A New Rapid Real-Time PCR Method for Detection of Listeria monocytogenes Targeting the hlyA Gene
In this study, primer sets of L. monocytogenes virulence gene hlyA were designed for the L. monocytogenes-specific PCR assay. To evaluate the performance of these primer sets, the detection sensitivity and specificity were examined and the most suitable primer set was selected. Subsequently, it was...
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| Veröffentlicht in: | FOOD SCIENCE AND TECHNOLOGY RESEARCH 2012, Vol.18(1), pp.47-57 |
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| creator | LIU, Pei MIZUE, Hiromi FUJIHARA, Kumiko KOBAYASHI, Hiroshi KAMIKADO, Hideaki TANAKA, Takashi HONJOH, Ken-ichi MIYAMOTO, Takahisa |
| description | In this study, primer sets of L. monocytogenes virulence gene hlyA were designed for the L. monocytogenes-specific PCR assay. To evaluate the performance of these primer sets, the detection sensitivity and specificity were examined and the most suitable primer set was selected. Subsequently, it was subjected to compare the performance for detection of L. monocytogenes with commercially available kits (TaKaRa detection kit and ABI detection kit) on DNA level. Results of real-time PCR showed that the efficiency of this new primer set was 101.6% while TaKaRa and ABI detection kit were 101.1% and 107.4%, respectively. The detection sensitivity of all three methods was equivalent to less than 1 copy per reaction using purified genomic DNA as template. Detection specificity were 100% when testing L. monocytogenes isolates, and for other Listeria isolates were 15.4%, 76.9% and 100% by the method using hlyA L. monocytogenes detection primer set 7, TaKaRa and ABI detection kit, respectively. In summary, this new PCR detection method is relatively sensitive and more specific to L. monocytogenes under certain conditions, could serve as a rapid detection method and has the potential for detection of L. monocytogenes from contaminated food. |
| doi_str_mv | 10.3136/fstr.18.47 |
| format | Article |
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To evaluate the performance of these primer sets, the detection sensitivity and specificity were examined and the most suitable primer set was selected. Subsequently, it was subjected to compare the performance for detection of L. monocytogenes with commercially available kits (TaKaRa detection kit and ABI detection kit) on DNA level. Results of real-time PCR showed that the efficiency of this new primer set was 101.6% while TaKaRa and ABI detection kit were 101.1% and 107.4%, respectively. The detection sensitivity of all three methods was equivalent to less than 1 copy per reaction using purified genomic DNA as template. Detection specificity were 100% when testing L. monocytogenes isolates, and for other Listeria isolates were 15.4%, 76.9% and 100% by the method using hlyA L. monocytogenes detection primer set 7, TaKaRa and ABI detection kit, respectively. In summary, this new PCR detection method is relatively sensitive and more specific to L. monocytogenes under certain conditions, could serve as a rapid detection method and has the potential for detection of L. monocytogenes from contaminated food.</description><identifier>ISSN: 1344-6606</identifier><identifier>EISSN: 1881-3984</identifier><identifier>DOI: 10.3136/fstr.18.47</identifier><language>eng</language><publisher>Tsukuba: Japanese Society for Food Science and Technology</publisher><subject>detection sensitivity ; detection specificity ; hlyA gene ; Listeria monocytogenes ; real-time PCR</subject><ispartof>Food Science and Technology Research, 2012, Vol.18(1), pp.47-57</ispartof><rights>2012 by Japanese Society for Food Science and Technology</rights><rights>Copyright Japan Science and Technology Agency 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c613t-5d88b584afb9939581d43d96266b5f7bd7298629053b9f0d1beae85bf5571e913</citedby><cites>FETCH-LOGICAL-c613t-5d88b584afb9939581d43d96266b5f7bd7298629053b9f0d1beae85bf5571e913</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,4024,27923,27924,27925</link.rule.ids></links><search><creatorcontrib>LIU, Pei</creatorcontrib><creatorcontrib>MIZUE, Hiromi</creatorcontrib><creatorcontrib>FUJIHARA, Kumiko</creatorcontrib><creatorcontrib>KOBAYASHI, Hiroshi</creatorcontrib><creatorcontrib>KAMIKADO, Hideaki</creatorcontrib><creatorcontrib>TANAKA, Takashi</creatorcontrib><creatorcontrib>HONJOH, Ken-ichi</creatorcontrib><creatorcontrib>MIYAMOTO, Takahisa</creatorcontrib><creatorcontrib>Science and Technology Foundation</creatorcontrib><creatorcontrib>Kyushu University</creatorcontrib><creatorcontrib>Graduate School of Bioresource and Bioenvironmental Sciences</creatorcontrib><creatorcontrib>Department of Bioscience and Biotechnology</creatorcontrib><creatorcontrib>Research and Development Center</creatorcontrib><creatorcontrib>Faculty of Agriculture</creatorcontrib><creatorcontrib>Graduate School</creatorcontrib><creatorcontrib>Fukuoka Industry</creatorcontrib><creatorcontrib>Meiji Dairies Corporation</creatorcontrib><title>A New Rapid Real-Time PCR Method for Detection of Listeria monocytogenes Targeting the hlyA Gene</title><title>FOOD SCIENCE AND TECHNOLOGY RESEARCH</title><addtitle>Food Science and Technology Research</addtitle><description>In this study, primer sets of L. monocytogenes virulence gene hlyA were designed for the L. monocytogenes-specific PCR assay. 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In summary, this new PCR detection method is relatively sensitive and more specific to L. monocytogenes under certain conditions, could serve as a rapid detection method and has the potential for detection of L. monocytogenes from contaminated food.</description><subject>detection sensitivity</subject><subject>detection specificity</subject><subject>hlyA gene</subject><subject>Listeria monocytogenes</subject><subject>real-time PCR</subject><issn>1344-6606</issn><issn>1881-3984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><recordid>eNpdkU2rEzEUhgdR8Hp14y8IuBFhajL5XrgoVXuF-kGp65iZOWlTZiY1SZH--5syUsFFcsLJw_vmvKmq1wQvKKHivUs5LohaMPmkuiNKkZpqxZ6WM2WsFgKL59WLlI4YE65Vc1f9WqJv8Adt7cn3aAt2qHd-BPRjtUVfIR9Cj1yI6CNk6LIPEwoObXzKEL1FY5hCd8lhDxMktLNxD9lPe5QPgA7DZYnW5eJl9czZIcGrv_W--vn50271UG--r7-slpu6E4TmmvdKtVwx61qtqeaK9Iz2WjRCtNzJtpeNVqLRmNNWO9yTFiwo3jrOJQFN6H31dtY9xfD7DCmb0acOhsFOEM7JECJLHkxiXNA3_6HHcI5TeZ0hTFBVrBpRqHcz1cWQUgRnTtGPNl4MweYatrmGbYgyTBZ4PcMj9L6zQ5gGP8E_XTcyF0KfTINJY0r6CpNSZFnsunFZZtblv4rSh1npmLLdw83Uxuy7AW6mZHa-9buDjQYm-ghXQ52R</recordid><startdate>2012</startdate><enddate>2012</enddate><creator>LIU, Pei</creator><creator>MIZUE, Hiromi</creator><creator>FUJIHARA, Kumiko</creator><creator>KOBAYASHI, Hiroshi</creator><creator>KAMIKADO, Hideaki</creator><creator>TANAKA, Takashi</creator><creator>HONJOH, Ken-ichi</creator><creator>MIYAMOTO, Takahisa</creator><general>Japanese Society for Food Science and Technology</general><general>The Japanese Society for Food Science and Technology</general><general>Japan Science and Technology Agency</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7QR</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>K9.</scope><scope>P64</scope><scope>7QL</scope></search><sort><creationdate>2012</creationdate><title>A New Rapid Real-Time PCR Method for Detection of Listeria monocytogenes Targeting the hlyA Gene</title><author>LIU, Pei ; 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To evaluate the performance of these primer sets, the detection sensitivity and specificity were examined and the most suitable primer set was selected. Subsequently, it was subjected to compare the performance for detection of L. monocytogenes with commercially available kits (TaKaRa detection kit and ABI detection kit) on DNA level. Results of real-time PCR showed that the efficiency of this new primer set was 101.6% while TaKaRa and ABI detection kit were 101.1% and 107.4%, respectively. The detection sensitivity of all three methods was equivalent to less than 1 copy per reaction using purified genomic DNA as template. Detection specificity were 100% when testing L. monocytogenes isolates, and for other Listeria isolates were 15.4%, 76.9% and 100% by the method using hlyA L. monocytogenes detection primer set 7, TaKaRa and ABI detection kit, respectively. 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| subjects | detection sensitivity detection specificity hlyA gene Listeria monocytogenes real-time PCR |
| title | A New Rapid Real-Time PCR Method for Detection of Listeria monocytogenes Targeting the hlyA Gene |
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