作者单位:310003杭州浙江大学医学院附属儿童医院儿内科·感染性疾病·流式细胞术检测PP65抗原血症在诊断巨细胞病毒感染上的应用魏健汤永民郑季彦张晨美王燕儿沈红强钱柏芹【摘要】目的探讨流式细胞技术检测PP65抗原血症对巨细胞病毒(CMV)感染的诊断价值。方法对136例CMV感染患儿流式细胞技术检测血PP65抗原、实时荧光定量PCR(RFQ2PCR)检测尿CMVDNA及血清ELISA法检测CMVIgM,比较3种方法的检测结果,并以35例非CMV感染的患儿作为PP65检测的阴性对照。对18例PP65抗原阳性患儿进行短期随访。结果136例CMV感染患儿从血单个核细胞(MNC)中检出PP65抗原118例(118/136,阳性率8618%)。其中53例患儿分别作MNC和多形核白细胞(PMNL)检测,从MNC中检出PP65抗原45例(45/53,阳性率8419%),PMNL中检出PP65抗原43例(43/53,阳性率8111%),MNC与PMNL阳性率差异无统计学意义(χ2m=0125,P>0105)。PP65与尿CMVDNA2种方法所检测的结果间差异无统计学意义(χ2m=1178,P>0105);但PP65与血CMVIgM检测结果间差异有统计学意义(χ2m=52192,P<0101)。PP65抗原水平与CMV感染的病情程度有明显相关性,全身性感染通常伴高水平抗原血症,单脏器感染抗原水平相对较低或阴性(χ2=38151,P<01005)。PP65在感染恢复期转为阴性或低水平,病情恶化时明显增高。结论流式细胞技术检测PP65抗原血症是诊断活动性CMV感染的有效手段,量化结果有助于监测病情。【关键词】巨细胞病毒感染;抗原病毒;流式细胞术ApplicationofflowcytometrytodetectPP65antigenemiafordiagnosisandmonitoringofhumancytomegalovirusinfectionWEIJian,TANGYong2min,ZHENGJi2yan,ZHANGChen2mei,WANGYan2er,SHENHong2qiang,QIANBai2qin1TheChildren′sHospital,ZhejiangUniversitySchoolofMedicine,Hangzhou310003,China【Abstract】ObjectivesToevaluatetheclinicalsignificanceofflowcytometry(FCM)todetectthecytomegalovirus(CMV)PP65antigeninpatientswithCMVinfection1MethodsSamplesfrom35patientswithoutCMVinfectionwereusedasnegativecontrol1ThedefinitediagnosisofCMVinfectionwasbasedonthenationalcriteriaforCMVinfection1All136patientswithCMVinfectionwereexaminedwiththeFCMtodetectCMVPP65antigen,real2timefluorescencequantitative2polymerasechainreactionassay(RFQ2PCR)todetectCMV2DNAandELISAtomeasuretheserumlevelofIgMantibodyagainstCMV1Theresultsofthese3assaysin2groups(isolatedorganinvolvementanddisseminateddiseases)werecomparedandthesignificanceofPP65antigenemiawasevaluated1Ashort2termfollow2upwasundertakenin18patients1ResultsThepercentagesofPP65positivityinbloodmononuclearcells(MNC)andpolymorphicnuclearleukocyte(PMNL)from35negativecontrolpatientswere0121%±0109%witharangeof0-0141%and0124%±0110%witharangeof0112%20148%,respectively,whichwerenotsignificantlydifferent(t=01425,P>0105)1The95thpercentiles(P95)ofPP65inMNCandPMNLwere0139%and0145%,respectively,soacutoffvalueof≥0150%wasset1Ofthe136patientswithCMVinfection,118samplesfrom118patientswerepositiveforPP65antigenemiawithapositiverateof8618%,whichwasnotstatisticallydifferentfromthat(9014%,χ2=0191,P>0105)ofCMV2DNAdetectedbyRFQ2PCRassaybutitwassignificantlyhigherthanthat(4516%,χ2=51150,P<01005)ofthedetectionbyIgMmeasurement1PP65detectionwascorrelatedwithurineCMVDNAamplification(χ2=63178,P<0101)whilethedifferentdetectionratesbetweenthetwoassayswerenotstatisticallysignificant(χ2m=1178,P>0105)1PP65detectionwasnotcorrelatedwithserumIgMmeasurementwhilethedetectionratesbetweenthetwoweresignificantlydifferent(χ2m=52192,P<0101)1NosignificantdifferencewasfoundbetweenthedetectionratesofCMVinfectioninMNC(45/53,8419%)andPMNL(43/53,8111%)(χ2m=0125,P>0105)1HigherPP65antigenemialevelwascorrelatedwithsystemicCMVinfection,whilelowerlevelofPP65waseitherinthepatien...
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