Multiplex PCR assay for identification of Corynebacterium pseudotuberculosis from pure cultures and for rapid detection of this pathogen in clinical samples

• DOI: 10.1099/jmm.0.46997-0
• Publication Date: 2007-03-21T00:27:12Z
• Authors: Luis G. C. Pacheco; Roberta R. Pena; Thiago L. P. Castro; Fernanda A. Dorella; Robson C. Bahia; Renato Carminati; Marcílio N. L. Frota; Sérgio C. Oliveira; Roberto Meyer; Francisco S. F. A...; Anderson Miyoshi; Vasco Azevedo
• Keywords: Corynebacterium pseudotuberculosis; rpoB; Caseous lymphadenitis; Multiplex

Corynebacterium pseudotuberculosis is the aetiological agent of caseous lymphadenitis (CLA), a debilitating disease sheep and goats. Accurate diagnosis CLA primarily relies on microbiological examination, followed by biochemical identification isolates. In an effort to facilitate C. detection, multiplex PCR (mPCR) assay was developed targeting three genes this bacterium: 16S rRNA gene, rpoB pld. This method allowed efficient 40 isolates bacterium that had been identified previously testing. Analysis taxonomically related species did not generate mPCR amplification profile, thereby demonstrating assay's specificity. As little as 1 pg genomic DNA detected assay, sensitivity method. The detection limit in clinical samples estimated be 10(3) c.f.u. could directly pus from infected goats (n=56) with high diagnostic (94.6 %). significantly improves rapid supersede bacteriological culture for epidemiological CLA.