title: Differential identification of Chlamydophila abortus live vaccine strain 1B and C. abortus field isolates by PCR-RFLP.
abstract: Comparative genomic analysis of a wild-type strain of the ovine pathogen Chlamydophila abortus and its nitrosoguanidine-induced, temperature-sensitive and virulence-attenuated live vaccine derivative identified point mutations unique to the mutant (Burall et al. [1]). Here, we evaluate the capacity of some of these mutations to either create or eliminate restriction sites using the wild-type strain C. abortus S26/3 as a reference. Three of eight genomic sites with confirmed point mutations (CAB153, CAB636 and CAB648) were retained for analysis as each resulted in the loss of a restriction site in the genome sequence of the vaccine strain. PCR-restriction fragment length polymorphism analysis using restriction enzymes chosen to specifically target the three genomic sites was then applied to a large number of C. abortus field isolates and reference strains. Our results indicate that the three mutations are uniquely present in the vaccine strain, and as such provide easy-to-use markers for the differential identification of the vaccine strain and wild-type isolates.
authors.affiliation: Unité Zoonoses Bactériennes, Agence Française de Sécurité Sanitaire des Aliments (Lerpaz), 23 Avenue du Général de Gaulle, 94706 Maisons-Alfort Cedex, France. k.laroucau@afssa.fr
authors.forename: Karine
authors.initials: K
authors.lastname: Laroucau
authors.forename: Fabien
authors.initials: F
authors.lastname: Vorimore
authors.forename: Konrad
authors.initials: K
authors.lastname: Sachse
authors.forename: Evangelia
authors.initials: E
authors.lastname: Vretou
authors.forename: Victoria I
authors.initials: VI
authors.lastname: Siarkou
authors.forename: Hermann
authors.initials: H
authors.lastname: Willems
authors.forename: Simone
authors.initials: S
authors.lastname: Magnino
authors.forename: Annie
authors.initials: A
authors.lastname: Rodolakis
authors.forename: Patrik M
authors.initials: PM
authors.lastname: Bavoil
mesh_terms: D015373:Bacterial Typing Techniques; D001428:Bacterial Vaccines / Q000382:microbiology; D023501:Chlamydophila / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D055028:Comparative Genomic Hybridization; D004269:DNA, Bacterial / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D001428:Bacterial Vaccines; D004269:DNA, Bacterial