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Clinical and Diagnostic Laboratory Immunology, Sep 1997, 620-623, Vol 4, No. 5
Copyright © 1997 by the American Society for Microbiology. All rights reserved.

Development of an in vitro cloning method for Cowdria ruminantium

JM Perez, D Martinez, A Debus, C Sheikboudou and A Bensaid
Departement Elevage et Medecine Veterinaire, Centre International en Recherche Agronomique pour le Developpement, Pointe a Pitre, Guadeloupe, French West Indies.

Cowdria ruminantium is a tick-borne rickettsia which causes severe disease in ruminants. All studies with C. ruminantium reported so far were carried out with stocks consisting of infective blood collected from reacting animals or from the same stocks propagated in vitro. Cloned isolates are needed to conduct studies on immune response of the host, on genetic diversity of the parasite, and on mechanisms of attenuation and the development of vaccines. A method of cloning based on the particular chlamydia life cycle of Cowdria was developed. Instead of cloning extracellular elementary bodies, it appeared more convenient to clone endothelial cells infected by one morula resulting from the infection of the cell by one elementary body of Cowdria. Two hundred and sixteen clones were obtained by limiting dilution of infected cells. The method was experimentally validated by comparing randomly amplified polymorphic DNA fingerprints from individual clones obtained from endothelial cell cultures coinfected with two different stocks of C. ruminantium.




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Antimicrob. Agents Chemother. Clin. Microbiol. Rev. Infect. Immun.
J. Clin. Microbiol. J. Virol. ALL ASM JOURNALS

Copyright © 1997 by the American Society for Microbiology. All rights reserved.