Differential identification of Chlamydophila abortus live vaccine strain 1B and C. abortus field isolates by PCR-RFLP |
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| Authors: | Karine Laroucau Fabien Vorimore Konrad Sachse Evangelia Vretou Victoria I. Siarkou Hermann Willems Simone Magnino Annie Rodolakis Patrik M. Bavoil |
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| Affiliation: | 1. Unité Zoonoses Bactériennes, Agence Française de Sécurité Sanitaire des Aliments (Lerpaz), 23 Avenue du Général de Gaulle, 94706 Maisons-Alfort Cedex, France;2. Friedrich-Loeffler-Institut (Federal Research Institute for Animal Health), Institute of Molecular Pathogenesis, Naumburger Str. 96a, 07743 Jena, Germany;3. Laboratory of Biotechnology, Hellenic Pasteur Institute, Vassilissis Sofias Avenue 127, 11521 Athens, Greece;4. Laboratory of Microbiology and Infectious Diseases, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, University Campus, GR-54124 Thessaloniki, Greece;5. Institute for Hygiene and Infectious Diseases of Animals, University of Giessen, Germany;6. Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna “Bruno Ubertini”, National Reference Laboratory for Animal Chlamydioses, Sezione Diagnostica di Pavia, Strada Campeggi 61, 27100 Pavia, Italy;g INRA, UR1282 Infectiologie Animale et Santé Publique, 37380 Nouzilly, France;h University of Maryland, Department of Microbial Pathogenesis, Baltimore, MD 21201, USA |
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| 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. |
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| Keywords: | Chlamydophila abortus Live vaccine Identification |
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