摘要目的:探讨1个B(A)06亚型家系的血型血清学及分子遗传学特征。方法:选取2023年6月15日因"皮肤黄染、渐加重"于河南省儿童医院就诊，进行血型检测发现ABO正反定型不相符的1例新生儿高胆红素血症患儿为研究对象。采集该患儿及其家系成员(祖父、祖母、父亲、母亲和姑母)外周血各6 mL，使用血型血清学方法进行ABO血型鉴定，并使用核酸提取或纯化试剂BT-01提取人类基因组DNA，采用PCR法扩增 ABO基因第2~7外显子，将扩增成功的PCR特异性产物进行Sanger法测序，并以 ABO* A1.01等位基因序列为参考序列，分析患儿及其家系成员 ABO基因序列，确定 ABO基因型。本研究遵循的程序通过了河南省儿童医院伦理委员会的批准(批准号：2022-K-L036)。 结果:ABO血型血清学结果显示，患儿及其祖父、父亲、姑母均存在正反定型不符，患儿血型表型为A wB或B(A)，其祖父血型表型为A 2B或B(A)，其父亲和姑母血型表型为A 2B；其祖母和母亲正反定型相符，均为O型。患儿新生儿溶血病筛查试验结果显示，游离试验检出IgG类抗-A 1抗体，放散试验、直接抗人球蛋白试验及抗体筛查结果均呈阴性。Sanger测序结果显示，患儿存在c.261delG、c.297A>G、c.526C>G、c.657C>T、c.703G>A、c.796C>A和c.930G>A变异，患儿祖父存在c.297A>G、c.526C>G、c.657C>T、c.703G>A、c.796C>A、c.803G>C和c.930G>A变异，患儿祖母存在c.106G>T、c.188G>A、c.189C>T、c.220C>T、c.261delG、c.297A>G、c.646T>A、c.681G>A、c.771C>T和c.829G>A变异，患儿父亲和姑母存在c.106G>T、c.188G>A、c.189C>T、c.220C>T、c.261delG、c.297A>G、c.526C>G、c.646T>A、c.657C>T、c.681G>A、c.703G>A、c.771C>T、c.796C>A、c.829G>A和c.930G>A变异，患儿母亲存在c.106G>T、c.188G>A、c.189C>T、c.220C>T、c.261delG、c.297A>G、c.646T>A、c.681G>A、c.771C>T、c.829G>A变异。患儿 ABO基因型为 ABO* BA.06/ ABO* O.01.01，患儿祖父为 ABO* BA.06/ ABO* B.01，患儿祖母为 ABO* O.01.02/ ABO* O.01.02，患儿父亲和姑母为 ABO* BA.06/ ABO* O.01.02，患儿母亲为 ABO* O.01.01/ ABO* O.01.02，患儿及其祖父、父亲与姑母 ABO* BA.06等位基因是在 ABO* B.01等位基因的基础上第7外显子发生c.803C>G变异所致。 ABO* BA.06等位基因在该家系中稳定遗传。 结论:利用基因测序技术可对B(A)06亚型新生儿血型准确定型。新生儿ABO血型鉴定时，若正定型检测结果出现≤3＋凝集强度，应认真分析原因，必要时需使用分子生物学技术并结合家系基因测序结果共同判定。

abstractsObjective:To explore the serological and molecular genetic characteristics of a family with subtype B(A)06.Methods:A neonatal hyperbilirubinemia patient who was treated at Henan Children′s Hospital on June 15, 2023, due to " yellowing of the skin and gradual aggravation", and was found to have inconsistent ABO forward and reverse typing through blood type testing, was selected as the research subject. Six milliliters of peripheral blood were collected from the newborn and her family members (grandfather, grandmother, father, mother and aunt) respectively. ABO blood group identification was performed by the blood group serological method. Human genomic DNA was extracted using the nucleic acid extraction or purification reagent BT-01. ABO gene exons 2 to 7 were amplified by PCR. The PCR-specific products that were successfully amplified were sequenced by Sanger method. Taking ABO* A1.01 as the reference sequence, the ABO gene sequences of the newborn and her family members were analyzed to determine the ABO genotype. The procedures followed in this study were approved by the Ethics Committee of Henan Children′s Hospital (Ethics No. 2022-K-L036). Results:The serological results of ABO blood group showed that the newborn, her grandfather, father and aunt were all incompatible with the forward and reverse typing. The blood group phenotype of the newborn was A wB or B(A), the blood group phenotype of the grandfather was A 2B or B(A), the blood group phenotype of the father and aunt were A 2B, and the blood group phenotype of the grandmother and mother were both O. The screening test results of hemolytic disease of the newborn showed that the free test detected IgG anti-A 1 antibody, while the elution test, direct antiglobulin test and antibody screening results were all negative. The Sanger sequencing results showed that the newborn had variations of c. 261delG, c. 297A>G, c. 526C>G, c. 657C>T, c. 703G>A, c. 796C>A and c. 930G>A. Her grandfather had variations of c. 297A>G, C. 526C>G, c. 657C>T, c. 703G>A, c. 796C>A, c. 803G>C and c. 930G>A. Her grandmother had variations of c. 106G>T, c. 188G>A, c. 189C>T, c. 220C>T, c. 261delG, c. 297A>G, c. 646T>A, c. 681G>A, c.771C>T and c. 829G>A. Her father and aunt had variations of c. 106G>T, c. 188G>A, c. 189C>T, c. 220C>T, c. 261delG, c. 297A>G, c. 526C>G, c. 646T>A, c. 657C>T, c. 681G>A, c. 703G>A, c.771C>T, c. 796C>A, c. 829G>A and c. 930G>A. Her mother had variations of c. 106G>T, c. 188G>A, c. 189C>T, c. 220C>T, c. 261delG, c. 297A>G, c. 646T>A, c. 681G>A, c. 771C>T, and c.829G>A.The genotype of the newborn was ABO* BA.06/ ABO* O.01.01, her grandfather was ABO* BA.06/ ABO* B.01, her grandmother was ABO* O.01.02/ ABO* O.01.02, her father and aunt were ABO* BA.06/ ABO* O.01.02, and her mother was ABO* O.01.01/ ABO* O.01.02. The ABO* BA.06 allele of the newborn, grandfather, father and aunt was caused by the c. 803C>G variation in exon 7 based on the ABO* B.01 allele. The ABO* BA.06 allele can be stably inherited in this family. Conclusion:The blood type of neonatal patients with B(A)06 subtype can be accurately determined by gene sequencing technology. If the forward typing is ≤3+ agglutination intensity in newborn ABO blood group identification, the reason should be carefully analyzed, and the molecular biology technology and family gene sequencing results should be used to jointly determine if necessary.