Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic
ABSTRACT Pulsed-field gel electrophoresis (PFGE), randomly amplified polymorphic DNA (RAPD) analysis, and automated ribotyping were compared for epidemiological typing of Clostridium botulinum type E using clinical and food isolates associated with four botulism outbreaks occurring in the Canadian A...
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Online Access: | http://dx.doi.org/10.1128/jcm.44.5.1635-1644.2006 https://journals.asm.org/doi/pdf/10.1128/JCM.44.5.1635-1644.2006 |
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crasmicro:10.1128/jcm.44.5.1635-1644.2006 2023-11-05T03:39:18+01:00 Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic Leclair, Daniel Pagotto, Franco Farber, Jeffrey M. Cadieux, Brigitte Austin, John W. 2006 http://dx.doi.org/10.1128/jcm.44.5.1635-1644.2006 https://journals.asm.org/doi/pdf/10.1128/JCM.44.5.1635-1644.2006 en eng American Society for Microbiology https://journals.asm.org/non-commercial-tdm-license Journal of Clinical Microbiology volume 44, issue 5, page 1635-1644 ISSN 0095-1137 1098-660X Microbiology (medical) journal-article 2006 crasmicro https://doi.org/10.1128/jcm.44.5.1635-1644.2006 2023-10-09T16:01:42Z ABSTRACT Pulsed-field gel electrophoresis (PFGE), randomly amplified polymorphic DNA (RAPD) analysis, and automated ribotyping were compared for epidemiological typing of Clostridium botulinum type E using clinical and food isolates associated with four botulism outbreaks occurring in the Canadian Arctic. All type E strains previously untypeable by PFGE, even with the use of a formaldehyde fixation step, could be typed by the addition of 50 μM thiourea to the electrophoresis running buffer. Digestion with SmaI or XhoI followed by PFGE was used to link food and clinical isolates from four different type E botulism outbreaks and differentiate them from among 39 group II strains. Strain differentiation was unsuccessful with the automated ribotyping system, producing a single characteristic EcoRI fingerprint common to all group II strains. RAPD analysis of C. botulinum group II strains was not consistently reproducible with primer OPJ-6 or OPJ-13, apparently discriminating between epidemiologically related strains. A modified PFGE protocol was judged to be the most useful method for typing epidemiologically related C. botulinum type E strains, based on its ability to type all strains reproducibly and with an adequate level of discrimination. Article in Journal/Newspaper Arctic ASM Journals (American Society for Microbiology - via Crossref) Journal of Clinical Microbiology 44 5 1635 1644 |
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ASM Journals (American Society for Microbiology - via Crossref) |
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English |
topic |
Microbiology (medical) |
spellingShingle |
Microbiology (medical) Leclair, Daniel Pagotto, Franco Farber, Jeffrey M. Cadieux, Brigitte Austin, John W. Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic |
topic_facet |
Microbiology (medical) |
description |
ABSTRACT Pulsed-field gel electrophoresis (PFGE), randomly amplified polymorphic DNA (RAPD) analysis, and automated ribotyping were compared for epidemiological typing of Clostridium botulinum type E using clinical and food isolates associated with four botulism outbreaks occurring in the Canadian Arctic. All type E strains previously untypeable by PFGE, even with the use of a formaldehyde fixation step, could be typed by the addition of 50 μM thiourea to the electrophoresis running buffer. Digestion with SmaI or XhoI followed by PFGE was used to link food and clinical isolates from four different type E botulism outbreaks and differentiate them from among 39 group II strains. Strain differentiation was unsuccessful with the automated ribotyping system, producing a single characteristic EcoRI fingerprint common to all group II strains. RAPD analysis of C. botulinum group II strains was not consistently reproducible with primer OPJ-6 or OPJ-13, apparently discriminating between epidemiologically related strains. A modified PFGE protocol was judged to be the most useful method for typing epidemiologically related C. botulinum type E strains, based on its ability to type all strains reproducibly and with an adequate level of discrimination. |
format |
Article in Journal/Newspaper |
author |
Leclair, Daniel Pagotto, Franco Farber, Jeffrey M. Cadieux, Brigitte Austin, John W. |
author_facet |
Leclair, Daniel Pagotto, Franco Farber, Jeffrey M. Cadieux, Brigitte Austin, John W. |
author_sort |
Leclair, Daniel |
title |
Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic |
title_short |
Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic |
title_full |
Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic |
title_fullStr |
Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic |
title_full_unstemmed |
Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic |
title_sort |
comparison of dna fingerprinting methods for use in investigation of type e botulism outbreaks in the canadian arctic |
publisher |
American Society for Microbiology |
publishDate |
2006 |
url |
http://dx.doi.org/10.1128/jcm.44.5.1635-1644.2006 https://journals.asm.org/doi/pdf/10.1128/JCM.44.5.1635-1644.2006 |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Clinical Microbiology volume 44, issue 5, page 1635-1644 ISSN 0095-1137 1098-660X |
op_rights |
https://journals.asm.org/non-commercial-tdm-license |
op_doi |
https://doi.org/10.1128/jcm.44.5.1635-1644.2006 |
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Journal of Clinical Microbiology |
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44 |
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5 |
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1635 |
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1644 |
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1781695112630239232 |