Development and Evaluation of a Real-Time TaqMan-PCR for the Detection of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients

[摘要] Introduction:Cytomegalovirus(CMV)hasbeenrecognizedasthemostimportantviralpathogeninpersonsundergoingbonemarrowtransplantation(BMT).Inthisstudy,wepresentthedevelopmentofaTaqMan-basedreal-timePCRassaytoquantifyhumancytomegalovirus(CMV)DNAinperipheralbloodleukocytes(PBLs)ofbonemarrowtransplantationpatients.MaterialsandMethods:Aplasmidcontainingthetargetsequencefromthepp65region(UL83)ofCMVwasconstructedasapositivecontroltemplate.Serialdilutionsof107to101plasmidsperassaywereprepared.Peripheralbloodsampleswerecollectedfrompatientsaftertransplantation.CMVDNAwasquantifiedbyRQ-PCRinparallelwiththepp65antigenemiaassayinPBLsamples.Results:Thereal-timePCRassaycoulddetectCMVDNAinpatient'ssampleswithawidelinearrange,from10toover107copiesofCMV.Real-timePCRassayresultscorrelatedwiththoseoftheCMVpp65antigenemiaassay(P<0.0001).Discussion:TheTaqManassaymaybeausefultoolforrapidquantificationofCMVinfectionandformonitoringofCMVreactivationinbonemarrowtransplantationrecipients.Theresultsofbothquantitativeassaysweresignificantlycorrelated;however,theRQ-PCRassaywasmoresensitivethanthepp65antigenemiaassay.

[学科分类] 基础医学

[关键词] Cytomegalovirus;TaqMan PCR;pp65 Antigenemia;Bone Marrow Transplantation