Analytical Performance of the Roche LightCycler？ Mycobacterium Detection Kit for the Diagnosis of Clinically Important Mycobacterial Species 英文参考文献.docVIP

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Analytical Performance of the Roche LightCycler？ Mycobacterium Detection Kit for the Diagnosis of Clinically Important Mycobacterial Species 英文参考文献.doc

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Analytical Performance of the Roche LightCycler？ Mycobacterium Detection Kit for the Diagnosis of Clinically Important Mycobacterial Species 英文参考文献

AnalyticalPerformanceoftheRocheLightCyclerH MycobacteriumDetectionKitfortheDiagnosisof ClinicallyImportantMycobacterialSpecies ShaheedV.Omar1*,AndreasRoth2,NazirA.Ismail1,LindaErasmus3,MarthieEhlers1,MarleenKock1, NuraanPaulse4,HalimaM.Said1,AnwarA.Hoosen1,UdoReischl5 1DepartmentofMedicalMicrobiology,UniversityofPretoria,Pretoria,SouthAfrica,2Laborfu¨rMolekulareDiagnostikundMikrobiologie,HELIOSKlinikumE.v.Behring, Berlin,Germany,3NationalTBReferenceLaboratory,NationalInstituteforCommunicableDiseases, Johannesburg, SouthAfrica, 4NationalHealthLaboratoryService, Kimberley,SouthAfrica,5Institutfu¨rMedizinischeMikrobiologieundHygiene,Universita¨tsklinikum Regensburg,Regensburg,Germany Abstract Background: The LightCyclerH Mycobacterium Detection Kit based on real-time PCR technology for the detection of Mycobacteriumtuberculosis,MycobacteriumaviumandMycobacteriumkansasiiwasrecentlydeveloped.Thisstudyevaluated itsanalyticalsensitivity,specificityandreproducibility. Methodology/PrincipalFindings:Plasmidstandardswerepreparedandusedtodeterminethelimitofdetection.Theassay wasalsoperformedagainstorganismsotherthanmycobacteria,othermycobacterialstrainsandinterferingsubstancesto exclude cross-reactivity and interference. Reference standards were prepared and tested to assess the assay’s reproducibility.AllPCRassayswereperformedusingtheLightCyclerH2.0Instrument.ThedetectionlimitforM.tuberculosis was 28 copies per microlitre. Neither cross-reactivity nor interference occurred with non-mycobacterial organisms and substancestested.Overallreproducibilityforconsecutivemeasurements,run-to-run,lot-to-lot,day-to-dayandlaboratory- to-laboratoryachievedacoefficientofvarianceoflessthantwopercent. Significance:TheLightCyclerHMycobacteriumDetectionkithasshowntobearobustandaccurateassaywiththepotential tobeusedasarapidTBdiagnostictest. Citation:OmarSV,RothA,IsmailNA,ErasmusL,EhlersM,etal.(2011)AnalyticalPerformanceoftheRocheLightCyclerHMycobacteriumDetectionKitforthe Diagnosiso