Vaccine. 2010 Aug 9;28(35):5653-6. Epub 2010 Jun 29.

Differential identification of Chlamydophila abortus live vaccine strain 1B and C. abortus field isolates by PCR-RFLP.

Laroucau K, Vorimore F, Sachse K, Vretou E, Siarkou VI, Willems H, Magnino S, Rodolakis A, Bavoil PM.

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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

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.

PMID:: 20600479; [PubMed - indexed for MEDLINE]

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