国内部分地区多杀性巴氏杆菌荚膜血清型和基因型的研究
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摘要
为了解国内部分地区不同动物多杀性巴氏杆菌(P.multicida)流行情况,本研究以36株P.multicida为研究对象,通过多重荚膜PCR、多位点序列分型(MLST)和脂多糖多重PCR(LPS-m PCR)对其荚膜型和基因型进行鉴定。采用多重荚膜PCR方法将36株细菌的荚膜血清型分为A、B和D 3种,其中禽源P.multicida主要以A为主;以LPS-m PCR方法将P.multicida分为L1、L2、L3和L6 4种LPS基因型,禽源P.multicida主要为L1型;通过MLST技术将P.multicida分为ST-129、ST-8、ST-58、ST-5、ST-13、ST-50和ST-122 7种ST型,禽源P.multicida主要为ST-129型。本研究为P.multicida的流行病学监测和基因多样性提供了技术支撑。
To investigate the epidemic status of the Pasteurella multocida isolates from different hosts in some areas of China,thirty-six P.multocida isolates were isolated from the clinical samples collected from 6 provinces,and subject to identifications of the capsule types and genotypes by multiplex capsular PCR,MLST and LPS-m PCR.The isolates were found to be members of capsule serotypes A,B,and D based on capsular serotyping using multiplex capsular PCR,of which the avian original isolates were mainly classified into capsule type A.Four different LPS genotypes,included L1,L2,L3,and L6,were identified by LPS-m PCR,and avian original isolates were mainly in L1.Seven different sequence types(STs) including ST-129,ST-8,ST-58,ST-5,ST-13,ST-50,and ST-122,were identified by MLST,and avian original isolates were mainly for ST-129.These results demonstrated MLST and LPS-m PCR approach are useful for epidemiological surveillance and determination of the genetic diversity of P.multocida prevalance.
To investigate the epidemic status of the Pasteurella multocida isolates from different hosts in some areas of China,thirty-six P.multocida isolates were isolated from the clinical samples collected from 6 provinces,and subject to identifications of the capsule types and genotypes by multiplex capsular PCR,MLST and LPS-m PCR.The isolates were found to be members of capsule serotypes A,B,and D based on capsular serotyping using multiplex capsular PCR,of which the avian original isolates were mainly classified into capsule type A.Four different LPS genotypes,included L1,L2,L3,and L6,were identified by LPS-m PCR,and avian original isolates were mainly in L1.Seven different sequence types(STs) including ST-129,ST-8,ST-58,ST-5,ST-13,ST-50,and ST-122,were identified by MLST,and avian original isolates were mainly for ST-129.These results demonstrated MLST and LPS-m PCR approach are useful for epidemiological surveillance and determination of the genetic diversity of P.multocida prevalance.
引文
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[2]Blackall P J,Miflin J K.Identification and typing of Pasturella multocida:a review[J].Avian Pathol,2000,29(4):271-287.
[3]Townsend K M,Dawkins H J,Papadimitriou J M.REP-PCR analysis of Pasteurella multocida isolates that cause haemorrhagicsepticaemia[J].Res Vet Sci,1997,63(2):151-155.
[4]Loubinoux J,Lozniewski A,Lion C,et al.Value of enterobacterial repetitive intergenic consensus PCR for study of Pasteurella multocida strains isolated from mouths of dogs[J].J Clin Microbiol,1999,37(8):2488-2492.
[5]Amonsin A,Wellehan J F,Li L L,et al.DNA fingerprinting of Pasteurella multocida recovered from avian sources[J].J ClinMicrobiol,2002,40(8):3025-3031.
[6]Davies R L,MacCorquodale R,Reilly S.Characterization of bovine strains of Pasteurella multocida and comparison with isolates of avian,ovine and porcine origin[J].Vet Microbiol,2004,99(2):145-158.
[7]Subaaharan S,Blackall L L,Blackall P J.Development of a multilocus sequence typing scheme for avian isolates of Pasteurella multocida[J].Vet Microbiol,2010,141(3-4):354-361.
[8]Davies R L,MacCorquodale R,Caffrey B.Diversity of avian Pasteurella multocida strains based on capsular PCR typing and variation of the OmpA and OmpH outer membrane proteins[J].Vet Microbiol,2003,91:169-182.
[9]Wang Yan-hong,Zhu Jie,Lu Cheng-ping,et al.Evidence of circulation of an epidemic strain of Pasteurella multocida in Jiangsu,China by multi-locus sequence typing(MLST)[J].Infect Genet Evol,2013,20:34-38.
[10]Harper M,St Michael F.Structure and biosynthetic locus of the lipopolysaccharide produced by Pasteurella multocida serovars 8and 13 and the identification of a novel phospho-glycero moiety[J].Glycobiology,2013,23(3):286-294.
[11]Harper M,John M,Turnim C,et al.Development of a rapid multiplex PCR assay to genotype Pasteurella multocida strains by use of the lipopolysaccharide outer core biosynthesis locus[J].J Clin Microbiol,2015,53(2):477-485.
[12]Townsend K M.Genetic Organization of Pasteurella multocida cap loci and development of a multiplex capsular PCR typing system[J].J Clin Microbiol,2001,39(3):924-929.
[13]Petersen A,Bisgaard M.MLST typing of Pasteurella multocida associated with haemorrhagicsepticaemia and development of a real-time PCR specific for haemorrhagicsepticaemia associated isolates[J].Vet Microbiol,2014,170:335-341.