Identification of Corynebacterium pseudotuberculosis from sheep by PCR-restriction analysis using the RNA polymerase β-subunit gene (rpoB)

Caseous lymphadenitis, caused by Corynebacterium pseudotuberculosis, has a high prevalence in many regions of the world, including Argentina and Brazil. A polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method for the identification of this microorganism was designed ba...

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Bibliographic Details
Published in:Research in veterinary science 2012-04, Vol.92 (2), p.202-206
Main Authors: Pavan, M.E., Robles, C., Cairó, F.M., Marcellino, R., Pettinari, M.J.
Format: Article
Language:English
Subjects:
Animals
Arcanobacterium pyogenes
Caseous lymphadenitis
Corynebacterium
Corynebacterium Infections - diagnosis
Corynebacterium Infections - microbiology
Corynebacterium Infections - veterinary
Corynebacterium pseudotuberculosis
Corynebacterium pseudotuberculosis - enzymology
Corynebacterium pseudotuberculosis - genetics
DNA-Directed RNA Polymerases - genetics
Polymorphism, Restriction Fragment Length - genetics
Reverse Transcriptase Polymerase Chain Reaction - methods
Reverse Transcriptase Polymerase Chain Reaction - veterinary
rpoB PCR–RFLP
Sequence Analysis - veterinary
Sheep - microbiology
Sheep Diseases - diagnosis
Sheep Diseases - microbiology
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Summary:Caseous lymphadenitis, caused by Corynebacterium pseudotuberculosis, has a high prevalence in many regions of the world, including Argentina and Brazil. A polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method for the identification of this microorganism was designed based on the hypervariable region of the polymorphic RNA polymerase β-subunit gene (rpoB). All available CorynebacteriumrpoB sequences were analyzed by computer-assisted restriction analysis. The rpoB PCR–RFLP pattern predicted by using endonucleases MseI and StuI clearly differentiated C. pseudotuberculosis from sixty-one other Corynebacterium species. This method was successfully applied to identify twelve wild C. pseudotuberculosis ovine isolates and one caprine isolate. It was also used to differentiate C. pseudotuberculosis from Arcanobacterium pyogenes, an ovine pathogen with similar clinical characteristics. These results indicate that this new molecular method can be used for the reliable identification of the pathogen, essential for the timely detection of infected animals and for epidemiological studies.
ISSN:0034-5288
1532-2661
DOI:10.1016/j.rvsc.2011.02.007