不同方法检测同种异基因造血干细胞移植受者巨细胞病毒感染的探讨.doc

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1、不同方法检测同种异基因造血干细胞移植受者巨细胞病毒感染的探讨作者：张龙天莉，周健，魏旭东，蒋东霞，程爱民，胡杰英，郭淑利，朱兴虎，宋永平【摘要】　　本研究探讨同种异基因造血干细胞移植(alloHSCT)后早期有效检测巨细胞病毒(CMV)感染的方法。应用荧光定量PCR和ELISA试剂盒分别检测19名alloHSCT受者，214份标本的血浆DNA负荷量和血清IgM抗体，同时应用流式细胞术检测188份标本白细胞pp65抗原。结果表明：pp65抗原、DNA定量和IgM抗体的阳性检出率分别为30.85%(58/188)、35.5

2、1%(76/214)和13.08%(28/214)，连续阳性病例和临床诊断的符合率分别为7/8、7/8和3/8。DNA定量与pp65抗原阳性检出率的差别无统计学意义(P＞0.05)，但两种检测方法有明显的相关性(P＜0.05)。IgM抗体阳性检出率明显低于DNA定量和pp65抗原，其差别均有统计学意义(P＜0.05)，与另两种检测方法虽有关系，但不密切。结论：流式细胞术和荧光定量PCR检测alloHSCT受者CMV早期感染可靠、简便快速，值得临床推广使用。【关键词】巨细胞病毒感染；同种异基因造血干细胞移植；流式细胞术；荧

3、光定量PCR；pp65抗原　　DetectionandScreeningforHumanCytomegalovirusInfectioninAllogeneicHematopoieticStemCellTransplantationRecipientsbyDifferentMethodsAbstractTheaimofstudywastoexplorethebetterdetectionmethodforcytomegalovirus(CMV)inallogeneichematopoieticstemcelltranspl

4、antation(alloHSCT)recipientsandtocomparetheefficiencyoffluorogenicquantitativePCR(FQPCR),flowcytometry(FCM)andELISA.TheplasmaDNAloadingandserumlevelofIgMantibodyagainstCMVin214clinicalspecimensfrom19alloHSCTpatientsweredetectedbyrealtimeFQPCRandELISArespectiv

5、ely,thepp65antigenin118peripheralbloodleukocytesamplesweremeasuredbyFCM.Theresultsshowedthatthepositiveratesofpp65antigen,IgMantibodyandDNAloadwere30.85%(58/188),13.08%(28/214)and35.51%(76/214)respectively,thecoincidencebetweentheirsequentialdetectionpositiverates

6、andclinicaldiagnosiswere7/8、7/8and3/8respectively.Therewasnostatisticalsignificantdifferencebetweenthepositiverateofpp65antigenandofDNAamount(P＞0.05),andtheyhavemanifestedrelationships(P＜0.05).ThepositiverateofIgMantibodydetectedbyELISAwasobviouslowerthanthatofDNA

7、quantitatedbyFQPCRandpp65antigendetected7byFCM,butthedifferencebetweenthemshowedstatisticalsignificance(P＜0.05),SmallerrelativitywasfoundbetweenIgMantibodydetectionandtheothertwomethods(P＞0.05).ItisconcludedthatFQPCRandFCMaresensitive,rapid,suitableandreliableme

8、thodsformonitoringrecipientreactiveCMVinfectionofalloHSCTrecipientsandareworthytoextensivelyuseforguidingantiviraltherapy.Keywordscytomegalovirusinfect