Cargando…

Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant

Listeria innocua is an important hygiene indicator bacterium in food industries because it behaves similar to Listeria monocytogenes, which is pathogenic to humans. PFGE is often used to characterize bacterial strains and to track contamination source. However, because PFGE is an expensive, complica...

Descripción completa

Detalles Bibliográficos
Autores principales: Takahashi, Hajime, Ohshima, Chihiro, Nakagawa, Miku, Thanatsang, Krittaporn, Phraephaisarn, Chirapiphat, Chaturongkasumrit, Yuphakhun, Keeratipibul, Suwimon, Kuda, Takashi, Kimura, Bon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4157758/
https://www.ncbi.nlm.nih.gov/pubmed/25198191
http://dx.doi.org/10.1371/journal.pone.0105803
_version_ 1782333923227860992
author Takahashi, Hajime
Ohshima, Chihiro
Nakagawa, Miku
Thanatsang, Krittaporn
Phraephaisarn, Chirapiphat
Chaturongkasumrit, Yuphakhun
Keeratipibul, Suwimon
Kuda, Takashi
Kimura, Bon
author_facet Takahashi, Hajime
Ohshima, Chihiro
Nakagawa, Miku
Thanatsang, Krittaporn
Phraephaisarn, Chirapiphat
Chaturongkasumrit, Yuphakhun
Keeratipibul, Suwimon
Kuda, Takashi
Kimura, Bon
author_sort Takahashi, Hajime
collection PubMed
description Listeria innocua is an important hygiene indicator bacterium in food industries because it behaves similar to Listeria monocytogenes, which is pathogenic to humans. PFGE is often used to characterize bacterial strains and to track contamination source. However, because PFGE is an expensive, complicated, time-consuming protocol, and poses difficulty in data sharing, development of a new typing method is necessary. MLVA is a technique that identifies bacterial strains on the basis of the number of tandem repeats present in the genome varies depending on the strains. MLVA has gained attention due to its high reproducibility and ease of data sharing. In this study, we developed a MLVA protocol to assess L. innocua and evaluated it by tracking the contamination source of L. innocua in an actual food manufacturing factory by typing the bacterial strains isolated from the factory. Three VNTR regions of the L. innocua genome were chosen for use in the MLVA. The number of repeat units in each VNTR region was calculated based on the results of PCR product analysis using capillary electrophoresis (CE). The calculated number of repetitions was compared with the results of the gene sequence analysis to demonstrate the accuracy of the CE repeat number analysis. The developed technique was evaluated using 60 L. innocua strains isolated from a food factory. These 60 strains were classified into 11 patterns using MLVA. Many of the strains were classified into ST-6, revealing that this MLVA strain type can contaminate each manufacturing process in the factory. The MLVA protocol developed in this study for L. innocua allowed rapid and easy analysis through the use of CE. This technique was found to be very useful in hygiene control in factories because it allowed us to track contamination sources and provided information regarding whether the bacteria were present in the factories.
format Online
Article
Text
id pubmed-4157758
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-41577582014-09-09 Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant Takahashi, Hajime Ohshima, Chihiro Nakagawa, Miku Thanatsang, Krittaporn Phraephaisarn, Chirapiphat Chaturongkasumrit, Yuphakhun Keeratipibul, Suwimon Kuda, Takashi Kimura, Bon PLoS One Research Article Listeria innocua is an important hygiene indicator bacterium in food industries because it behaves similar to Listeria monocytogenes, which is pathogenic to humans. PFGE is often used to characterize bacterial strains and to track contamination source. However, because PFGE is an expensive, complicated, time-consuming protocol, and poses difficulty in data sharing, development of a new typing method is necessary. MLVA is a technique that identifies bacterial strains on the basis of the number of tandem repeats present in the genome varies depending on the strains. MLVA has gained attention due to its high reproducibility and ease of data sharing. In this study, we developed a MLVA protocol to assess L. innocua and evaluated it by tracking the contamination source of L. innocua in an actual food manufacturing factory by typing the bacterial strains isolated from the factory. Three VNTR regions of the L. innocua genome were chosen for use in the MLVA. The number of repeat units in each VNTR region was calculated based on the results of PCR product analysis using capillary electrophoresis (CE). The calculated number of repetitions was compared with the results of the gene sequence analysis to demonstrate the accuracy of the CE repeat number analysis. The developed technique was evaluated using 60 L. innocua strains isolated from a food factory. These 60 strains were classified into 11 patterns using MLVA. Many of the strains were classified into ST-6, revealing that this MLVA strain type can contaminate each manufacturing process in the factory. The MLVA protocol developed in this study for L. innocua allowed rapid and easy analysis through the use of CE. This technique was found to be very useful in hygiene control in factories because it allowed us to track contamination sources and provided information regarding whether the bacteria were present in the factories. Public Library of Science 2014-09-08 /pmc/articles/PMC4157758/ /pubmed/25198191 http://dx.doi.org/10.1371/journal.pone.0105803 Text en © 2014 Takahashi et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Takahashi, Hajime
Ohshima, Chihiro
Nakagawa, Miku
Thanatsang, Krittaporn
Phraephaisarn, Chirapiphat
Chaturongkasumrit, Yuphakhun
Keeratipibul, Suwimon
Kuda, Takashi
Kimura, Bon
Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant
title Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant
title_full Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant
title_fullStr Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant
title_full_unstemmed Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant
title_short Development of New Multilocus Variable Number of Tandem Repeat Analysis (MLVA) for Listeria innocua and Its Application in a Food Processing Plant
title_sort development of new multilocus variable number of tandem repeat analysis (mlva) for listeria innocua and its application in a food processing plant
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4157758/
https://www.ncbi.nlm.nih.gov/pubmed/25198191
http://dx.doi.org/10.1371/journal.pone.0105803
work_keys_str_mv AT takahashihajime developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT ohshimachihiro developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT nakagawamiku developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT thanatsangkrittaporn developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT phraephaisarnchirapiphat developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT chaturongkasumrityuphakhun developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT keeratipibulsuwimon developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT kudatakashi developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant
AT kimurabon developmentofnewmultilocusvariablenumberoftandemrepeatanalysismlvaforlisteriainnocuaanditsapplicationinafoodprocessingplant