等位基因频率在鉴别模棱两可HLA基因型中的应用价值分析  

Application Value of Allele Frequencies in Direct Identification of Ambiguous HLA Genotypes

作　　者：程良红[1] 邹红岩[1] 李桢[1] 金士正[1] 王大明[1] 高素青[1] 

机构地区：[1]深圳市血液中心组织配型与免疫遗传研究室,广东深圳518035

出　　处：《中国实验血液学杂志》2009年第2期487-492,共6页Journal of Experimental Hematology

基　　金：广东省科技计划项目(编号2008B030301278);广东省医学科研基金项目(编号A2008622);深圳市科技计划项目(编号200802117)

摘　　要：本研究探讨采用等位基因频率直接鉴别模棱两可HLA基因型的应用价值。应用聚合酶链式反应(PCR)-测序分型方法(SBT)对658名汉族供者HLA-A、B、DRB1基因进行测序分型,并分别采用PCR-序列特异性引物法(SSP)和杂合性歧义引物分离法(HAPRs)对其中的模棱两可标本进行精确分型,进而利用等位基因频率计算真基因型的相对概率并与真实结果进行比对。结果表明:222个HLA-A,238个HLA-B和107个HLA-DRB1基因的模棱两可标本中,真基因型的相对概率高于95%的比例分别为99.5%(221/222)、95.8%(228/238)和97.7%(104/107);与PCR-SSP和HAPRs分型结果相比,3个基因座的吻合率分别为100%(222/222)、99.6%(237/238)和99.1%(106/107),仅有B*3501/5501和DRB1*1201/1504不同,其相对概率值分别为40.3%和2.1%。结论:在大规模供者HLA基因的PCR-SBT分型中,应用等位基因频率直接鉴别模棱两可基因型的方法不仅具有较高的准确率和可信度,而且更加经济、简便、快捷,但在移植前供、患者HLA基因的确认试验中必须慎重应用。This study was aimed to investigate the application value of allele frequencies in direct identification of the ambiguous HLA genotypes. The HLA-A, HLA-B and HLADRB1 loci in 658 Chinese Han donor were detected by PCR- SBT method, the ambiguous genotyping samples were identified by using high resolution PCR-SSP and heterozygous ambiguity resolution primers （HAPRs） methods. The relative probability of true genotypes was calculated by using allele frequencies and was compared with true results. The results indicated that the relative probability of true genotype 〉 95% in 220 HLA-A ambiguous samples, 238 HLA-B ambiguous samples and 107 HLA-DRB1 ambiguous samples were 99. 5 % （ 221/222 ）, 95.8 % （ 228/238 ） and 97.7 % （ 104/107 ） respectively. As compared with phenotyping results detected by PCR-SSP and HARP methods, the matching ratios for HLA-A, HLA-B and HLA-DRB1 loci were 100% （222/ 222）, 99.6% （237/238） and 99.1% （106/107） respectively, while the mismatch genotypes were observed only in B ＊ 3501/5501 and DRB1 ＊ 1241/1504, the ralative probability of them were 40.3% and 2.1% respectively. It is concluded that the detection method using allele frequencies to directly identify the ambiguous HLA genotypes in large scale PCR-SBT genotyping of donors not only can give higher accurate and reliable results, but also is a simple, rapid and cost-saving method. This method has to be used with great care in the identify-test of patient-donor pair before the transplantation.

关 键 词：HLA基因型 模棱两可HLA基因型 等位基因频率 PCR-SBT 

分 类 号：R457.11[医药卫生—治疗学]