滨海新区无偿献血者ABO亚型血清学特性及等位基因突变位点的分析

发布时间：2018-03-13 07:49

本文选题：ABO亚型　切入点：等位基因　出处：《天津医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过对滨海新区无偿献血人群ABO血型的血清学和分子生物学检测,掌握ABO亚型的血清学表现,并对ABO亚型等位基因突变位点进行分析。方法:使用ABO血型常规检测法对无偿献血者进行ABO血型初筛,发现正反定型不符的疑难血型;使用经典盐水试管法对正反定型不符标本进行ABO亚型鉴定;将血清学鉴定为ABO亚型的标本送往生工生物工程(上海)股份有限公司进行测序;对测序结果显示存在错义突变者进行蛋白质功能分析并使用PyMOL建立糖基转移酶空间模型。结果:45880名无偿献血者中初筛检测出正反定型不符标本,经进一步鉴定有20例符合ABO亚型血清学特征,分别为A2(1)、AX(2)、Bx(3)、Bm(1)、B3(1)、A2B(2)、AXB(4)、ABx(3)、B(A)(2),cisAB(1)。20例标本经过测序分析发现10个ABO等位基因错义突变,分别为1009AG、905 AG、940AG、503GT、28GA、588CG、550GA、700CG、640AG、803GC,并造成10个氨基酸置换,分别为R337G、D302G、K314E、R168L、G10R、C196W、V184M、P234A、M214V、G268A,对应的10个亚型等位基因分别为A205、AX18、AX13、Bw22、B310、Bx04、Bx08、B(A)02、B(A)04、cisAB01。错义突变导致的氨基酸置换在进化中为保守氨基酸,通过空间模型分析显示氨基酸置换前后其侧链的空间构象发生较大改变。结论:(1)ABO亚型在血清学检测中常出现异常减少和(或)增加的抗原和(或)抗体,造成血型检测正反定型不符,需掌握ABO亚型血清学特性以做出快速而准确的血型鉴定。(2)ABO等位基因由于发生错义突变而导致A或B糖基转移酶发生氨基酸置换,氨基酸置换后由于物理化学性质、带电性质和疏水性质以及氨基酸侧链间分子间作用力的改变而影响A或B糖基转移酶的催化活性和特异性,从而形成ABO亚型表型。
[Abstract]:Objective: to study the serological and molecular manifestations of ABO blood group in the population of free blood donation in Binhai New area, and to understand the serological manifestation of ABO subtype. The allelic mutation sites of ABO subtype were analyzed. Methods: the ABO blood group of unpaid blood donors was screened by routine detection of ABO blood group, and the difficult blood group was found to be not consistent with positive and negative typing. The ABO subtypes of positive and negative unmatched samples were identified by the classical salt water test tube method, and the samples identified as ABO subtypes were sent to bioengineering (Shanghai) Co., Ltd for sequencing. The results of sequencing showed that the missense mutant was used to analyze the protein function and the spatial model of glycosyltransferase was established by PyMOL. After further identification of 20 cases of ABO subtype serological characteristics, they were identified as A2G905 AX905 AG9AG905 AG905 AG905 AG905 AG503GT28GA588CG550GA550GA550GA700 CG640GCC, and 10 ABO allele missense mutations were found by sequencing analysis of 10 ABO allele missense mutations, respectively, after sequencing analysis, we found 10 ABO alleles missense mutations, respectively, which were AG905, AG905, AG905, AG905, AG503, GT28, AG503, GT288CG5, CG550GA550GA700, CG640GCC, and resulted in 10 amino acid replacements. The corresponding 10 alleles are A205AX18AX18AX13AX13B310Bx04BX010BX04BX04BX01. the amino acid replacement caused by missense mutation is conserved in evolution, and the corresponding 10 alleles are A205AX18AX18AX13AX13B310Bx04BX08BX08BX08BX02A02A04A01. the amino acid replacement caused by missense mutation is conserved in evolution, and the corresponding 10 alleles are A205AX18AX13AX13AX13B310Bx04BX08BX08BX08BX08BX02A02A01. The spatial model analysis showed that the spatial conformation of the side chain of Amino acid was changed greatly before and after amino acid replacement. Conclusion the abnormal reduction and / or increase of antigens and / or antibodies are often found in the serological examination of the ABO subtype. It is necessary to master the serological characteristics of ABO subtype in order to make rapid and accurate blood type identification. The missense mutation of the allele causes amino acid replacement of A or B glycosyltransferase. After the replacement of amino acids, the catalytic activity and specificity of A or B glycosyltransferases were affected by the changes of physicochemical, charged and hydrophobic properties and the intermolecular forces between amino acids, thus forming the phenotype of ABO subtype.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R457.11

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