Evaluation and Validation of a Real-Time PCR Assay for Detection and Quantitation of Human Adenovirus 14 from Clinical Samples 英文参考文献.docVIP

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Evaluation and Validation of a Real-Time PCR Assay for Detection and Quantitation of Human Adenovirus 14 from Clinical Samples 英文参考文献.doc

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Evaluation and Validation of a Real-Time PCR Assay for Detection and Quantitation of Human Adenovirus 14 from Clinical Samples 英文参考文献

EvaluationandValidationofaReal-TimePCRAssayfor DetectionandQuantitationofHumanAdenovirus14 fromClinicalSamples DavidMetzgar1,GregSkochko2,CarlGibbins2,NolanHudson2,LisaLott3,MorrisS.Jones2* 1DepartmentofRespiratoryDiseaseResearch,NavalHealthResearchCenter,SanDiego,California,UnitedStatesofAmerica,2ClinicalInvestigationFacility,DavidGrant UnitedStatesAirForce(USAF)MedicalCenter,Travis,California,UnitedStatesofAmerica,3AdvancedDiagnosticLaboratory,LacklandAirForceBase(AFB),Texas,United StatesofAmerica Abstract In2007,theCentersforDiseaseControlandPrevention(CDC)reportedthatHumanadenovirustype14(HAdV-14)infected 106 military personnel and was responsible for the death of one U.S. soldier at Lackland Air Force Base in Texas. Identification of the responsible adenovirus, which had not previously been seen in North America and for which rapid diagnostictoolswereunavailable,requiredretrospectiveanalysisatreferencelaboratories.Initialquarantinemeasureswere alsoreliantonrelativelyslowtraditionalPCRanalysisatotherlocations.Toaddressthisproblem,wedevelopedareal-time PCRassaythatdetectsa225basepairsequenceintheHAdV-14ahexongene.Fifty-oneoropharyngealswabspecimens fromtheNavalHealthResearchCenter,SanDiego,CAandAdvancedDiagnosticLaboratory,LacklandAFB,TXwereusedto validatethenewassay.Thedescribedassaydetectedeightofeightand19of19confirmedHAdV-14aclinicalisolatesintwo separatecohortsfromrespiratorydiseaseoutbreaks.Thereal-timePCRassayhadawidedynamicrange,detectingfrom102 to107 copiesofgenomicDNAperreaction.Theassaydidnotcross-reactwithotheradenoviruses,influenza,respiratory syncytialvirus,orcommonrespiratorytractbacteria.Thedescribedassayiseasytouse,sensitiveandspecificforHAdV-14a inclinicalthroatswabspecimens,andveryrapidsinceturnaroundtimeislessthanfourhourstoobtainananswer. Citation:MetzgarD,SkochkoG,GibbinsC,HudsonN,LottL,etal.(2009)EvaluationandValidationofaReal-TimePCRAssayforDetectionandQuantitationof HumanAdenovirus14fromClinicalSamples.PLoSONE4(9):e7081.doi:10.1371/jour