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Evaluation of IS200-PCR and comparison with other molecular markers To trace Salmonella enterica subsp. enterica serotype typhimurium bovine isolates from farm to meat - PubMed

Comparative Study

Evaluation of IS200-PCR and comparison with other molecular markers To trace Salmonella enterica subsp. enterica serotype typhimurium bovine isolates from farm to meat

Y Millemann et al. J Clin Microbiol. 2000 Jun.

Abstract

A procedure that uses an original molecular marker (IS200-PCR) and that is based on the amplification of DNA with outward-facing primers complementary to each end of IS200 has been evaluated with a collection of 85 Salmonella enterica subsp. enterica serotype Typhimurium isolates. These strains were isolated from a group of 10 cows at different stages: during transportation between the farm and the slaughterhouse, on the slaughter line, from the environment, and from the final product (ground beef). The 85 isolates were characterized by their antibiotic resistance patterns and were compared by IS200-PCR and by use of four other genotypic markers. Those markers included restriction profiles for 16S and 23S rRNA (ribotypes) and amplification profiles obtained by different approaches: random amplified polymorphic DNA analysis, enterobacterial repetitive intergenic consensus PCR, and PCR ribotyping. The results of the IS200-PCR were in accordance with those of other molecular typing methods for this collection of isolates. Five different genotypes were found, which made it possible to refine the hypotheses on transmission obtained from phenotypic results. The genotyping results indicated the massive contamination of the whole group of animals and of the environment by one clonal strain originally recovered from one cow that excreted the strain. On the other hand, a few animals and their environment appeared to be simultaneously contaminated with genetically different strains.

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Figures

FIG. 1
FIG. 1

IS200-PCR amplification profiles of S. enterica subsp. enterica serotype Typhimurium isolates. Lanes 1 to 8, reference strains F98, BN8301, BN8501, BN91A1, BN91C1, IP5210, IP5858, and IP6062, respectively; lane 9, molecular size Marker VIII (Boehringer Mannheim); lanes 10 to 17, isolates LV3, LV26, LV33, LV55, LV30, LV40, LV50, and LV60, respectively.

FIG. 2
FIG. 2

PCR-ribotypes of the Salmonella strains studied. Lanes 1 to 8, reference strains F98, BN8301, BN8501, BN91A1, BN91C1, IP5210, IP5858, and IP6062, respectively; lane M, molecular weight Marker VIII (Boehringer Mannheim); lanes 10 to 14, isolates LV3, LV6, LV26, LV33, and LV55, respectively.

FIG. 3
FIG. 3

Dendrogram showing the results of the ascending hierarchical classification based on PCR types of the 85 bovine S. enterica subsp. enterica serotype Typhimurium isolates presented in Table 1.

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