PCR detection of Clostridium chauvoei in pure cultures and in formalin-fixed, paraffin-embedded tissues

Uzal, FA, Hugenholtz, P, Blackall, LL, Petray, S, Moss, S, Assis, RA, Miyakawa, MF and Carloni, G (2003) PCR detection of Clostridium chauvoei in pure cultures and in formalin-fixed, paraffin-embedded tissues. Veterinary Microbiology, 91 2-3: 239-248. doi:10.1016/S0378-1135(02)00291-2


Author Uzal, FA
Hugenholtz, P
Blackall, LL
Petray, S
Moss, S
Assis, RA
Miyakawa, MF
Carloni, G
Title PCR detection of Clostridium chauvoei in pure cultures and in formalin-fixed, paraffin-embedded tissues
Journal name Veterinary Microbiology   Check publisher's open access policy
ISSN 0378-1135
Publication date 2003-02-01
Year available 2003
Sub-type Article (original research)
DOI 10.1016/S0378-1135(02)00291-2
Open Access Status DOI
Volume 91
Issue 2-3
Start page 239
End page 248
Total pages 10
Place of publication AMSTERDAM
Publisher ELSEVIER SCIENCE BV
Language eng
Subject 2404 Microbiology
3400 Veterinary
Abstract The polymerase chain reaction (PCR) was used to amplify specific segments of the 16S ribosomal RNA gene of Clostridium chauvoei, a major pathogen of ruminants. Three sets of primers were used to produce amplicons of 159, 836 and 959 base pairs (bp), respectively. The PCR was evaluated by testing clinically important strains of Clostridium, including 21 strains of C. chauvoei, five strains each of Clostridium septicum and Clostridium perfringens and two strains each of Clostridium novyi, Clostridium histolyticum and Clostridium sordellii. Both purified DNA and biomass from pure cultures of each of these microorganisms were evaluated as templates in the PCR. In addition, extracts of formalin-fixed, paraffin-embedded tissues of eight sheep experimentally inoculated with C. chauvoei or C septicum (four animals each) were also tested by the PCR using the three sets of primers. Purified DNA template of all C. chauvoei strains produced PCR amplicons of the expected size for all three primer pairs. However, when biomass from pure cultures of C. chauvoei or tissue extracts were used as templates, only the primer pair designed to produce the 159 bp amplicon gave consistently positive results. No positive results were obtained with any primer pair when purified DNA or biomass from pure cultures of non-target clostridial species were used as templates. Therefore, the PCR primer sets appear to be very specific for identifying C chauvoei in both cultures and tissues. (C) 2002 Elsevier Science B.V. All rights reserved.
Keyword black leg
Clostridium chauvoei
diagnosis
malignant oedema
Pcr
Identification
Sequence
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: ResearcherID Downloads - Archived
 
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