肺炎链球菌的耐药基因及其分子生物学检测

201008238MedicalInformation.Aug.2010.Vol.23.No.8。TQ。hPlk1-PBDTQPXNPPG2-PXP。。Watanabe[7]PPG2-Pyrogallin6PBD。3.3hPlk1-PBDPXN。。PXPPPGPBD。PPG2-。。。-。hPlk1-PBD。：[1]BanerjeeS,KasebAO,WangZ,KongD,MohammadM,PadhyeS,SarkarFH,MohammadRM.CancerRes.2009,69,5575-83.[2]SalomiJ,NairC,JayawardhananK,VargheseD,PanikkarR.,CancerLett,1992,63:41-46.[3]SalomiJ,NairC,PanikkarR.NutrandCancer,199116,67-72.[4]ReindlW.,YuanJ.,Kr?merA.,StrebhardtK.BergT.ChemBiol,2008,15,459-66.[5]MundtKE,GolsteynRM,LaneHA,NiggEA.BiochemBiophysResCommun.1997,239,377-385.[6]García-AlvarezB.,deCárcerG.,Iba?ezS,Bragado-NilssonE.MontoyaG.ProcNatlAcadSciUSA,2007,104,3107-3112.[7]WatanabeN,SekineT,TakagiM,IwasakiJ,ImamotoN,KawasakiH,OsadaH.JBiolChem,2009284,2344-2353.[8]ReindlW.,YuanJ.,Kr?merA.,StrebhardtK.,BergT.Chembiochem,2009,10,1145-1148.[9]LeeKS.,IdleJR.ChemBiol,2008,15,459-466[10]ChenzhongLiao,Jung-EunPark,JeongK.Bang,MarcC.Nicklaus,KyungS.LeeACSMedicinalChemistryLetters,2010,9[11]IvankovicS,StojkovicR,JukicM,MilosM,MilosM,JurinM.ExpOncol,2006,28:220-224.[12]WorthenDR,GhoshehOA,CrooksPA.AnticancerRes,199818,1527-1532./2DithymoquinonePyrogallinDithymoquinone5Pyrogallin63p-78910(StreptococcuspneumoniaeSPN)。1990,SPN,。SPN、、SPN[1]SPNpbperm。SPNpbperm。1SPN(pen-icillinresistantStreptococcuspneumoniaePRSP)SPN10.0%～50.0%[12]。:[beta]-lactam,[3]pe-nicillin-bindingproteins,PBPs。PBP[4～6]。PBP。。S.pneumoniaePBPsPBP1A、PBP2BPBP2X。PBP2X（,636400）：β－，，，。β－，pbpβ－，MLS，MermBmefA。，。：；；pbp；erm；2010-06-103055201008238MedicalInformation.Aug.2010.Vol.23.No.8。PBPPBP2BPBP2X。PBP1APBP2BPBP2X[6～8]。SPNPBP2BTER(Transpeptidase-encodingregion)PSSP(penicillin-susceptibleS.pneumoniae)。PBP2B。SPNPBP2BABAB。SPNPCR。PCRpbp1a,pbp2b,pbp2xDNA[6]。SPNPCRP1P2SPNpbp2bP2。:R1)5'-GCCTTTTCTAGGCCAATGCCGATTAC-3'R25'-GCCTACGATTCATTCCCGATT-3',R35'-AAATTGGCATATGGATCTTTTCCT-3'R45'-GTTTTAACTAACAATTTAGAATCC-3'P55'-CTGACCATTGATTTGGTTTCCAA-3'P65'-TTTGCAATAGTTGCTACATACTG-3'。。n-PCR[9]。P15'-CTGACCATTGATTTGGTTTCCAA-3'P25'-TTTGCAATAGTTGCTACATACTG-3'P35'-CCTGATTCCTTGGGAACGGT-3'P45'-CCCAAGCCATATTCGCCAAA-3'。(penicillin-nonsusceptibleS.pneumoniae,PNSP)。R6pbp2bPNSP13Glu1393Gly,Ile1673Leu,Asn2283Tyr,Thr2323Lys,Gln2333Leu,Gln2443Glu,Thr2523Ala,Leu2613Ile,Ser2793Thr,Glu2823Gly,Ser2863Ala,Thr2963Ala,andAsn3443Asp。(blockB,positions367to482)。MultiplexPCR。(P1:149-F1,5'-TTATGCAGTTGCTTTGAAT-3'P2155-F2,5'-AACCCWAARACAGGTGCT-3'nucleotides447to465and463to480inR6,respectively(322-R,5'-GTTATCAAACTGCCCAAAGGC-3'nucleotides963to984inR6)。149-F1155-F2PNSP[10]。SPN[beta]-lactam。23SPNTEMSHV[beta]-lactam[11]。21TEMDNA。TEM。SPN。2SPNSPN,1997~20008.8%~22.5%,42.5%~76.8%[1213]。erm。Granizor=0.942[14]。SPN①ermBMLSB。cMLSiMLS。ermermBermTR。②mefAmefA3[15]。141516BM。③23SrRNAL4。。M[16]。HobanermBmefAP15'-GAAAAGGTACTAAACCAAATA-3',P25'-AGTAACCGGTACTTAAATTGTTTAC-3',616bpP15'-ACTATCATTAATCACTAGTGC-3',P25'-TTCTTCTGGTACTAAAAGTGC-3'346bp。ermA/BPCRP15'-GA(A/G)ATIGGIIIIGGIAA(A/G)GGICA-3',P25'-AA(C/T)TG(A/G)TT(C/T)TTIGT(A/G)AA-3',I533pb[17]。ermmefpbp。。tetMribosomalprotectionproteins,RPPs[1819]。：[1].、2000～2001[J].200248461~466.[2]．1999～20004、[J]．20011(3)142~146．[3]LivermoreDM.[beta]-lactamasesinlaboratoryandclinicalresistance[J].ClinMicrobiolRev．1995,8557~58.[4]duPlessisMBingenEKlugmanKP．Analysisofpenicillin-bindingpro_teingenesofclinicalisolatesofStreptococcuspneumoniaewithreducedsusceptibilitytoamoxicillin[J]．AntimicrobAgentsChemother200246(8)2349~2357．[5]NagaiKDaviesTAJacobsMR．eta1．Effectsofaminoacidalterationsinpenicillin-bindingproteins(PBPs)1a2band2xonPBPaffinitiesofpenicillinamoxicillincefditorencefuroxime．cefprozilandcefaclorin18clinicalisolatesofpenicillinsusceptibleintermediateandresistantpneumococci[J]．AntimicrobAgentsChemother200246(5)1273~1280．[6]NicholKA,ZhanelGG,HobanDJ.Penicillin-bindingprotein1A,2B,and2XalterationsinCanadianisoletsofpenicillin-resistantStreptococcuspneumoniae[J].AntimicrobAgentsChemother2002,46:3261~3264.[7]CharpentierE,TuomanenE.MechanismsofantibioticresistanceandtoleranceinStreptococcuspneumoniae[R].MicrobesInfect2000,2:1855~1864.[8]HakenbeckR.TransformationinStreptococcuspneumoniae:mosaicgenesandtheregulationofcompetence[J]．ResMicrobiol2000,151:453~456.[9].pbp2b[J].,200528855~856.[10]JinYangBaek,KwanSooKo,WonSupOh,2Sook-InJung,YeonSookKim,3056201008238MedicalInformation.Aug.2010.Vol.23.No.8Hyun-HaChang,HyuckLee,ShinWooKim,KyongRanPeck,NamYongLee,andJae-HoonSong.UniqueVariationsofpbp2bSequencesinPenicillin-NonsusceptibleStreptococcuspneumoniaeIsolatesfromKorea[J]．JOURNALOFCLINICALMICROBIOLOGY.2004,42:1746~1750.[11].ESBLs[J].200315142~143.[12]WeinsteinAJ.[J]．2001818~16.[13].[J]．200038424~427.[14]GranizoJJ,AguilarL,CasalJ,etal.Streptococcuspneumoniaeresistancetoerythromycinandpenicillininrelationtomacrolideandbeta-lactamconsumptioninSpian(1979~1997)[J].Antimicrob.Chemother,2000,46:767~773.[15]RobertsMC,SutcliffeJ,ourvalinP,etal.Nomenclatureformacrolideandmacrolide-Lincosamide-StreptogtraminBresistancedeterminants[J]．Antimicrob.AgentsChemother,1999,43:2826~2830.[16].[J]．200543329~332.[17]HobanDJ,WierzbowskiAk,NicholK,etal.Macrolide-resistantStrept