發(fā)布時(shí)間：2018-03-13 07:49

  本文選題：ABO亞型　切入點(diǎn)：等位基因　出處：《天津醫(yī)科大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的:通過(guò)對(duì)濱海新區(qū)無(wú)償獻(xiàn)血人群ABO血型的血清學(xué)和分子生物學(xué)檢測(cè),掌握ABO亞型的血清學(xué)表現(xiàn),并對(duì)ABO亞型等位基因突變位點(diǎn)進(jìn)行分析。方法:使用ABO血型常規(guī)檢測(cè)法對(duì)無(wú)償獻(xiàn)血者進(jìn)行ABO血型初篩,發(fā)現(xiàn)正反定型不符的疑難血型;使用經(jīng)典鹽水試管法對(duì)正反定型不符標(biāo)本進(jìn)行ABO亞型鑒定;將血清學(xué)鑒定為ABO亞型的標(biāo)本送往生工生物工程(上海)股份有限公司進(jìn)行測(cè)序;對(duì)測(cè)序結(jié)果顯示存在錯(cuò)義突變者進(jìn)行蛋白質(zhì)功能分析并使用PyMOL建立糖基轉(zhuǎn)移酶空間模型。結(jié)果:45880名無(wú)償獻(xiàn)血者中初篩檢測(cè)出正反定型不符標(biāo)本,經(jīng)進(jìn)一步鑒定有20例符合ABO亞型血清學(xué)特征,分別為A2(1)、AX(2)、Bx(3)、Bm(1)、B3(1)、A2B(2)、AXB(4)、ABx(3)、B(A)(2),cisAB(1)。20例標(biāo)本經(jīng)過(guò)測(cè)序分析發(fā)現(xiàn)10個(gè)ABO等位基因錯(cuò)義突變,分別為1009AG、905 AG、940AG、503GT、28GA、588CG、550GA、700CG、640AG、803GC,并造成10個(gè)氨基酸置換,分別為R337G、D302G、K314E、R168L、G10R、C196W、V184M、P234A、M214V、G268A,對(duì)應(yīng)的10個(gè)亞型等位基因分別為A205、AX18、AX13、Bw22、B310、Bx04、Bx08、B(A)02、B(A)04、cisAB01。錯(cuò)義突變導(dǎo)致的氨基酸置換在進(jìn)化中為保守氨基酸,通過(guò)空間模型分析顯示氨基酸置換前后其側(cè)鏈的空間構(gòu)象發(fā)生較大改變。結(jié)論:(1)ABO亞型在血清學(xué)檢測(cè)中常出現(xiàn)異常減少和(或)增加的抗原和(或)抗體,造成血型檢測(cè)正反定型不符,需掌握ABO亞型血清學(xué)特性以做出快速而準(zhǔn)確的血型鑒定。(2)ABO等位基因由于發(fā)生錯(cuò)義突變而導(dǎo)致A或B糖基轉(zhuǎn)移酶發(fā)生氨基酸置換,氨基酸置換后由于物理化學(xué)性質(zhì)、帶電性質(zhì)和疏水性質(zhì)以及氨基酸側(cè)鏈間分子間作用力的改變而影響A或B糖基轉(zhuǎn)移酶的催化活性和特異性,從而形成ABO亞型表型。
[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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R457.11

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