title: Genetic variability of Chlamydophila abortus strains assessed by PCR-RFLP analysis of polymorphic membrane protein-encoding genes.
abstract: This study used PCR-RFLP to investigate the genetic variability of pmp-encoding genes from fifty-two Chlamydophila abortus (C. abortus) strains originating from abortion cases from various geographical regions and host species. Six primer pairs were used to PCR-amplify DNA fragments encoding eighteen pmps. PCR products were digested using four restriction endonucleases and Bayesian methodologies were used to compare RFLP profiles and assign strains to a RFLP genotype. Strains could be assigned to 2 genotypes in the region encoding pmp18D, 3 genotypes in the regions encoding pmp1A-pmp2B, pmp3E-pmp6H and pmp11G-pmp15G, 4 genotypes in the region encoding pmp7G-pmp10G and 5 genotypes in the region encoding pmp16G-pmp17G. In all regions, the majority of strains (88.4-96.1%) had the same genotype as the reference strain S26/3. No correlation could be made between genotype, host species or geographical origin except for the two variant Greek strains, LLG and POS, which formed a discrete genotype in all pmp-encoding regions except pmp18D. Relative rates of evolution calculated for each pmp-encoding gene locus suggest that differing selective pressures and functional constraints may exist on C. abortus polymorphic membrane proteins. These findings suggest that although intraspecies heterogeneity of pmp-encoding genes in C. abortus is low, the sequence heterogeneity should be an important consideration when using pmps as the basis for novel diagnostics or vaccine development.
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Bush Loan, Edinburgh, Midlothian EH26 0PZ, UK. michelle.sait@moredun.ac.uk
authors.forename: Michelle
authors.initials: M
authors.lastname: Sait
authors.forename: Ewan M
authors.initials: EM
authors.lastname: Clark
authors.forename: Nicholas
authors.initials: N
authors.lastname: Wheelhouse
authors.forename: Lucy
authors.initials: L
authors.lastname: Spalding
authors.forename: Morag
authors.initials: M
authors.lastname: Livingstone
authors.forename: Konrad
authors.initials: K
authors.lastname: Sachse
authors.forename: Bryan K
authors.initials: BK
authors.lastname: Markey
authors.forename: Simone
authors.initials: S
authors.lastname: Magnino
authors.forename: Victoria I
authors.initials: VI
authors.lastname: Siarkou
authors.forename: Evangelia
authors.initials: E
authors.lastname: Vretou
authors.forename: María R
authors.initials: MR
authors.lastname: Caro
authors.forename: Raja
authors.initials: R
authors.lastname: Yaga
authors.forename: F Alex
authors.initials: FA
authors.lastname: Lainson
authors.forename: David G E
authors.initials: DG
authors.lastname: Smith
authors.forename: Frank
authors.initials: F
authors.lastname: Wright
authors.forename: David
authors.initials: D
authors.lastname: Longbottom
mesh_terms: D000034:Abortion, Veterinary / Q000382:microbiology*; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D001499:Bayes Theorem; D023501:Chlamydophila / Q000145:classification / Q000235:genetics*; D023521:Chlamydophila Infections / Q000662:veterinary*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D005843:Geography; D058751:Livestock / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D011247:Pregnancy
chemical_list: D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers