Evaluation of Type-Specific Real-Time PCR Assays Using the LightCycler and J.B.A.I.D.S. for Detection of Adenoviruses in Species HAdV-C 英文参考文献.docVIP

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Evaluation of Type-Specific Real-Time PCR Assays Using the LightCycler and J.B.A.I.D.S. for Detection of Adenoviruses in Species HAdV-C 英文参考文献.doc

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Evaluation of Type-Specific Real-Time PCR Assays Using the LightCycler and J.B.A.I.D.S. for Detection of Adenoviruses in Species HAdV-C 英文参考文献

EvaluationofType-SpecificReal-TimePCRAssaysUsing theLightCyclerandJ.B.A.I.D.S.forDetectionof AdenovirusesinSpeciesHAdV-C MorrisS.Jones1*,NolanRyanHudson2,CarlGibbins2,StephenL.Fischer3 1ViralandRickettsialDiseaseLaboratory,CaliforniaDepartmentofHealthServices,Richmond,California,UnitedStatesofAmerica,2ClinicalInvestigationFacility,David GrantMedicalCenter,TravisAirForceBase,California,UnitedStatesofAmerica,3NavalHospitalCampPendleton,CampPendleton,California,UnitedStatesofAmerica Abstract Sporadically,HAdVsfromspeciesHAdV-Caredetectedinacuterespiratorydiseaseoutbreaks.Torapidlytypetheseviruses, we designed real-time PCR assays that detect and discriminate between adenovirus types HAdV-C1, -C2, -C5, and -C6. SixteenclinicalisolatesfromtheCaliforniaDepartmentofPublicHealthwereusedtovalidatethenewassays.Type-specific TaqManreal-timePCRassaysweredesignedandusedindependentlytosuccessfullyidentify16representativespecimens. ThelowerlimitofdetectionforourLightCyclersingleplexreal-timePCRassayswerecalculatedtobe100,100,100,and50 genomic copies per reaction for HAdV-C1, HAdV-C2, HAdV-C5 and HAdV-C6, respectively. The results for the singleplex J.B.A.I.D.S.assaysweresimilar.Ourassaysdidnotcross-reactwithotheradenovirusesoutsideofspeciesHAdV-C,respiratory syncytialvirus,influenza,orrespiratorydiseasecausingbacteria.Theseassayshavethepotentialtobeusefulasdiagnostic toolsforspeciesHAdV-Cinfection. Citation:JonesMS,HudsonNR,GibbinsC,FischerSL(2011)EvaluationofType-SpecificReal-TimePCRAssaysUsingtheLightCyclerandJ.B.A.I.D.S.forDetection ofAdenovirusesinSpeciesHAdV-C.PLoSONE6(10):e26862.doi:10.1371/journal.pone.0026862 Editor:JianmingQiu,UniversityofKansasMedicalCenter,UnitedStatesofAmerica ReceivedJune10,2011;AcceptedOctober5,2011;PublishedOctober27,2011 Copyright: ? 2011 Jones et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalautho