HLA-DQA反相杂交法分型与多聚酶链反应－单链构象多态性配型的比较  

Comparisonbetweentheresultsofnon┐radioactivereversedotblothybridizationandpolymerasechainreaction┐singlestrandedconformationpolymorphismmatchinginHLA┐DQAgenotyping

作　　者：李国选[1] 王申五[1] 于燕[1] 阎征 张玉琴 陆道培[1] 

机构地区：[1]北京医科大学人民医院血液病研究所

出　　处：《中华血液学杂志》1996年第1期23-25,共3页Chinese Journal of Hematology

摘　　要：为了验证多聚酶链反应－单链构象多态性方法在骨髓移植配型中的可靠性和准确性，采用掺入生物素的多聚酶链反应对ＨＬＡ－ＤＱＡ位点基因进行了特异性扩增。将所选区分ＨＬＡ－ＤＱＡ位点等位基因的特异性探针点于尼龙膜上，用标记的扩增产物与膜上探针杂交，并用碱性磷酸酶显色法检测杂交信号，确定待测标本的基因型。对２０对异基因骨髓移植的供、受者进行ＨＬＡ－ＤＱＡ位点的基因分型，并与多聚酶链反应－单链构象多态性配型作比较。结果表明，２０对供、受者的多聚酶链反应－单链构象多态性配型结果与该反相杂交法基因分型结果一致。多聚酶链反应－单链构象多态性方法是一种骨髓移植的供、受者配型。HLA-DQAaleleswereamplifiedbypolymerasechainreaction(PCR)withtheadditionofBio-11-dUTPinordertoverifythereliabilityandaccuracyofPCR-singlestrandedconformationpolymor-phism(PCR-SSCP)appliedtodonor-recipientmatchingforbonemarrowtransplantation(BMT).Ase-riesofsequence-specificoligonucleotideprobesdiferentiatingHLA-DQAalelesfromeachotherweredotedonthesamenylonfilterwithpositivecharge,andhybridizedwiththebiotinatedamplifiedprod-ucts.Hybridizationsignalsweredetectedwithalkalinephosphatasedevelopmentreaction.HLA-DQAgenotypingwasmadefor20leukemiapatientsandtheirsiblingsbeforeBMT,andtheresultswerecom-paredwiththeircorrespondentPCR-SSCPmatching.Itwasfoundthattheresultsofreversedotblothy-bridizationwereidenticaltothoseofPCR-SSCPmatchingforthe20pairsofdonor-recipient.ThePCR-SSCPprovedtobearapid,reliableandpracticalmatchingmethodforBMT.

关 键 词：骨髓移植 配型 HLA-DQA 反相斑点杂交 PCR 

分 类 号：R457.7[医药卫生—治疗学]