Comparison of molecular detection methods for Vibrio parahaemolyticus and Vibrio vulnificus
Pathogenic vibrios are a global concern for seafood safety and many molecular methods have been developed for their detection. This study compares several molecular methods for detection of total and pathogenic Vibrio parahaemolyticus and Vibrio vulnificus, in MPN enrichments from oysters and fish i...
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| Veröffentlicht in: | Food microbiology 2012-05, Vol.30 (1), p.105-111 |
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| creator | Jones, Jessica L. Hara-Kudo, Yukiko Krantz, Jeffrey A. Benner, Ronald A. Smith, Amy B. Dambaugh, Timothy R. Bowers, John C. DePaola, Angelo |
| description | Pathogenic vibrios are a global concern for seafood safety and many molecular methods have been developed for their detection. This study compares several molecular methods for detection of total and pathogenic Vibrio parahaemolyticus and Vibrio vulnificus, in MPN enrichments from oysters and fish intestine samples. This study employed the DuPont Qualicon BAX® System Real-Time PCR assay for detection of V. parahaemolyticus and V. vulnificus. Multiplex real-time PCR detection of total (tlh+), tdh+, and trh+V. parahaemolyticus was conducted on the Cepheid SmartCycler II. Total (rpoD) and tdh+V. parahaemolyticus were also detected using LAMP. V. vulnificus detection was performed using real-time PCR methods developed for the SmartCycler and the AB 7500 Fast. Recommended template preparations were compared to BAX® lysis samples for suitability. There was no significant difference in detection of V. parahaemolyticus and V. vulnificus using the BAX® or SmartCycler assays. The AB assay showed no difference from other methods in detection of V. vulnificus unless boiled templates were utilized. There was a significant difference in detection of tdh+V. parahaemolyticus between SmartCycler and LAMP assays unless the total (tlh+) V. parahaemolyticus gene target was omitted from the SmartCycler assay; a similar trend was observed for trh+V. parahaemolyticus.
► Many available molecular methods are suitable for detection of Vibrio vulnificus and Vibrio parahaemolyticus in foods. ► Template preparation has a significant effect on detection by molecular methods. ► Competition of within a multiplex PCR can interfere with target detection. |
| doi_str_mv | 10.1016/j.fm.2011.12.011 |
| format | Article |
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► Many available molecular methods are suitable for detection of Vibrio vulnificus and Vibrio parahaemolyticus in foods. ► Template preparation has a significant effect on detection by molecular methods. ► Competition of within a multiplex PCR can interfere with target detection.</description><subject>Colony Count, Microbial</subject><subject>Consumer Product Safety</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - isolation & purification</subject><subject>Food Contamination - analysis</subject><subject>Food Microbiology - methods</subject><subject>Loop-mediated isothermal amplification (LAMP)</subject><subject>Real-time PCR</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>Seafood - microbiology</subject><subject>Sensitivity and Specificity</subject><subject>Vibrio parahaemolyticus</subject><subject>Vibrio parahaemolyticus - genetics</subject><subject>Vibrio parahaemolyticus - isolation & purification</subject><subject>Vibrio vulnificus</subject><subject>Vibrio vulnificus - genetics</subject><subject>Vibrio vulnificus - isolation & purification</subject><issn>0740-0020</issn><issn>1095-9998</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kL1PwzAUxC0EoqWwM6FsTAm2EycxG6r4kiqxAAuD5djPqqskLnZSqf89jkrZmE6697uT3iF0TXBGMCnvNpnpMooJyQjNopygOcGcpZzz-hTNcVXgFGOKZ-gihA2OBMv5OZpRSktGOZ6jr6XrttLb4PrEmaRzLaixlT7RMIAabLQ7GNZOh8Q4n3zaxluXxIRcS4j0frBqDIns9fG2G9vemsm9RGdGtgGufnWBPp4e35cv6ert-XX5sEpVQdiQlgwUQGMKWZcNY0YXplIYGslJ2RQNAKg6r3kBGOel1lQzKkm8GWNKVjYkX6DbQ-_Wu-8RwiA6GxS0rezBjUFwUrGKFGwi8YFU3oXgwYitt530e0GwmBYVG2E6MS0qCBVRYuTmt3xsOtB_geOEEbg_ABBf3FnwIigLvQJtfVxQaGf_b_8BO9eH9w</recordid><startdate>201205</startdate><enddate>201205</enddate><creator>Jones, Jessica L.</creator><creator>Hara-Kudo, Yukiko</creator><creator>Krantz, Jeffrey A.</creator><creator>Benner, Ronald A.</creator><creator>Smith, Amy B.</creator><creator>Dambaugh, Timothy R.</creator><creator>Bowers, John C.</creator><creator>DePaola, Angelo</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201205</creationdate><title>Comparison of molecular detection methods for Vibrio parahaemolyticus and Vibrio vulnificus</title><author>Jones, Jessica L. ; Hara-Kudo, Yukiko ; Krantz, Jeffrey A. ; Benner, Ronald A. ; Smith, Amy B. ; Dambaugh, Timothy R. ; Bowers, John C. ; DePaola, Angelo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c415t-65eceebf4a86b55fd4f7c0eba916b4beeec83894e0036dd2d52a1916fff656b13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Colony Count, Microbial</topic><topic>Consumer Product Safety</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Bacterial - isolation & purification</topic><topic>Food Contamination - analysis</topic><topic>Food Microbiology - methods</topic><topic>Loop-mediated isothermal amplification (LAMP)</topic><topic>Real-time PCR</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>Seafood - microbiology</topic><topic>Sensitivity and Specificity</topic><topic>Vibrio parahaemolyticus</topic><topic>Vibrio parahaemolyticus - genetics</topic><topic>Vibrio parahaemolyticus - isolation & purification</topic><topic>Vibrio vulnificus</topic><topic>Vibrio vulnificus - genetics</topic><topic>Vibrio vulnificus - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jones, Jessica L.</creatorcontrib><creatorcontrib>Hara-Kudo, Yukiko</creatorcontrib><creatorcontrib>Krantz, Jeffrey A.</creatorcontrib><creatorcontrib>Benner, Ronald A.</creatorcontrib><creatorcontrib>Smith, Amy B.</creatorcontrib><creatorcontrib>Dambaugh, Timothy R.</creatorcontrib><creatorcontrib>Bowers, John C.</creatorcontrib><creatorcontrib>DePaola, Angelo</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Food microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jones, Jessica L.</au><au>Hara-Kudo, Yukiko</au><au>Krantz, Jeffrey A.</au><au>Benner, Ronald A.</au><au>Smith, Amy B.</au><au>Dambaugh, Timothy R.</au><au>Bowers, John C.</au><au>DePaola, Angelo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparison of molecular detection methods for Vibrio parahaemolyticus and Vibrio vulnificus</atitle><jtitle>Food microbiology</jtitle><addtitle>Food Microbiol</addtitle><date>2012-05</date><risdate>2012</risdate><volume>30</volume><issue>1</issue><spage>105</spage><epage>111</epage><pages>105-111</pages><issn>0740-0020</issn><eissn>1095-9998</eissn><abstract>Pathogenic vibrios are a global concern for seafood safety and many molecular methods have been developed for their detection. This study compares several molecular methods for detection of total and pathogenic Vibrio parahaemolyticus and Vibrio vulnificus, in MPN enrichments from oysters and fish intestine samples. This study employed the DuPont Qualicon BAX® System Real-Time PCR assay for detection of V. parahaemolyticus and V. vulnificus. Multiplex real-time PCR detection of total (tlh+), tdh+, and trh+V. parahaemolyticus was conducted on the Cepheid SmartCycler II. Total (rpoD) and tdh+V. parahaemolyticus were also detected using LAMP. V. vulnificus detection was performed using real-time PCR methods developed for the SmartCycler and the AB 7500 Fast. Recommended template preparations were compared to BAX® lysis samples for suitability. There was no significant difference in detection of V. parahaemolyticus and V. vulnificus using the BAX® or SmartCycler assays. The AB assay showed no difference from other methods in detection of V. vulnificus unless boiled templates were utilized. There was a significant difference in detection of tdh+V. parahaemolyticus between SmartCycler and LAMP assays unless the total (tlh+) V. parahaemolyticus gene target was omitted from the SmartCycler assay; a similar trend was observed for trh+V. parahaemolyticus.
► Many available molecular methods are suitable for detection of Vibrio vulnificus and Vibrio parahaemolyticus in foods. ► Template preparation has a significant effect on detection by molecular methods. ► Competition of within a multiplex PCR can interfere with target detection.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>22265290</pmid><doi>10.1016/j.fm.2011.12.011</doi><tpages>7</tpages></addata></record> |
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| subjects | Colony Count, Microbial Consumer Product Safety DNA, Bacterial - genetics DNA, Bacterial - isolation & purification Food Contamination - analysis Food Microbiology - methods Loop-mediated isothermal amplification (LAMP) Real-time PCR Real-Time Polymerase Chain Reaction Seafood - microbiology Sensitivity and Specificity Vibrio parahaemolyticus Vibrio parahaemolyticus - genetics Vibrio parahaemolyticus - isolation & purification Vibrio vulnificus Vibrio vulnificus - genetics Vibrio vulnificus - isolation & purification |
| title | Comparison of molecular detection methods for Vibrio parahaemolyticus and Vibrio vulnificus |
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