| Multiplex PCR for detection of botulinum neurotoxin-producing clostridia in clinical, food, and environmental samples. | |
| | |
MedLine Citation:
|
PMID: 19684163 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Botulinum neurotoxin (BoNT), the most toxic substance known, is produced by the spore-forming bacterium Clostridium botulinum and, in rare cases, also by some strains of Clostridium butyricum and Clostridium baratii. The standard procedure for definitive detection of BoNT-producing clostridia is a culture method combined with neurotoxin detection using a standard mouse bioassay (SMB). The SMB is highly sensitive and specific, but it is expensive and time-consuming and there are ethical concerns due to use of laboratory animals. PCR provides a rapid alternative for initial screening for BoNT-producing clostridia. In this study, a previously described multiplex PCR assay was modified to detect all type A, B, E, and F neurotoxin genes in isolated strains and in clinical, food, environmental samples. This assay includes an internal amplification control. The effectiveness of the multiplex PCR method for detecting clostridia possessing type A, B, E, and F neurotoxin genes was evaluated by direct comparison with the SMB. This method showed 100% inclusivity and 100% exclusivity when 182 BoNT-producing clostridia and 21 other bacterial strains were used. The relative accuracy of the multiplex PCR and SMB was evaluated using 532 clinical, food, and environmental samples and was estimated to be 99.2%. The multiplex PCR was also used to investigate 110 freshly collected food and environmental samples, and 4 of the 110 samples (3.6%) were positive for BoNT-encoding genes. |
| | |
Authors:
|
Dario De Medici; Fabrizio Anniballi; Gary M Wyatt; Miia Lindström; Ute Messelhäusser; Clare F Aldus; Elisabetta Delibato; Hannu Korkeala; Michael W Peck; Lucia Fenicia |
Related Documents
:
|
16619853 - Accumulation of paralytic shellfish poison (psp) and biotransformation of its component... 22765373 - Examining the occurrence of residues of flunixin meglumine in cull dairy cows by use of... 9532663 - Factors in the emergence of food borne diseases. |
Publication Detail:
|
Type: Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't Date: 2009-08-14 |
Journal Detail:
|
Title: Applied and environmental microbiology Volume: 75 ISSN: 1098-5336 ISO Abbreviation: Appl. Environ. Microbiol. Publication Date: 2009 Oct |
Date Detail:
|
Created Date: 2009-10-08 Completed Date: 2009-12-01 Revised Date: 2010-09-24 |
Medline Journal Info:
|
Nlm Unique ID: 7605801 Medline TA: Appl Environ Microbiol Country: United States |
Other Details:
|
Languages: eng Pagination: 6457-61 Citation Subset: IM |
Affiliation:
|
Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. dario.demedici@iss.it |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Base Sequence Biological Assay / statistics & numerical data Botulinum Toxins / biosynthesis*, genetics* Clostridium / genetics*, isolation & purification*, metabolism Clostridium botulinum / genetics, isolation & purification, metabolism DNA Primers / genetics DNA, Bacterial / genetics Environmental Microbiology* Food Microbiology* Genes, Bacterial Humans Mice Neurotoxins / biosynthesis*, genetics* Polymerase Chain Reaction / methods*, statistics & numerical data Sensitivity and Specificity |
| Grant Support | |
ID/Acronym/Agency:
|
//Biotechnology and Biological Sciences Research Council |
| Chemical | |
Reg. No./Substance:
|
0/Botulinum Toxins; 0/DNA Primers; 0/DNA, Bacterial; 0/Neurotoxins |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Aeromonas hydrophila AH-3 type III secretion system expression and regulatory network.
Next Document: Longitudinal study of Escherichia coli O157:H7 in a beef cattle feedlot and role of high-level shedd...