Drug-Resistant Genotypes and Multi-Clonality in Plasmodium falciparum Analysed by Direct Genome Sequencing from Peripheral Blood of Malaria Patients 英文参考文献.docVIP

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Drug-Resistant Genotypes and Multi-Clonality in Plasmodium falciparum Analysed by Direct Genome Sequencing from Peripheral Blood of Malaria Patients 英文参考文献.doc

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Drug-Resistant Genotypes and Multi-Clonality in Plasmodium falciparum Analysed by Direct Genome Sequencing from Peripheral Blood of Malaria Patients 英文参考文献

Drug-ResistantGenotypesandMulti-Clonalityin PlasmodiumfalciparumAnalysedbyDirectGenome SequencingfromPeripheralBloodofMalariaPatients TimothyRobinson1.,SusanaG.Campino2.,SarahAuburn2,3,SamuelA.Assefa2,SpencerD.Polley4, MagnusManske2,BronwynMacInnis1,2,KirkA.Rockett1,2,GarethL.Maslen2,MandySanders2, MichaelA.Quail2,PeterL.Chiodini4,5,DominicP.Kwiatkowski1,2,TaaneG.Clark5,ColinJ.Sutherland4,5* 1WellcomeTrustCentreforHumanGenetics,UniversityofOxford,Oxford,UnitedKingdom,2WellcomeTrustSangerInstitute,Hinxton,UnitedKingdom,3GlobalHealth Division,MenziesSchoolofHealthResearch,CharlesDarwinUniversity,Darwin,Australia,4DepartmentofClinicalParasitology,HospitalforTropicalDiseases,London, United Kingdom, 5Faculties of Infectious and Tropical Diseases and Epidemiology and Population Health, London School of Hygiene Tropical Medicine, London, UnitedKingdom Abstract Naturallyacquiredblood-stageinfectionsofthemalariaparasitePlasmodiumfalciparumtypicallyharbourmultiplehaploid clones. The apparent number of clones observed in any single infection depends on the diversity of the polymorphic markersusedfortheanalysis,andtherelativeabundanceofrareclones,whichfrequentlyfailtobedetectedamongPCR productsderivedfromnumericallydominantclones.However,minorityclonesareofclinicalinterestastheymayharbour genes conferring drug resistance, leading to enhanced survival after treatment and the possibility of subsequent therapeutic failure. We deployed new generation sequencing to derive genome data for five non-propagated parasite isolatestakendirectlyfrom4differentpatientstreatedforclinicalmalariainaUKhospital.Analysisofdepthofcoverageand length of sequence intervals between paired reads identified both previously described and novel gene deletions and amplifications.Full-lengthsequencedatawasextractedfor6lociconsideredtobeunderselectionbyantimalarialdrugs, and both known and previously unknown amino acid substitutions were identified. Full mitochondrial genomes were extractedfromthesequencing