基于HLA组特异性单倍体全长测序方法对模棱两可结果的准确定型

发布时间：2018-06-02 02:30

本文选题：HLA抗原 + 基因　；参考：《中国组织工程研究》2017年20期

【摘要】：背景:传统的HLA测序分型技术都是对父系母系双倍体进行扩增,由于HLA多样性高,产生了大量的模棱两可难题,而采用基于组特异性单倍体全长测序分型方法来解决模棱两可结果的确认定型鲜有报道。目的:分析基于组特异性单倍体全长测序分型方法对HLA基因分型模棱两可结果准确定型的效果。方法:对2例模棱两可结果标本以低分辨率分型方法结果作为参考起点,通过组特异性扩增分离,基于等位基因全长单倍体的Sanger测序(PCR-SBT)分型。结果与结论:(1)2例模棱两可分型结果分别为A*02∶03∶01及C*07:02∶01∶01杂合新等位基因;(2)A*02∶03∶01杂合新等位基因与A*11∶01∶01∶01全长比较,在NT817由C发生突变为T;(3)C*07∶02∶01∶01杂合新等位基因与C*08∶01∶01比较,在NT879由A发生突变为G;(4)结果表明,以低分辨率分型方法结果作为参考起点,通过组特异性扩增分离,基于组特异性单倍体全长测序分型方法能准确定型HLA基因分型模棱两可结果并发现新等位基因。
[Abstract]:Background: traditional HLA sequencing techniques are used to amplify paternal diploidy. Due to the high diversity of HLA, there are a lot of ambiguous problems. However, it is rarely reported that the full length sequencing method based on group specific haploids is used to solve the ambiguous results. Objective: to analyze the effect of group specific haploid full-length sequencing method for accurate typing of HLA genotyping. Methods: the results of low resolution typing method were used as the reference starting point in 2 cases of ambiguous results. The allelic full-length haploid Sanger sequencing PCR-SBT typing was carried out by group specific amplification and isolation. Results and conclusions the results of two ambiguous genotyping cases were as follows: A02: 03: 01 and C7: 022: 01: 01 heterozygous new alleles: A0203: 01 heterozygous new alleles compared with A1101: 01: 01. The results showed that the new heterozygosity allele in NT817 was changed from C to TX 02: 01W 01. Compared with C0 08: 01: 01, the mutation in NT879 was from A to GN 4). The results showed that the results of low resolution typing method were used as the reference starting point and isolated by group specific amplification. Based on group specific haploid full-length sequencing method, HLA genotyping can be accurately identified and new alleles can be found.
【作者单位】：深圳市血液中心输血医学研究所;深圳市晋百慧生物有限公司;
【基金】：广东省医学科研基金(A2016222) 深圳市战略新兴产业发展专项资金(JSGG20160328103642937) 深圳市科技研发资金(JCYJ20160427172335974) 深圳市卫生计生系统科研项目(201401077)~~
【分类号】：R457.7

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