摘要
目的对1例血清学血型鉴定困难的患者进行基因鉴定,分析引起血型鉴定困难的原因及突变基因。方法采用微柱凝胶法和试管法进行血清学血型鉴定、抗球蛋白试验和抗体筛查,采用聚合酶链反应(PCR)扩增结合测序及PCR产物克隆的方法,分析该患者ABO基因第6、7外显子区及其侧翼序列,寻找变异位点。结果患者血清学血型鉴定正反结果不一致,其红细胞与单克隆抗A抗体反应呈弱阳性(++),与单克隆抗B及抗H抗体反应均为强阳性(++++);其血清与A1型红细胞呈现弱凝集,但不凝集B和O型红细胞;患者直接及间接抗球蛋白试验均为阴性、血清抗体筛查为阴性。经测序发现,患者ABO基因存在多个变异,其第6外显子区存在261del和c.297A>G二处杂合变异,第7外显子区存在c.526C>G、c.640A>G、c.646T>A、c.657C>T、c.681G>A、c.703G>A、c.771C>T、c.796C>T、c.803G>C、c.829G>A、c.930G>A共11处杂合变异,以A101等位基因为标准序列进行比较,对照Blood Group Antigen Gene Mutation Database进行ABO等位基因突变分析,判断患者血型为B(A)04/O06血型。结论患者的B变异的基因型及弱AB的表型是引起血型鉴定正反不一致的主要原因,DNA基因分型有助于解决疑难样本的血型鉴定。
Objective To investigate the gene identification of a patient with serologic blood group discrepancy,and analyze the reason and variations. Methods Microcolumn agglutination and routine tube method were used for serologic blood group identification,and Coombs' and antibody screening were performed. Exon 6,7 and their adjacent intron region of ABO gene were amplified by polymerase chain reaction( PCR),and the PCR products were directly sequenced to identify the gene mutation. Results The forward and reverse tests of serologic blood group identification were not consistent. The red blood cells of the patient showed weak positive( + +) with monoclonal anti-A and positive( ++++) with monoclonal anti-B and anti-H. The patient's serum showed weak agglutination with type A1 red blood cells and no agglutination with type B and type O red blood cells. The direct and indirect Coombs and antibody screening were all negative. There were 2 heterozygous variations in exon 6( 261 del and c. 297 A > G) and 11 heterozygous variations in exon 7( c. 526 C > G,c. 640 A > G,c. 646 T > A,c. 657 C > T,c. 681 G > A,c. 703 G > A,c. 771 C > T,c. 796 C > T,c. 803 G > C,c. 829 G > A and c. 930 G > A) of ABO gene in the patient compared with the reference sequence of A101 allele. Based on the Blood Group Antigen Gene Mutation Database,B( A) 04 /O06 subgroup alleles were identified. Conclusions The variation of B allele and the weak AB phenotype are the main reasons of the discrepancy in blood group identification for the patient. DNA genotyping could help us with solving the blood group identification for complicated serologic discrepancy samples.
出处
《检验医学》
CAS
2014年第9期945-951,共7页
Laboratory Medicine
基金
上海市卫生局公共卫生优秀学科带头人培养计划(GWDTR201226)
