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Título: | Multiplex PCR assay for identification of Corynebacterium pseudotuberculosis from pure cultures and for rapid detection of this pathogen in clinical samples. |
Autor: | PACHECO, L. G. C.![]() ![]() PENA, R. R. ![]() ![]() CASTRO, T. L. P. ![]() ![]() DORELLA, F. A. ![]() ![]() BAHIA, R. C. ![]() ![]() CARMINATI, R. ![]() ![]() FROTA, M. N. L. ![]() ![]() OLIVEIRA, S. C. ![]() ![]() MEYER, R. ![]() ![]() ALVES, F. S. F. ![]() ![]() MIYOSHI, A. ![]() ![]() AZEVEDO, V. ![]() ![]() |
Afiliación: | LUIS G. C. PACHECO, UNIVERSIDADE FEDERAL DE MINAS GERAIS ROBERTA R. PENA, UNIVERSIDADE FEDERAL DE MINAS GERAIS UNIVERSIDADE FEDERAL DE MINAS GERAIS UNIVERSIDADE FEDERAL DE MINAS GERAIS ROBSON C. BAHIA, UNIVERSIDADE FEDERAL DA BAHIA RENATO CARMINATI, UNIVERSIDADE FEDERAL DA BAHIA MAURÍCIO N. L. FROTA, UNIVERSIDADE ESTADUAL VALE DO ACARAÚ SÉRGIO C. OLIVEIRA, UNIVERSIDADE FEDERAL DE MINAS GERAIS ROBERTO MEYER, UNIVERSIDADE FEDERAL DA BAHIA FRANCISCO SELMO FERNANDES ALVES, CNPC ANDERSON MIYOSHI, UNIVERSIDADE FEDERAL DE MINAS GERAIS VASCO AZEVEDO, UNIVERSIDADE FEDERAL DE MINAS GERAIS. |
Año: | 2007 |
Referencia: | Journal of Medical Microbiology, v. 56, n. 4, p. 480-486, Apr. 2007. |
Descripción: | Abstract: Corynebacterium pseudotuberculosis is the aetiological agent of caseous lymphadenitis (CLA), a debilitating disease of sheep and goats. Accurate diagnosis of CLA primarily relies on microbiological examination, followed by biochemical identification of isolates. In an effort to facilitate C. pseudotuberculosis detection, a multiplex PCR (mPCR) assay was developed targeting three genes of this bacterium: the 16S rRNA gene, rpoB and pld. This method allowed efficient identification of 40 isolates of this bacterium that had been identified previously by biochemical testing. Analysis of taxonomically related species did not generate the C. pseudotuberculosis mPCR amplification profile, thereby demonstrating the assay's specificity. As little as 1 pg of C. pseudotuberculosis genomic DNA was detected by this mPCR assay, demonstrating the sensitivity of the method. The detection limit in clinical samples was estimated to be 103 c.f.u. C. pseudotuberculosis could be detected directly in pus samples from infected sheep and goats (n=56) with a high diagnostic sensitivity (94.6 %). The developed assay significantly improves rapid C. pseudotuberculosis detection and could supersede bacteriological culture for microbiological and epidemiological diagnosis of CLA. |
Thesagro: | Caprino Ovino Corynebacterium Pseudotuberculosis Linfadenite Caseosa |
NAL Thesaurus: | Caseous lymphadenitis Sheep diseases Polymerase chain reaction Lymph nodes |
Palabras clave: | PCR CLA Base eequence Molecular sequence data Corynebacterium Infections RNA bacterial Bacterial diseases |
DOI: | 10.1099/jmm.0.46997-0 |
Tipo de Material: | Artigo de periódico |
Acceso: | openAccess |
Aparece en las colecciones: | Artigo em periódico indexado (CNPC)![]() ![]() |
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APIMultiplexPCRassay.pdf | 188.03 kB | Adobe PDF | ![]() Visualizar/Abrir |