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 Citation (Scopus)

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.

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

Fingerprint

Clostridium chauvoei
Clostridium septicum
Multiplex Polymerase Chain Reaction
Ruminants
microorganisms
ruminants
Vaccines
vaccines
Enterobacter aerogenes
Flagellin
flagellin
Clostridium
DNA
Cross Reactions
Salmonella typhimurium
cross reaction
Pseudomonas aeruginosa
Salmonella Typhimurium
Genes
Staphylococcus aureus

Keywords

  • Clostridium chauvoei
  • Clostridium septicum
  • Identification
  • Multiplex PCR

ASJC Scopus subject areas

  • veterinary(all)

Cite this

PCR multiplex para identificação de isolados de Clostridium chauvoei e Clostridium septicum. / 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, Francisco A.

In: Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, Vol. 60, No. 2, 04.2008, p. 294-298.

Research output: Contribution to journalArticle

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, Francisco A. / PCR multiplex para identificação de isolados de Clostridium chauvoei e Clostridium septicum. In: Arquivo Brasileiro de Medicina Veterinaria e Zootecnia. 2008 ; Vol. 60, No. 2. pp. 294-298.
@article{90399c4715de4e6fa407186da7e20c8d,
title = "PCR multiplex para identifica{\cc}{\~a}o de isolados de Clostridium chauvoei e Clostridium septicum",
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.",
keywords = "Clostridium chauvoei, Clostridium septicum, Identification, Multiplex PCR",
author = "Assis, {R. A.} and Lobato, {F. C F} and Lobato, {Z. I P} and Camargos, {M. F.} and Nascimento, {R. A P} and Vargas, {A. P C} and Salvarani, {F. M.} and Uzal, {Francisco A}",
year = "2008",
month = "4",
doi = "10.1590/S0102-09352008000200003",
language = "Undefined/Unknown",
volume = "60",
pages = "294--298",
journal = "Arquivo Brasileiro de Medicina Veterinaria e Zootecnia",
issn = "0102-0935",
publisher = "Universidade de Minas Gerais",
number = "2",

}

TY - JOUR

T1 - PCR multiplex para identificação de isolados de Clostridium chauvoei e Clostridium septicum

AU - Assis, R. A.

AU - Lobato, F. C F

AU - Lobato, Z. I P

AU - Camargos, M. F.

AU - Nascimento, R. A P

AU - Vargas, A. P C

AU - Salvarani, F. M.

AU - Uzal, Francisco A

PY - 2008/4

Y1 - 2008/4

N2 - 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.

AB - 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.

KW - Clostridium chauvoei

KW - Clostridium septicum

KW - Identification

KW - Multiplex PCR

UR - http://www.scopus.com/inward/record.url?scp=45749129467&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=45749129467&partnerID=8YFLogxK

U2 - 10.1590/S0102-09352008000200003

DO - 10.1590/S0102-09352008000200003

M3 - Article

AN - SCOPUS:45749129467

VL - 60

SP - 294

EP - 298

JO - Arquivo Brasileiro de Medicina Veterinaria e Zootecnia

JF - Arquivo Brasileiro de Medicina Veterinaria e Zootecnia

SN - 0102-0935

IS - 2

ER -