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目的以核苷酸序列分析鉴定出血型血清学分型困难标本的ABO血型基因型。方法对常规血清学方法难以做ABO血型准确定型的3例被捡血样,采用PCR方法扩增其ABO等位基因的第6、7外显子,PCR产物采用直接测序和克隆测序的方法,以确定其基因型。结果 3例被检血样的血型血清学结果相似:红细胞与抗-A弱凝集,与抗-B强凝集,不与抗-A1凝集,其血清中存在抗-A,初步定型为AWB;经直接测序和克隆测序发现:3例被检血样都含有O等位基因,且它们的B等位基因在第7外显子都存在700C>G突变,证实其为B(A)02等位基因。结论核苷酸序列分析证实这3例血清学表现为AWB亚型个体均为B(A)表现型,基因型均为罕见的B(A)02/O型。
Objective To identify ABO blood group genotypes in patients with blood grouping type with nucleotide sequence analysis. Methods The blood samples were collected from 3 cases whose ABO blood type was difficult to be determined by conventional serological methods. The 6th and 7th exons of ABO allele were amplified by PCR. The PCR products were sequenced and sequenced directly Determine its genotype. Results The serological results of blood samples were similar in 3 cases: weak agglutination of erythrocytes and anti-A, strong agglutination with anti-B, no agglutination with anti-A1, anti-A in serum and AWB initial stereotype; Sequencing and cloning sequencing revealed that all the three blood samples tested contained the O allele, and their B allele had the 700C> G mutation in exon 7, confirming that it was the B (A) 02 allele. Conclusion Nucleotide sequence analysis confirmed that all three serotypes of AWB subtype were B (A) phenotype, and all genotypes were rare B (A) 02 / O.