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.
出处
《第一军医大学学报》
CSCD
北大核心
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)
