Analysis of ABO serological phenotype and molecular biology results of 256 blood donors with ambiguous blood group / 中国输血杂志

ABSTRACT

【Objective】 To correctly identify the blood group of ABO and study its molecular biological characteristics. 【Methods】 Blood samples from blood donors with discrepancies in forward and reverse typing using the microplate method were subjected to both saline tube agglutination test for serological identification and polymerase chain reaction-sequence specific primers (PCR-SSP) for genotyping. Additionally, direct sequencing of exons 1 to 7 of the ABO gene was performed on some donor samples to analyze their blood phenotype, genotype and gene sequence. 【Results】 For 256 samples showing discrepancy between forward and reverse typing in microwell method, 119 were identified as normal ABO blood types, 90 were weakened ABO antibodies and 47 were ABO subtypes by serology tube test. According to the PCR-SSP genotyping test, 233 of 256 (91.02%) were consistent with serological phenotype and genotype, 17 of 256 (6.64%) were inconsistent, and 6 samples can′t read genotype based on the kit result typing table.A total of 17 genotypes were identified in 250 samples as AO1 in 56, AO2 in 58, AA in 50, BO1 in 31, BO2 in 17, BB in 8, O1O1 in 2, O1O2 in 7, AB in 13, AO4 in 1, A205O2 in 1, A205A in 1, A201A in 1, O1O4 in 1, O2O2 in 1, A201B in 1 and A205B in 1. Sequencing of exons 1 to 7 of the ABO gene was carried out in 78 samples, and 29 ABO alleles were detected, seven of which were common alleles (*A101, *A102, *A104, *B101, *O01, *O02, *O04), 22 of which were rare alleles (*A201, *A205, *Ax01, *Ax03, *Ax13, *Ax19, *Ax22, *Ael10, *B305, *Bel03, *Bel06/*Bx02, *Bw07, *Bw12, *Bw17, *Bw37, *O05, *O26, *O61, *B(A)04, *B(A)07, *cisAB01 and *cisAB01var). In addition, six rare allele mutation sites were identified (c.101A>G; c. 103_106delG; c. 146_147insGC; c. 259G>T; c. 322C>T; c. 932T>C). 【Conclusion】 The identification of ambiguous ABO blood group requires the combination of serological testing and molecular biological examination to correctly identify the blood type and ensure the safety of clinical blood transfusion.