Sequence of Closely Related Plasmids Encoding blaNDM-1 in Two Unrelated Klebsiella pneumoniae Isolates in Singapore 英文参考文献.docVIP

SequenceofCloselyRelatedPlasmidsEncodingblaNDM-1 inTwoUnrelatedKlebsiellapneumoniaeIsolatesin Singapore Ying-TsongChen1,2,3,Ann-ChiLin1,L.KristopherSiu4,5*,TseHsienKoh6* 1InstituteofMolecularandGenomicMedicine,NationalHealthResearchInstitutes,Zhunan,Taiwan,2InstituteofGenomicsandBioinformatics,NationalChungHsing University,Taichung,Taiwan,3Biotechnology Center,National ChungHsing University, Taichung, Taiwan, 4NationalInstitute of Infectious Diseases and Vaccinology, NationalHealthResearchInstitutes,Miaoli,Taiwan,5GraduateInstituteofBasicMedicalScience,ChinaMedicalUniversity,Taichung,Taiwan,6DepartmentofPathology, SingaporeGeneralHospital,Singapore,Singapore Abstract Background:SpreadoftheblaNDM-1 genethatencodestheNewDelhimetallo-b-lactamase(NDM-1)inEnterobacteriaceae isamajorglobalhealthproblem.PlasmidscarryingblaNDM-1 fromtwodifferentmulti-drugresistantKlebsiellapneumonia isolatescollectedinSingaporewerecompletelysequencedandcomparedtoknownplasmidscarryingblaNDM-1 . Methodology/PrincipalFindings:Thetwoplasmids,pTR3andpTR4,weretransferredtoEscherichiacolirecipientstrainJ53 and completely sequenced by a shotgun approach using 3-kb paired-end libraries on 454. Although the K. pneumoniae strainswereunrelatedbymoleculartypingusingPFGEandMLST,completesequencingrevealedthatpTR3andpTR4are identical.TheplasmidsequenceissimilartotheE.coliNDM-1-encodingplasmidp271A,whichwasisolatedinAustraliafrom apatientreturningfromBangladesh.TheimmediateregionsoftheblaNDM-1 geneinpTR3/4areidenticaltothatofp271A, but the backbone of our plasmid is much more similar to another IncN2 plasmid reported recently, pJIE137, which containedanadditional5.2-kbCUP(conservedupstreamrepeat)regulonregionincomparisontop271A.A257-bpelement boundedbyimperfect39-bpinvertedrepeats(IR)andanincompleteversionofthiselementflankingthe3.6-kbNDM-1- encodingregionwereidentifiedintheseplasmidsandarelikelytobethevestigesofanunknownIS. Conclusions:Althoughthehostsarenotepidemiologicallylinked,wefoundthatt