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Molecular epidemioloic analysis of Mycobacterium bovis isolates from Mexico

Feliciano Milian-SuazoCENID-Microbiología, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Km Palo Alto, D.F. CP 05110, México.

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M. D. SalmanDepartments of Environmental Health, College of Veterinary Medicine and Biological Sciences, Colorado State University, Ft Collins, CO 80523.

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William C. Black IVDepartments of Microbiology, College of Veterinary Medicine and Biological Sciences, Colorado State University, Ft Collins, CO 80523.

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Joni M. TriantisDepartments of Environmental Health, College of Veterinary Medicine and Biological Sciences, Colorado State University, Ft Collins, CO 80523.

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Carolina RamirezFCN, Universidad Autonoma de Querétaro, Querétaro, Qro, México.

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Janet B. PayeurNational Veterinary Services Laboratories, APHIS:USDA, Ames, IA 50010.

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Melaine C. TorresDepartments of Environmental Health, College of Veterinary Medicine and Biological Sciences, Colorado State University, Ft Collins, CO 80523.

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Abstract

Objective—To assess phylogenetic relationships among Mycobacterium bovis isolates by use of random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) fingerprinting and to relate genetic profiles of isolates to epidemiologic characteristics.

Animals—400 cattle with tuberculosis.

ProcedureMycobacterium bovis was isolated from various organs of cattle slaughtered in 6 geographic regions of Mexico. Most cattle were adult Holsteins from large herds that did not participate in a tuberculosis control program. Four random primers and 2 selected primers were used in RAPD-PCR fingerprinting of 88 isolates. Pairwise genetic distance between isolates was obtained and subjected to cluster analysis with bootstrapping to test for levels of support.

Results—98 different fragments were obtained; there was broad genetic diversity among isolates, and each isolate had a unique RAPD-genotype, including those originating from the same herd. Clustering by geographic location, affected organ, or severity of lesion was not detected. Linkage disequilibrium analysis suggested that M bovis was highly clonal and that mutations develop at a rapid rate among isolates.

Conclusions and Clinical Relevance—Use of RAPDPCR could not differentiate M bovis isolates by epidemiologic characteristics or identify common sources of infection. (Am J Vet Res 2000;61:90–95)

Abstract

Objective—To assess phylogenetic relationships among Mycobacterium bovis isolates by use of random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) fingerprinting and to relate genetic profiles of isolates to epidemiologic characteristics.

Animals—400 cattle with tuberculosis.

ProcedureMycobacterium bovis was isolated from various organs of cattle slaughtered in 6 geographic regions of Mexico. Most cattle were adult Holsteins from large herds that did not participate in a tuberculosis control program. Four random primers and 2 selected primers were used in RAPD-PCR fingerprinting of 88 isolates. Pairwise genetic distance between isolates was obtained and subjected to cluster analysis with bootstrapping to test for levels of support.

Results—98 different fragments were obtained; there was broad genetic diversity among isolates, and each isolate had a unique RAPD-genotype, including those originating from the same herd. Clustering by geographic location, affected organ, or severity of lesion was not detected. Linkage disequilibrium analysis suggested that M bovis was highly clonal and that mutations develop at a rapid rate among isolates.

Conclusions and Clinical Relevance—Use of RAPDPCR could not differentiate M bovis isolates by epidemiologic characteristics or identify common sources of infection. (Am J Vet Res 2000;61:90–95)