猪链球菌分子生物学分型方法研究进展

,2010,31(10):8387ProgressinVeterinaryMedicine*林荣高,黎满香*,蒋伟,陈滔(,410128):猪链球菌是猪重要的致病菌,其种类繁多,鉴定困难传统分型方法包括兰氏分群法,荚膜抗原分型,溶血性分型及生化分型法等分子生物学技术的发展给猪链球菌的分型带来新的思路论文对猪链球菌分子生物学分型方法进行介绍,包括多位点序列分型技术脉冲场凝胶电泳扩增片段长度多态性随机扩增的多态性DNA限制性片段长度多态性核糖体分型,PCR限制性片段长度多态性和测序:猪链球菌;分子生物学;分型:S852.611:A:10075038(2010)10008305(Streptococcussuis),,,[1],[23]1968,2008400[4],[5]19982005,,,(lancefield),RST,,[6],,,[7],35,1~341/2,[8],2,,2(SS2)[9],,,,,,1(multilocussequencetyping,MLST)PCRKingSJ[10]2944,92(ST),ST1,6141,ST1,ST27,ST87,PrincivalliMS[11]2003-2007ST1,MRP,EF,suilysin,ST7,ST1(19982005),114106ST7,[1213],5SS23ST7,2ST1,ST7TST1[14]DangH[15]16,MLST,,16,MLST,,,,*:20100330:湖南省科技计划项目(06FJ4101):林荣高(1984-),男,广西梧州人,硕士研究生,主要从事动物传染病研究*2(pulsedfieldgelelectrophoresis,PFGE),,,DNAVelaAI[16]7430260,45%,(pulsotypes),92724106,MaroisC[17],58%,,PFGEBerthelotHeraultF[18]123PFGE,98%74,60%ABC3,B,PFGE,PFGE,,,MLST,PFGE,,,[19]3MogollondJD[20](RFLP),DNA,HaeHind,,DNAHae23,9111216DNAHaeHind916110,DNA,DNAAmassSF[21]RFLP,35,RFLP:DNA,DNADNA,,RFLP,AFLPPFGE,4(amplifiedfragmentlengthpolymorphism,AFLP)RFLP,RehmT[22]116AFLPABCD4A1A2C1C2A1cps1mrp+epf+(epf*)sly+cps2+;mrp*cps9+A2;C1C2cps2cps7epf-sly-AFLP,BaumsCG[23]83S.suisAFLPmrpepfslycps2cps2+,sly-epf-SS2,AFLP45%,AFLPAFLP,,,DNAPCR5DNADNA(randomamplifiedpolymorphicDNA,RAPD)DNA,DNAChatellierS[24]88SS2,23DNA,,SLYMRPEF,SLY+MRP+EP+PAPD,SLY-MRP-EP-,,RAPD,,RAPD,,,,6StaatsJJ[25](ribotyping)8420103110(208),DNASouthernblot,16S23SrRNAcDNA,SS2SmitsHE[26]1066bp16sRNA42DNA,SS2SS1,[27]DNA,,Southern,,,7PCRPCR(PCRRFLP)PCRDNA,RFLPMaroisC[28]138,PCR16SrRNA23SrRNA,16S~23SrRNA(intergenicspacerregion,ISR),ISRRFLP,95%,,SS223SrRNA(3kb),,ISR16SrRNA,16S~23SrRNAISR16SrRNADNA,PCRRFLP,DNA8(sequencing)16SDNARasmussenSR[29]16SDNA,92026,2527224122ChatellierC[30]3516SRNA,93.49%~100%,16SRNA16SRNA32,33,34,16SRNA,,,60kuchaperonins(Cpn60)BrousseauR[31]35Cpn60,Cpn6016SRNA,Cpn6016SRNA,KutzR[32]19SS2(glutamatedehydrogenaseenzymes,GDH),GDH1820bp,1344,,214886895913988Tyr72toAspThr296toAlaAla299toSerGlu305toLysGlu330toLys,Ala299SerGlu305LysGlu330Lys,GDH9,,,,,,,PAPDAFLPRFLP,PAPD,,AFLP,,PCRRFLP,,PCRRFLP,,MLSTPFGE,,,,ZuckerkandlEPaulingL![33],,:[1].[M].:,2005:211.[2]MaraC,PunaroD,SeguraM.Streptococcussuisserotype2,animportantswineandhumanpathogen,inducesstrongsys85:temicandcerebralinflammatoryresponsesinamousemodelofinfection[J].Immunology,2007,179:18421854.[3]MaY,FengYJ,LiuD,etal.Avianinfluenzavirus,Streptococcussuisserotype2,severeacuterespiratorysyndromecoronavirusandbeyond:molecularepidemiology,ecologyandthesituationinChina[J].PhilTransRSocB,2009,364:27252737.[4]WertheimHF,NghiaHD,TaylorW,etal.Streptococcussuis:anemerginghumanpathogen[J].ClinInfectDis,2009,48:617625.[5]SmithTC,CapuanoAW,BoeseB,etal.OccupationalexposuretoStreptococcussuisamongUSswineworkers[J].EmergInfectDis,2008,14:19251927.[6],.RAPD[J].,2002,23(2):2932.[7],,.[J].,2007,41(3):2931.[8]PanXZ,GeJC,LiM,etal.TheorphanresponseregulatorcovR:agloballynegativemodulatorofvirulenceinStreptococcussuisserotype2[J].Bacteriol,2009,191:26012612.[9]GottschalkM,XuJ,CalzasC,etal.Streptococcussuis:anewemergingoranoldneglectedzoonoticpathogen[J].FutureMicrobiol,2010,5(3):371391.[10]KingSJ,LeighJA,HeathPJ,etal.DevelopmentofamultilocussequencetypingschemeforthepigpathogenStreptococcussuis:Identificationofvirulentclonesandpotentialcapsularserotypeexchange[J].ClinMicrobiol,2002,40(10):36713680.[11]PrincivalliMS,PalmieriC,MagiG.GeneticdiversityofStreptococcussuisclinicalisolatesfrompigsandhumansinItaly(20032007)[J].EuroSurveill,2009,14(33):pii:19310.[12]YeCY,BaiXM,ZhangJ,etal.SpreadofStreptococcussuissequencetype7inChina[J].EmergInfectDis,2008,14(5):787791.[13],,,.2005MLST[J].,2008,28(8):14381441,1445.[14]YeC,ZhengH,ZhangJ,etal.Clinical,experimental,andgenomicdifferencesbetweenintermediatelypathogenic,highlypathogenic,andepidemicStreptococcussuis[J].InfectDis,2009,199(1):97107.[15]DangH,NghiaT,HoaNT,etal.HumancaseofStreptococcussuisserotype16infection[J].EmergInfectDis,2008,14(1):155157.[16]VelaAI,GoyacheJ,TarradasC.AnalysisofgeneticdiversityofStreptococcussuisclinicalisolatesfrompigsinspainbypulsedfieldgelelectrophoresis[J].ClinMicrobiol,2003,41(6):24982502.[17]MaroisC,DevendecLL,GottschalkM,etal.DetectionandmoleculartypingofStreptococcussuisintonsilsfromlivepigsinFrance[J].CanJVetRes,2007,71(1):1422.[18]BerthelotHeraultF,MaroisC,GottschalkM,etal.GeneticdiversityofStreptococcussuisstrainsisolatedfrompigsandhumansasrevealedbypulsedfieldgelelectrophoresis[J].ClinMicrobiol,2002,40(2):615619.[19],,,.[J].,2008,29(1):7073.[20]MogollonJD,PijoanC,MurtaughMP.CharacterizationofprototypeandclinicallydefinedstrainsofStreptococcussuisbygenomicfingerprinting[J].ClinMicrobiol,1990,28:24622466.[21]AmassSF,SanmiguelsP,ClarkLK.DemonstrationofverticaltransmissionofStreptococcussuisinswinebygenomicfingerprinting[J].ClinMicrobiol,1997,35(6):15951959.[22]RehmT,BaumsCG,StrommengerB,etal.AmplifiedfragmentlengthpolymorphismofStreptococcussuisstrainscorrelateswiththeirprofileofvirulenceassociatedgenesandclinicalbackground[J].MedicalMicrobiol,2007,56:102109.[2