摘要
人类ＨＬＡ抗原分型在人类学、法医学、疾病易感性及器官组织移植等研究中有重要的意义。在骨髓移植中，进行供者、受者ＨＬＡ配型，选择相合的供体，减少ＧＶＨＤ发生，是移植成功的关键。目前国内主要采用血清学方法进行ＨＬＡ配型。由于方法上的缺点，常有约２０％左右的误差率，因此，基因分型技术的研究与应用势在必行。该研究通过设计合成ＨＬＡ基因序列特异性引物，建立了ＨＬＡ－ＤＲ基因分型的套式扩增和直接扩增ＰＣＲ－ＳＳＰ技术，并在骨髓移植配型中进行了应用。对两种分型技术进行了对比，两种方法各有特点，套式扩增在高度同源的ＤＮＡ序列扩增中有高度的特异性，并具有高度的灵敏性，很少量的ＤＮＡ即可满足ＰＣＲ扩增，但需两次扩增，非同源因素影响增大；直接扩增操作简便，一次扩增完成分型，但结构同源因素对特异性影响大。两种方法经周密设计均有高度的准确性。
HLA typing is significant in the study of anthropology, forensic medicine,disease susceptibility and organ transplantation. In bone marrow transplantation, the selection of matched donor is a key procedure for the engraftment of the transplant and for the reduction of degree of GVHD. Usually, serological and cellular methods are used for HLA typing in most clinical labs, but they have many methodological disadvantages. In this study, a nested PCR SSP and a direct amplification PCR SSP protocols for HLA DR genotyping were developed and were used in the selection of matched donor for sibling BMT. In comparison, the nested PCR SSP method was sensitive and accurate without the effect of homologous structures outside the HLA DRB1 exon 2 region,but two PCR reactions are needed while the direct amplification PCR SSP method was practical and accurate, but requires strict design of primers and PCR parameters. In conclusion, both of the two methods are rapid and accurate, but they have different characteristics in HLA genotyping.
出处
《军事医学科学院院刊》
CSCD
北大核心
1996年第4期269-273,共5页
Bulletin of the Academy of Military Medical Sciences
关键词
聚合酶链反应
HLA
抗原
DNA
基因
扩增
polymerase chain reaction
DNA polymerases
templates
HLA antigens
