Diagnosis of Indian Visceral Leishmaniasis by Nucleic Acid Detection Using PCR 英文参考文献.docVIP

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Diagnosis of Indian Visceral Leishmaniasis by Nucleic Acid Detection Using PCR 英文参考文献.doc

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Diagnosis of Indian Visceral Leishmaniasis by Nucleic Acid Detection Using PCR 英文参考文献

DiagnosisofIndianVisceralLeishmaniasisbyNucleic AcidDetectionUsingPCR PankajSrivastava,SanjanaMehrotra,PujaTiwary,JayaChakravarty,ShyamSundar* InfectiousDiseaseResearchLaboratory,DepartmentofMedicine,InstituteofMedicalSciences,BanarasHinduUniversity,Varanasi,India Abstract Background: PCR based diagnosisforVisceral Leishmaniasis(VL), despitenumerouspublished primers, remainsfar from beingappliedinthefield.ThepresentstudywasplannedtodesignaLeishmaniaspecificdiagnosticassayandtoevaluate itssensitivityandspecificityonasamplesize,whichtothebestofourknowledgeisthelargesteverscreenedinonestudy. Methods: Leishmania specific primers were developed using 18S rRNA gene and their sensitivity was evaluated on 500 parasitologically confirmed patients with VL and 25 Post Kala-azar Dermal Leishmaniasis (PKDL) patients. Specificity was calculated on 250 healthy endemic controls, 250 healthy non endemic controls and 250 non leishmanial diseases like malaria. Results:OurPCRassayhadasensitivityof87.8%(95%CI:84.1–89.8)using200mLofpatient’speripheral-blood.Specificity wasabsoluteinnon-endemichealthycontrolsandinsubjectswithdifferentdiseaseswhileinendemiccontrolsitwas84% (95%CI:78.9–88.0).Itsoverallspecificitywas94.6%(95%CI-92.8–96.1). Conclusions:ThePCRassaydevelopedissensitiveenoughtodetectthe18SrRNAgeneinanamountequivalenttoasingle parasiteorlessinaonemillionhumancellenvironment.ThehighsensitivityofthisPCRdiagnostictestwithrelativelynon- invasiveperipheralbloodsamplingmethodopensupthepossibilityofitsdeploymentinfieldfortheroutinediagnosisof VL. Citation:SrivastavaP,MehrotraS,TiwaryP,ChakravartyJ,SundarS(2011)DiagnosisofIndianVisceralLeishmaniasisbyNucleicAcidDetectionUsingPCR.PLoS ONE6(4):e19304.doi:10.1371/journal.pone.0019304 Editor:GeorgesSnounou,Universite′ PierreetMarieCurie,France ReceivedDecember29,2010;AcceptedMarch27,2011;PublishedApril29,2011 Copyright: ? 2011 Srivastava et al. This is an open-access article distributed under the terms of the Creative Commons Attribution