Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing
ABSTRACT A total of 41 Clostridium botulinum serotype E strains from different geographic regions, including Canada, Denmark, Finland, France, Greenland, Japan, and the United States, were compared by multilocus sequence typing (MLST), amplified fragment length polymorphism (AFLP) analysis, variable...
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Online Access: | http://dx.doi.org/10.1128/aem.05155-11 https://journals.asm.org/doi/pdf/10.1128/AEM.05155-11 |
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crasmicro:10.1128/aem.05155-11 2024-09-15T18:10:00+00:00 Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing Macdonald, Thomas E. Helma, Charles H. Shou, Yulin Valdez, Yolanda E. Ticknor, Lawrence O. Foley, Brian T. Davis, Stephen W. Hannett, George E. Kelly-Cirino, Cassandra D. Barash, Jason R. Arnon, Stephen S. Lindström, Miia Korkeala, Hannu Smith, Leonard A. Smith, Theresa J. Hill, Karen K. 2011 http://dx.doi.org/10.1128/aem.05155-11 https://journals.asm.org/doi/pdf/10.1128/AEM.05155-11 en eng American Society for Microbiology https://journals.asm.org/non-commercial-tdm-license Applied and Environmental Microbiology volume 77, issue 24, page 8625-8634 ISSN 0099-2240 1098-5336 journal-article 2011 crasmicro https://doi.org/10.1128/aem.05155-11 2024-07-29T04:08:44Z ABSTRACT A total of 41 Clostridium botulinum serotype E strains from different geographic regions, including Canada, Denmark, Finland, France, Greenland, Japan, and the United States, were compared by multilocus sequence typing (MLST), amplified fragment length polymorphism (AFLP) analysis, variable-number tandem-repeat (VNTR) analysis, and botulinum neurotoxin ( bont ) E gene sequencing. The strains, representing environmental, food-borne, and infant botulism samples collected from 1932 to 2007, were analyzed to compare serotype E strains from different geographic regions and types of botulism and to determine whether each of the strains contained the transposon-associated recombinase rarA , involved with bont / E insertion. MLST examination using 15 genes clustered the strains into several clades, with most members within a cluster sharing the same BoNT/E subtype (BoNT/E1, E2, E3, or E6). Sequencing of the bont / E gene identified two new variants (E7, E8) that showed regions of recombination with other E subtypes. The AFLP dendrogram clustered the 41 strains similarly to the MLST dendrogram. Strains that could not be differentiated by AFLP, MLST, or bont gene sequencing were further examined using three VNTR regions. Both intact and split rarA genes were amplified by PCR in each of the strains, and their identities were confirmed in 11 strains by amplicon sequencing. The findings suggest that (i) the C. botulinum serotype E strains result from the targeted insertion of the bont / E gene into genetically conserved bacteria and (ii) recombination events (not random mutations) within bont / E result in toxin variants or subtypes within strains. Article in Journal/Newspaper Greenland ASM Journals (American Society for Microbiology) Applied and Environmental Microbiology 77 24 8625 8634 |
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ASM Journals (American Society for Microbiology) |
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crasmicro |
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English |
description |
ABSTRACT A total of 41 Clostridium botulinum serotype E strains from different geographic regions, including Canada, Denmark, Finland, France, Greenland, Japan, and the United States, were compared by multilocus sequence typing (MLST), amplified fragment length polymorphism (AFLP) analysis, variable-number tandem-repeat (VNTR) analysis, and botulinum neurotoxin ( bont ) E gene sequencing. The strains, representing environmental, food-borne, and infant botulism samples collected from 1932 to 2007, were analyzed to compare serotype E strains from different geographic regions and types of botulism and to determine whether each of the strains contained the transposon-associated recombinase rarA , involved with bont / E insertion. MLST examination using 15 genes clustered the strains into several clades, with most members within a cluster sharing the same BoNT/E subtype (BoNT/E1, E2, E3, or E6). Sequencing of the bont / E gene identified two new variants (E7, E8) that showed regions of recombination with other E subtypes. The AFLP dendrogram clustered the 41 strains similarly to the MLST dendrogram. Strains that could not be differentiated by AFLP, MLST, or bont gene sequencing were further examined using three VNTR regions. Both intact and split rarA genes were amplified by PCR in each of the strains, and their identities were confirmed in 11 strains by amplicon sequencing. The findings suggest that (i) the C. botulinum serotype E strains result from the targeted insertion of the bont / E gene into genetically conserved bacteria and (ii) recombination events (not random mutations) within bont / E result in toxin variants or subtypes within strains. |
format |
Article in Journal/Newspaper |
author |
Macdonald, Thomas E. Helma, Charles H. Shou, Yulin Valdez, Yolanda E. Ticknor, Lawrence O. Foley, Brian T. Davis, Stephen W. Hannett, George E. Kelly-Cirino, Cassandra D. Barash, Jason R. Arnon, Stephen S. Lindström, Miia Korkeala, Hannu Smith, Leonard A. Smith, Theresa J. Hill, Karen K. |
spellingShingle |
Macdonald, Thomas E. Helma, Charles H. Shou, Yulin Valdez, Yolanda E. Ticknor, Lawrence O. Foley, Brian T. Davis, Stephen W. Hannett, George E. Kelly-Cirino, Cassandra D. Barash, Jason R. Arnon, Stephen S. Lindström, Miia Korkeala, Hannu Smith, Leonard A. Smith, Theresa J. Hill, Karen K. Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing |
author_facet |
Macdonald, Thomas E. Helma, Charles H. Shou, Yulin Valdez, Yolanda E. Ticknor, Lawrence O. Foley, Brian T. Davis, Stephen W. Hannett, George E. Kelly-Cirino, Cassandra D. Barash, Jason R. Arnon, Stephen S. Lindström, Miia Korkeala, Hannu Smith, Leonard A. Smith, Theresa J. Hill, Karen K. |
author_sort |
Macdonald, Thomas E. |
title |
Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing |
title_short |
Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing |
title_full |
Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing |
title_fullStr |
Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing |
title_full_unstemmed |
Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing |
title_sort |
analysis of clostridium botulinum serotype e strains by using multilocus sequence typing, amplified fragment length polymorphism, variable-number tandem-repeat analysis, and botulinum neurotoxin gene sequencing |
publisher |
American Society for Microbiology |
publishDate |
2011 |
url |
http://dx.doi.org/10.1128/aem.05155-11 https://journals.asm.org/doi/pdf/10.1128/AEM.05155-11 |
genre |
Greenland |
genre_facet |
Greenland |
op_source |
Applied and Environmental Microbiology volume 77, issue 24, page 8625-8634 ISSN 0099-2240 1098-5336 |
op_rights |
https://journals.asm.org/non-commercial-tdm-license |
op_doi |
https://doi.org/10.1128/aem.05155-11 |
container_title |
Applied and Environmental Microbiology |
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77 |
container_issue |
24 |
container_start_page |
8625 |
op_container_end_page |
8634 |
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1810447603308953600 |