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PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species

BACKGROUND: The genus Vibrio is a diverse group of Gram-negative bacteria comprised of 74 species. Furthermore, the genus has and is expected to continue expanding with the addition of several new species annually. Consequently, it is of paramount importance to have a method which is able to reliabl...

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Autores principales: Hoffmann, Maria, Brown, Eric W, Feng, Peter CH, Keys, Christine E, Fischer, Markus, Monday, Steven R
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2856557/
https://www.ncbi.nlm.nih.gov/pubmed/20331883
http://dx.doi.org/10.1186/1471-2180-10-90
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author Hoffmann, Maria
Brown, Eric W
Feng, Peter CH
Keys, Christine E
Fischer, Markus
Monday, Steven R
author_facet Hoffmann, Maria
Brown, Eric W
Feng, Peter CH
Keys, Christine E
Fischer, Markus
Monday, Steven R
author_sort Hoffmann, Maria
collection PubMed
description BACKGROUND: The genus Vibrio is a diverse group of Gram-negative bacteria comprised of 74 species. Furthermore, the genus has and is expected to continue expanding with the addition of several new species annually. Consequently, it is of paramount importance to have a method which is able to reliably and efficiently differentiate the numerous Vibrio species. RESULTS: In this study, a novel and rapid polymerase chain reaction (PCR)-based intergenic spacer (IGS)-typing system for vibrios was developed that is based on the well-known IGS regions located between the 16S and 23S rRNA genes on the bacterial chromosome. The system was optimized to resolve heteroduplex formation as well as to take advantage of capillary gel electrophoresis technology such that reproducible analyses could be achieved in a rapid manner. System validation was achieved through testing of 69 archetypal Vibrio strains, representing 48 Vibrio species, from which an 'IGS-type' profile database was generated. These data, presented here in several cluster analyses, demonstrated successful differentiation of the 69 type strains showing that this PCR-based fingerprinting method easily discriminates bacterial strains at the species level among Vibrio. Furthermore, testing 36 strains each of V. parahaemolyticus and V. vulnificus, important food borne pathogens, isolated from a variety of geographical locations with the IGS-typing method demonstrated distinct IGS-typing patterns indicative of subspecies divergence in both populations making this technique equally useful for intraspecies differentiation, as well. CONCLUSION: This rapid, reliable and efficient IGS-typing system, especially in combination with 16S rRNA gene sequencing, has the capacity to not only discern and identify vibrios at the species level but, in some cases, at the sub-species level, as well. This procedure is particularly well-suited for preliminary species identification and, lends itself nicely to epidemiological investigations providing information more quickly than other time-honoured methods traditionally used in these types of analyses.
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spelling pubmed-28565572010-04-20 PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species Hoffmann, Maria Brown, Eric W Feng, Peter CH Keys, Christine E Fischer, Markus Monday, Steven R BMC Microbiol Methodology article BACKGROUND: The genus Vibrio is a diverse group of Gram-negative bacteria comprised of 74 species. Furthermore, the genus has and is expected to continue expanding with the addition of several new species annually. Consequently, it is of paramount importance to have a method which is able to reliably and efficiently differentiate the numerous Vibrio species. RESULTS: In this study, a novel and rapid polymerase chain reaction (PCR)-based intergenic spacer (IGS)-typing system for vibrios was developed that is based on the well-known IGS regions located between the 16S and 23S rRNA genes on the bacterial chromosome. The system was optimized to resolve heteroduplex formation as well as to take advantage of capillary gel electrophoresis technology such that reproducible analyses could be achieved in a rapid manner. System validation was achieved through testing of 69 archetypal Vibrio strains, representing 48 Vibrio species, from which an 'IGS-type' profile database was generated. These data, presented here in several cluster analyses, demonstrated successful differentiation of the 69 type strains showing that this PCR-based fingerprinting method easily discriminates bacterial strains at the species level among Vibrio. Furthermore, testing 36 strains each of V. parahaemolyticus and V. vulnificus, important food borne pathogens, isolated from a variety of geographical locations with the IGS-typing method demonstrated distinct IGS-typing patterns indicative of subspecies divergence in both populations making this technique equally useful for intraspecies differentiation, as well. CONCLUSION: This rapid, reliable and efficient IGS-typing system, especially in combination with 16S rRNA gene sequencing, has the capacity to not only discern and identify vibrios at the species level but, in some cases, at the sub-species level, as well. This procedure is particularly well-suited for preliminary species identification and, lends itself nicely to epidemiological investigations providing information more quickly than other time-honoured methods traditionally used in these types of analyses. BioMed Central 2010-03-23 /pmc/articles/PMC2856557/ /pubmed/20331883 http://dx.doi.org/10.1186/1471-2180-10-90 Text en Copyright ©2010 Hoffmann et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology article
Hoffmann, Maria
Brown, Eric W
Feng, Peter CH
Keys, Christine E
Fischer, Markus
Monday, Steven R
PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species
title PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species
title_full PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species
title_fullStr PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species
title_full_unstemmed PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species
title_short PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species
title_sort pcr-based method for targeting 16s-23s rrna intergenic spacer regions among vibrio species
topic Methodology article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2856557/
https://www.ncbi.nlm.nih.gov/pubmed/20331883
http://dx.doi.org/10.1186/1471-2180-10-90
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