罕见P^(k)表型个体的血清学和分子生物学分析  

Serologic and molecular biology analysis of a rare P^(k) phenotype

作　　者：高欢欢[1] 张娜[1] 耿微[1] 孔凡生 GAO Huanhuan;ZHANG Na;GENG Wei;KONG Fansheng(Jining Blood Center,Jining 272000,China)

机构地区：[1]济宁市中心血站,山东济宁272000

出　　处：《中国输血杂志》2025年第3期426-430,454,共6页Chinese Journal of Blood Transfusion

基　　金：山东省医药卫生科技项目(202302071253);济宁市重点研发计划项目(2022YXNS154)。

摘　　要：目的 分析1例P^(k)表型个体的血清学特征及分子生物学结果。方法 选取2022年7月在济宁市中心血站进行意外抗体鉴定的1例P^(k)表型患者为研究对象。采用血清学方法鉴定血型和意外抗体。用PCR直接测序法对3-β-N-乙酰半乳糖氨基转移酶基因(B3GALNT1)和α-1,4-半乳糖基转移酶基因(A4GALT)编码区序列进行扩增和测序分析,并对B3GALNT1基因的2处突变位点进行单倍体序列分析。用PROVEAN、SIFT、PolyPhen2和Mutation Taster软件分析突变对蛋白的作用效应。结果 血清学特征提示患者为罕见P_2~k表型,体内含有抗-P抗体,其次子为P_2表型。测序结果显示患者B3GALNT1基因存在c.239T>A(p.Leu80Gln)和c.433C>T(p.Arg145Ter)复合杂合突变,进一步的单倍体分析显示其中1条单倍体存在c.239T>A,另1条单倍体存在c.433C>T,其次子携带c.433C>T(p.Arg145Ter)杂合突变;患者及其次子A4GALT基因均存在c.903C>G纯合突变。生物信息学预测c.239T>A和c.433C>T突变均对蛋白功能有害。结论 B3GALNT1基因c.239T>A(p.Leu80Gln)及c.433C>T(p.Arg145Ter)复合杂合突变可能是造成P^(k)表型的原因之一,其中c.239T>A突变未见报道。Objective To analyze the serological characteristics and molecular biology results for a P^(k) phenotype.Methods One patient with P^(k) phenotype upon unexpected antibodies at Jining Blood Center in July 2022 was selected as the study subject.The blood groups and unexpected antibodies of the proband and his second son were identified using sero-logical methods.The sequences of 3-β-N-acetylgalactosaminyltransferase gene(B3GALNT1)and the coding region of α-1,4-galactosyltransferase gene(A4GALT)were amplified and analyzed by PCR direct sequencing,and haploid sequence anal-ysis was carried out on the variant sites of the B3GALNT1 gene.PROVEAN,SIFT,PolyPhen2 and Mutation Taster were used to analyze the effect of mutations on the protein.Results Serological test results suggested that the proband was a P2k type,including anti-P antibody in his serum,and his son was the P2 phenotype.Sequencing analysis revealed that the proband had a compound heterozygous variants of the B3GALNT1 in c.239T>A(p.Leu80Gln)and c.433C>T(p.Arg145Ter).Further haploid analysis showed that the c.239T>A had occurred in one haploid while c.433C>T was present in the other haploid,and his son carried a heterozygous variant of c.433C>T(p.Arg145Ter);the proband and his son all have homozygous mutation variants of the A4GALT in c.903C>G.Bioinformatic analysis also predicted that the mutations were harmful to the protein function.Conclusion The compound heterozygous variants c.239T>A(p.Leu80Gln)and c.433C>T(p.Arg145Ter)in the B3GALNT1 gene may contribute to the P^(k) phenotype,of which the c.239T>A variant has not been reported.

关 键 词：P^(k)表型 3-β-N-乙酰半乳糖氨基转移酶 基因测序 

分 类 号：R457.11[医药卫生—治疗学]