HLA-Ⅱ血清学分型与微量SSP法基因分型的比较分析  

Comparative analysis of serologic typing and HLA-Ⅱtyping by micro-PCR-SSP

作　　者：武大林[1] 凌汉新[2] 丁红[1] 张怡[1] 

机构地区：[1]第一军医大学南方医院输血科外科中心实验室,广东广州510515 [2]广州市第十二人民医院肾内科,广东广州510620

出　　处：《第一军医大学学报》2002年第3期247-249,共3页Journal of First Military Medical University

基　　金：第一军医大学第一附属医院1998新技术攻关项目(98院字第011号)

摘　　要：目的验证微量序列特异性引物(SSP)技术进行HLA-Ⅱ类移植配型的准确性并探讨血清学分型错误发生的原因。方法采用聚合酶链反应结合微量SSP技术(简称微量PCR-SSP法)以及单克隆抗体血清学分型技术对血清样品进行HLA分型并比较其结果。结果(1)微量PCR-SSP法检测的110例样本中,99例检出HLA-DR的396个等位基因、11例检出HLA-DQ的22个等位基因,检出10%的单倍型个体;(2)两种方法对比研究发现单克隆抗体血清学分型检出错误或漏检率较高,其误差在DR和DQ分别为38.81%和50.75%;(3)错误发生和易混淆的抗原为:DR15/16、11/12、13/14、8、12和DQ5/6、8/9。结论微量SSP法可以准确检定血清学易漏检和发生错误的抗原等位基因。Objective To evaluatetheaccuracyof polymerasechainreactionwithsequencespecificprimers(PCR-SSP)in HLA-Ⅱgenotypingandanalyzethecausesof theerrorsoccurringduringthegenotyping.Method Bloodsampleswere obtainedformpatientswithchronicrenalinsufficiency,leukemiaor thalassemiaandalsofromnormalsubjects.HLA-DRand-DQgenotypingof theserafromthe110subjectswas performedusingmicro-PCR-SSPandcomparisonwas madewiththe resultsobtainedfrommonoclonalantibodyserologictyping.Result Of the110samplesdetectedby micro-PCR-SSP,396allelesof HLA-DRwereidentifiedin99casesand22of HLA-DQin11cases,and10%of thesubjectswereidentifiedas homozygoteindividuals.Examinationby bothof the2methodsin67casesindicatedhighratesof misseddiagnosesand misdiagnosesby serologictypingwiththe diagnosticdiscrepancyas highas38.81%and50.75%for HLA-DRand-DQ respectively.TheantigensDR15/16,11/12,13/14,8or12;DQ5/6,8/9wereamongthosethatfrequentlygaveriseto errors or confusion.Conclusion Micro-PCR-SSPmethodcanaccuratelydetecttheallelesof HLA-Ⅱantigensthatareeasyto be missedor mistakenby serologicaltypingmethod.

关 键 词：聚合酶链反应 序列特异性引物 人类白细胞抗原-Ⅱ类 等位基因分型 单克隆抗体 SSP法 移植配型 HLA-Ⅱ 基因分型 血清学分型 

分 类 号：R446.11[医药卫生—诊断学]