沈阳地区20例ABO亚型的血清学和分子生物学研究及2例ABO新等位基因的认定

Serological and molecular study of 20 ABO subgroup samples in Shenyang, including identification of 2 ABO new alleles

目的研究沈阳地区20例ABO亚型标本的血清学特征和分子遗传背景。方法对20例ABO血型定型不一致的标本进行血清学检测、聚合酶链反应-序列特异性(PCR-SSP)基因分型、ABO基因的全编码区和内含子1转录因子结合位点测序。结果在20例标本中,共检出亚型10种(Am、Bw、Bx、B3、A;B、AxB、A;Bw、A;Bx、AwB和ABw),其中以AB亚型最多;共检测出ABO等位基因16个,其中常见ABO等位基因5个(A1.01、A1.02、B.01、O.01.01和O.01.02)、罕见等位基因9个(AW.37、BW.03、BW.08、B3.07、cisAB.02、cisAB.03、cisAB.06、BA.04和O.01.04)以及新等位基因2个(AM.03和cisAB.07);AM.03新等位基因与A1.02比对存在1个碱基c.912 C>A突变,导致304位丝氨酸变成精氨酸;cisAB07新等位基因与B1.01比对存在1个碱基突变,差异仅在797位碱基T>C错义突变,导致266位甲硫氨酸变成精氨酸,血清学发生改变,同时表达A和B抗原。结论研究揭示了20例ABO亚型的分子遗传学背景,并首次发现了2个新等位基因ABO;AM.03(c.912C>A突变)和ABO;cisAB.07(c.797T>C突变)。

Objective To study the serological characteristics and genetic background of 20 samples with ABO blood group discrepancies in Shenyang. Methods Serological test, polymerase chain reaction-sequence specific primer(PCR-SSP) and sequencing of the full coding of ABO gene and the Intron 1 were conducted in 20 samples with ABO blood group discrepancies. Results Ten subtypes(Am, Bw, Bx, B3,,A;B, AxB, A;Bw, A;Bx, AwB and ABw) were detected in 20 samples, with AB subtype as the dominant. Sixteen ABO alleles were found, including 5 common alleles(A1.01, A1.02, B.01, O.01.01 and O.01.02), nine rare alleles(AW.37, BW.03, BW.08, B3.07, cisAB.02, cisAB.03, cisAB.06, BA.04 and O.01.04) and two novel alleles(AM.03 and cisAB.07). The AM.03 allele had a nucleotide change at position 912(C to A) compared with A1.02 allele, which resulted in an amino acid substitution(S304 R).The cisAB.07 allele was observed a missense mutation at position 797(T to C) which resulted in an amino acid substitution(M266 R) compared with B.01 allele. The serologic had been changed, and both A antigen and B antigen were expressed. Conclusion The study revealed the genetic background of 20 samples with ABO blood group discrepancies, and two new alleles as ABO*AM.03(912 C to A) and ABO*cisAB.07(797 T to C) were first reported.