.01,1例基因型为A1.02/Bw.07,2例基因型为A1.02/Bw.12,4例基因型为A1.02/B.01。结论PCR-SSP基因型与Sanger测序基因型相一致,结合血清学实验结果,血型均为AB型。血清学方法结合基因检测能准确地鉴定血型,保证输血的安全性和有效性。

Objective To identify difficult blood groups with weakened or lost antigens A,B in forward grouping of serological test.Methods The samples with weakened A or B antigens were collected in Henan Provincial People's Hospital from 2021 to 2022.The microcolumn gel cassette method was routinely used for the serological identification of ABO,RhD blood groups.Polymerase chain reaction-sequence specific primer(PCR-SSP)and Sanger sequencing were used for gene detection.Exons one to seven of the ABO alleles were amplified.Results A or B antigens of eight specimens were identified as weakened or lost by the forward grouping of serological test and as AB blood group by the reverse typing.The PCR-SSP method showed that eight samples all contained A and B antigens,and the genotype was AB.Sanger sequencing showed 7 common mutation sites in the 8 samples.Compared with A1.01/B.01 genotype,the specific mutation site(s)was(were)c.467C>T for A1.02/B.01,c.278C>T and c.467C>T for A1.02/Bw.12 genotype,c.467C>T and c.1055G>A for A1.02/Bw.07.c.278C>T,c.467C>T changed the coded 93rd and 156th amino acid from proline to leucine,and c.1055G>A changed the 352th amino acid from arginine to glutamine.Among the eight specimens,there were one case of genotype A1.01/B.01,one of A1.02/Bw.07,two of A1.02/Bw.12,and four of A1.02/B.01.Conclusion The genotype detected by PCR-SSP and Sanger sequencing is consistent,and the blood types are all AB combined with serological results.Serological method combined with genetic testing can accurately identify blood type,and guarantee the safety and validity of blood transfusion.