基于“血清学试验+分子生物学技术”对一例疑难血型的鉴定分析

Analysis of A Case of Difficult Blood Group Identification Based on“Serologic Tests+Molecular Biology Techniques”

目的通过分享一例ABO血型正反定型不符患者的血型鉴定分析过程,提高临床实验室对疑难血型的认识及处理能力,确保临床用血安全。方法采集贵阳市第二人民医院重症医学科一例ABO血型正反定型不符患者的血液标本,采用血型血清学试验和基因检测技术对患者ABO血型进行鉴定分析。结果患者样本ABO血型正定型为B型、反定型为AB型;不规则抗体筛查、直接抗人球蛋白试验和间接抗人球蛋白试验均为阴性,吸收放散试验提示红细胞上含有A抗原。PCR基因扩增及测序显示,样本基因与ABO★A1.02和ABO★B.01相符,基因型为A1/B型,表型为A1B型;ABO基因第6号外显子297A>G及第7号外显子467C>T、526C>G、657C>T、703G>A、796C>A、803G>C、874C>G、930G>A出现多个位点基因突变,其中7号外显子874C>G基因突变尚未记录在国际输血协会。结论通过血型血清学试验和分子生物学技术最终判定患者ABO血型为A1B型,患者7号外显子上的874C>G基因位点突变可能是本研究首次报道的一个新发现。基因位点突变干扰血型的正确鉴定,联合运用血清学试验和分子生物学技术,对于正确鉴定血型和保证临床安全输血治疗具有重要意义。

Objective To share the blood group identifi cation and analysis process of a patient whose ABO blood type does not match the positive and negative typing,so as to improve the recognition and processing ability of the clinical laboratory on diffi cult blood types and ensure the safety of clinical blood use.Methods The blood samples were collected from a patient whose ABO blood group did not match the positive and negative types in the Department of Critical Care Medicine of Guiyang Second Hospital.Serological test and gene detection were used to identify the ABO blood group of the patient.Results The positive type of ABO blood group in patient samples is B type and the negative type is AB type.Irregular antibody screening direct anti-human globulin test and indirect anti-human globulin test were negative.Absorption-emission test suggested that red blood cells contained A antigen PCR gene amplifi cation and sequencing showed that the genotype of ABO*A1.02 and ABO*B.01 was A1/B type and A1B type;ABO gene mutations in exon 6297A>G and exon 7467C>T,526C>G,657C>T,703G>A,796C>A,803G>C,874C>G,930G>A of the ABO gene have not been recorded in the International Blood Transfusion Association.Conclusion The ABO blood group of the patient was fi nally determined as A1B by serological test and molecular biological technology.The 874C>G gene mutation on the patient's exon 7 may be a new discovery revealed by this study for the fi rst time.Gene site mutation interferes with the correct identifi cation of blood type,and the combination of serological tests and molecular biology technology is of great signifi cance for the correct identifi cation of blood type and ensuring clinical safe blood transfusion treatment.