A Low-Cost Microfluidic Chip for Rapid Genotyping of Malaria-Transmitting Mosquitoes 英文参考文献.docVIP

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A Low-Cost Microfluidic Chip for Rapid Genotyping of Malaria-Transmitting Mosquitoes 英文参考文献.doc

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A Low-Cost Microfluidic Chip for Rapid Genotyping of Malaria-Transmitting Mosquitoes 英文参考文献

ALow-CostMicrofluidicChipforRapidGenotypingof Malaria-TransmittingMosquitoes ChangchunLiu1,MichaelG.Mauk1,RobertHart1,MariangelaBonizzoni2,GuiyunYan2,HaimH.Bau1* 1DepartmentofMechanicalEngineeringandAppliedMechanics,UniversityofPennsylvania,Philadelphia,Pennsylvania,UnitedStatesofAmerica,2CollegeofHealth Sciences,UniversityofCaliforniaIrvine,Irvine,California,UnitedStatesofAmerica Abstract Background:Vectorcontrolisoneofthemosteffectivemeasurestopreventthetransmissionofmalaria,adiseasethat causesover600,000deathsannually.Around30–40Anopheles mosquitospeciesarenaturalvectorsofmalariaparasites. Some of these species cannot be morphologically distinguished, but have behavioral and ecological differences. EmblematicofthisistheAnophelesgambiaespeciescomplex.Thecorrectidentificationofvectorspeciesisfundamentalto thedevelopmentofcontrolstrategiesandepidemiologicalstudiesofdiseasetransmission. Methodology/Principal Findings: An inexpensive, disposable, field-deployable, sample-to-answer, microfluidic chip was designed, constructed, and tested for rapid molecular identification of Anopheles gambiae and Anopheles arabiensis . The chipcontainsthreeisothermalamplificationreactors.OnetestreactoroperateswithspecificprimerstoamplifyAnopheles gambiae DNA, another with specific primers for Anopheles arabiensis DNA, and the third serves as a negative control. A mosquito leg was crushed on an isolation membrane. Two discs, laden with mosquito tissue, were punched out of the membraneandinsertedintothetwotestchambers.Theisolated,disc-boundDNAservedasatemplateintheamplification processes.Theamplificationproductsweredetectedwithintercalatingfluorescentdyethatwasexcitedwithabluelight- emittingdiode.Theemittedlightwasobservedbyeyeandrecordedwithacell-phonecamera.Whenthetargetconsisted ofAnophelesgambiae,thereactorcontainingprimersspecifictoAn.gambiaelitupwhiletheothertworeactorsremained dark.WhenthetargetconsistedofAnophelesarabiensis,thereactorcontainingprimersspecifictoAn.arabiensis