PCR multiplex para identificação de isolados de Clostridium chauvoei e Clostridium septicum

Translated title of the contribution: Multiplex PCR for identification of Clostridium chauvoei and Clostridium septicum

R. A. Assis, F. C F Lobato, Z. I P Lobato, M. F. Camargos, R. A P Nascimento, A. P C Vargas, F. M. Salvarani, Francisco A Uzal

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Multiplex PCR was optimized to detect Clostridium chauvoei and Clostridium septicum in pure cultures. In each reaction, a pair of primers for a specific segment of the flagellin gene of C. chauvoei and a pair of primers for a specific segment of alpha toxin gene of C. septicum were employed. Reference strains of both microorganisms were used as control. The multiplex PCR was evaluated by testing 16 clinical isolates of C. chauvoei from ruminants, 15 clinical isolates of C. septicum from ruminants and, four vaccine strains of each one of these agents. Reference strains of both microorganisms were used as control. To evaluate the specificity, genomic DNA of the following microorganisms was used: C. sordellii, C. novyi type A, C. novyi type B, C. perfringens type A, C. haemolyticum, C. botulinum type D, Pseudomonas aeruginosa, Staphylococcus aureus, Enterobacter aerogenes, Escherichia coli, and Salmonella typhimurium. All the isolates and vaccine strains of C. chauvoei and C. septicum were positive by PCR assay and cross reactions were not observed with the other species of clostridia, the other bacterial species or amongst both investigated agents. The smallest concentrations of DNA detected from C. chauvoei and C. septicum were 45pg/μl and 30pg/μl, respectively. The multiplex PCR was useful for the specific identification of C. chauvoei and C. septicum in pure cultures.

Translated title of the contributionMultiplex PCR for identification of Clostridium chauvoei and Clostridium septicum
Original languageUndefined/Unknown
Pages (from-to)294-298
Number of pages5
JournalArquivo Brasileiro de Medicina Veterinaria e Zootecnia
Volume60
Issue number2
DOIs
StatePublished - Apr 2008

Keywords

  • Clostridium chauvoei
  • Clostridium septicum
  • Identification
  • Multiplex PCR

ASJC Scopus subject areas

  • veterinary(all)

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    Assis, R. A., Lobato, F. C. F., Lobato, Z. I. P., Camargos, M. F., Nascimento, R. A. P., Vargas, A. P. C., Salvarani, F. M., & Uzal, F. A. (2008). PCR multiplex para identificação de isolados de Clostridium chauvoei e Clostridium septicum. Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, 60(2), 294-298. https://doi.org/10.1590/S0102-09352008000200003