A Novel High Throughput Assay for Anthelmintic Drug Screening and Resistance Diagnosis by Real-Time Monitoring of Parasite Motility 英文参考文献.docVIP

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A Novel High Throughput Assay for Anthelmintic Drug Screening and Resistance Diagnosis by Real-Time Monitoring of Parasite Motility 英文参考文献.doc

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A Novel High Throughput Assay for Anthelmintic Drug Screening and Resistance Diagnosis by Real-Time Monitoring of Parasite Motility 英文参考文献

ANovelHighThroughputAssayforAnthelminticDrug ScreeningandResistanceDiagnosisbyReal-Time MonitoringofParasiteMotility MichaelJ.Smout1,AndrewC.Kotze2,JamesS.McCarthy3,AlexLoukas1* 1JamesCookUniversity,Cairns,Australia,2CSIROLivestockIndustries,Brisbane,Australia,3QueenslandInstituteofMedicalResearch,Brisbane,Australia Abstract Background: Helminth parasites cause untold morbidity and mortality to billions of people and livestock. Anthelmintic drugsareavailablebutresistanceisaprobleminlivestockparasites,andisaloomingthreatforhumanhelminths.Testing the efficacy of available anthelmintic drugs and development of new drugs is hindered by the lack of objective high- throughputscreeningmethods.Currently,drugeffectisassessedbyobservingmotilityordevelopmentofparasitesusing laborious,subjective,low-throughputmethods. Methodology/PrincipalFindings:Herewedescribeanovelapplicationforareal-timecellmonitoringdevice(xCELLigence) thatcansimplyandobjectivelyassessanthelminticeffectsbymeasuringparasitemotilityinrealtimeinafullyautomated high-throughputfashion.WequantitativelyassessedmotilityanddeterminedrealtimeIC50valuesofdifferentanthelmintic drugs against several developmental stages of major helminth pathogens of humans and livestock, including larval HaemonchuscontortusandStrongyloidesratti,andadulthookwormsandbloodflukes.Theassayenabledquantificationof theonsetofegghatchinginrealtime,andtheimpactofdrugsonhatchrate,aswellasdiscriminatingbetweentheeffects ofdrugsonmotilityofdrug-susceptibleand–resistantisolatesofH.contortus. Conclusions/Significance:Ourfindingsindicatethatthistechniquewillbesuitablefordiscoveryanddevelopmentofnew anthelmintic drugs as well as for detection of phenotypic resistance to existing drugs for the majority of helminths and otherpathogenswheremotilityisameasureofpathogenviability.Themethodisalsoamenabletouseforotherpurposes wheremotilityisassessed,suchasgenesilencingorantibody-mediatedkilling. Citation:SmoutMJ,KotzeAC,McCarthyJS,LoukasA(2010)ANovelHighThr