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1、PCR检测儿童微小残留白血病的研究进展【摘要】 微小残留白血病(MRD)是指白血病经化疗缓解后在骨髓中仍存在形态上不能检测到的白血病细胞,是白血病复发的主要原因。儿童MRD的检测对判断儿童白血病的预后、制定白血病的治疗方案有重要意义。PCR方法在MRD检测中具有快速、特异、简便、经济和样本量少等特点。本文综述近年以多重PCR初筛白血病靶基因,以荧光定量PCR(FQPCR)追踪MRD方面所取得的进展。【关键词】微小残留白血病PCR多重PCRFQPCR儿童白血病 DetectionofMinimalRe
2、sidualDiseaseinChildrenLeukemiaPatientsbyUsingPCR——Revieiahasapotentialimportancetopredictclinicalouteandtomodifytreatmentprotocolsofthediseases.Althoughsomepatientsiahaveachievedpleteremissionaccordingtotheclinicalandmorphologicalcriteria,therearestillver
3、ylobersofmalignantcellsthatcannotbediscriminatedbymorphologyandremainedinbonemarroinimalresidualdisease(MRD)andisthemainreasonleadingtorelapse.MRDdetectionhasanimportantsignificancefordesigningtreatmentprotocols.SeveralmethodsofMRDdetectionhavebeendevelope
4、d.Theseincludeconventionalcytogeics,fluorescenceinsituhybridization(FISH),floetricimmunophenotyping(FCM),Southernblotandpolymerasechainreaction(PCR)techniques,etc.Eachofthesetechniqueshasitsadvantagesanddisadvantages,sonotallofthemaresuitableforclinicalMRD
5、detectionbecauseofseveralinherentdisadvantages,suchaslimitedsensitivity,timeconsuming,highcost,orrequiringhighqualityDNAorRNA.Forexample,thesensitivitiesofconventionalcytogeics,FISH,FCMandSouthernblotapproachesforMRDmonitoringare10-1-10-2,10-2,10-3-10-4a
6、nd10-1,respectively.Relatively,PCRcanreachagoodsensitivityof10-4-10-6,andshooreadvantages,suchasfast,specific,simpleandloinimalamountsofDNAorRNAfordetection,etc.,soPCRhasitsspecificfeaturesforMRDdetection.InthisrevieiaspecificmarkerbymuitiplexPCRandFQPCRi
7、nrecentyearsaresummarized.Keyinimalresidualdisease;PCR;multiplexPCR;FQPCR;childrenleukemiaJExpHematol2007;15(3):652-656微小残留白血病(minimalresidualdisease,MRD)是指白血病经化疗取得完全缓解后,骨髓中仍存在形态上不能检测到的白血病细胞,是引起白血病复发的主要原因。准确测定MRD对临床制定治疗方案具有重要意义。目前用于检测MRD的方法有核型分析、荧光原位杂交(fl
8、uorescentinsituhybridization,FISH)、流式细胞术(floetricimmunophenotyping,FCM)、Southernblot和PCR等,不同方法各有利弊。核型分析、FISH、FCM和Southernblot等的敏感性分别为10-1-10-2、10-2、10-3-10-4和10-1,难以满足对MRD诊断的需要,且都存在或操作费时,或费用较高,或对样本要求较高等不足之处。相