Quantification and Discrimination of Viable and Dead Escherichia coli O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification
In this study, a rapid and sensitive method of real-time loop-mediated isothermal amplification (Rti-LAMP) assays was developed for quantification and discrimination of viable and heat-killed E. coli O157:H7 cells treated with low concentration of ethidium bromide monoazide (EMA). Four micrograms pe...
Gespeichert in:
| Veröffentlicht in: | Food analytical methods 2018-05, Vol.11 (5), p.1257-1266 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1266 |
|---|---|
| container_issue | 5 |
| container_start_page | 1257 |
| container_title | Food analytical methods |
| container_volume | 11 |
| creator | Zhao, Yuan Yang Tang, Kai Jie Zhang, Tian Tian Gao, Yan Yan Lin, Li Ping Wu, Guo Ping |
| description | In this study, a rapid and sensitive method of real-time loop-mediated isothermal amplification (Rti-LAMP) assays was developed for quantification and discrimination of viable and heat-killed
E. coli
O157:H7 cells treated with low concentration of ethidium bromide monoazide (EMA). Four micrograms per milliliter of EMA was chosen as the optimal concentration which did not inhibit DNA amplification derived from viable cells, but significantly increased the Tt values of dead cells in Rti-LAMP assays. When the DNA from 2.0 × 10
3
viable CFU of
E. coli
O157:H7 was subjected to EMA-Rti-LAMP, the resulting Tt value was 17.73 min. In contrast, the DNA from 2.0 × 10
3
CFU completely heat destroyed CFU of
E. coli
O157:H7 did not yield a positive amplification which Tt value was regarded as 60 min. When the DNA from viable plus heat-killed CFU at a ratio of 5:2995 was subjected to EMA-Rti-LAMP, the resulting Tt value was 23.06 min, which was statistically identical (
P
|
| doi_str_mv | 10.1007/s12161-017-1092-y |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_2021767591</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2021767591</sourcerecordid><originalsourceid>FETCH-LOGICAL-c316t-51ee8cde4651e9f7f8f09d0187cc74447a45c46e11232f66c5d85900236fe5623</originalsourceid><addsrcrecordid>eNp1UU1v1DAQjRBIlMIP4GaJs8HjJPaGW1kWWmmrCsTXzXLtMXFJ7MV2DsvP45fhVVA5cZqnmffmjeY1zXNgL4Ex-SoDBwGUgaTABk6PD5ozGFpBBym-PbzHPTxunuR8x5hgHfCz5veHRYfinTe6-BiIDpa89dkkP_uwtqIjX7y-nXAdorZkl82IyZvRa2Li5MkN9PL1pSRbnKZMXIoz2Y7e_MBAvvoyxqWQXTgJZgyF3B7Jroze-mUmbyrXWyTXMUT964Q-op5o8TOSfYwHeo3W64KWXOVYquusJ3IxH6b7m582j5yeMj77W8-bz-92n7aXdH_z_mp7saemBVFoD4gbY7ETFQ1Ouo1jg2WwkcbIruuk7nrTCQTgLXdCmN5u-oEx3gqHveDtefNi3XtI8eeCuai7uKRQLRVnHKSQ_QCVBSvLpJhzQqcO9Zc6HRUwdYpKrVGpGpU6RaWOVcNXTa7c8B3Tv83_F_0BSFKZVw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2021767591</pqid></control><display><type>article</type><title>Quantification and Discrimination of Viable and Dead Escherichia coli O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification</title><source>SpringerLink Journals - AutoHoldings</source><creator>Zhao, Yuan Yang ; Tang, Kai Jie ; Zhang, Tian Tian ; Gao, Yan Yan ; Lin, Li Ping ; Wu, Guo Ping</creator><creatorcontrib>Zhao, Yuan Yang ; Tang, Kai Jie ; Zhang, Tian Tian ; Gao, Yan Yan ; Lin, Li Ping ; Wu, Guo Ping</creatorcontrib><description>In this study, a rapid and sensitive method of real-time loop-mediated isothermal amplification (Rti-LAMP) assays was developed for quantification and discrimination of viable and heat-killed
E. coli
O157:H7 cells treated with low concentration of ethidium bromide monoazide (EMA). Four micrograms per milliliter of EMA was chosen as the optimal concentration which did not inhibit DNA amplification derived from viable cells, but significantly increased the Tt values of dead cells in Rti-LAMP assays. When the DNA from 2.0 × 10
3
viable CFU of
E. coli
O157:H7 was subjected to EMA-Rti-LAMP, the resulting Tt value was 17.73 min. In contrast, the DNA from 2.0 × 10
3
CFU completely heat destroyed CFU of
E. coli
O157:H7 did not yield a positive amplification which Tt value was regarded as 60 min. When the DNA from viable plus heat-killed CFU at a ratio of 5:2995 was subjected to EMA-Rti-LAMP, the resulting Tt value was 23.06 min, which was statistically identical (
P
< 0.05) to the Tt value of 24.07 min obtained with the DNA from only 5 viable CFU. The results indicate that even though 3.0 × 10
3
dead cells yielded a negative amplification setting the Tt value as 60 min, low numbers of viable cells in the presence of much higher numbers of dead cells still yielded a linear plot for enumerating viable CFU from Tt values. Detection of
E. coli
O157:H7 derived from contaminated chicken samples, the EMA-Rti-LAMP could notably distinguish viable and heat-killed cells from 5.0 × 10
1
to 1.0 × 10
4
CFU/g without enrichment.</description><identifier>ISSN: 1936-9751</identifier><identifier>EISSN: 1936-976X</identifier><identifier>DOI: 10.1007/s12161-017-1092-y</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Analytical Chemistry ; Chemistry ; Chemistry and Materials Science ; Chemistry/Food Science ; Deoxyribonucleic acid ; Discrimination ; DNA ; E coli ; Enrichment ; Ethidium bromide ; Food Science ; Heat ; Microbiology ; Real time ; Statistical analysis ; Statistical methods</subject><ispartof>Food analytical methods, 2018-05, Vol.11 (5), p.1257-1266</ispartof><rights>Springer Science+Business Media, LLC, part of Springer Nature 2017</rights><rights>Copyright Springer Science & Business Media 2018</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c316t-51ee8cde4651e9f7f8f09d0187cc74447a45c46e11232f66c5d85900236fe5623</citedby><cites>FETCH-LOGICAL-c316t-51ee8cde4651e9f7f8f09d0187cc74447a45c46e11232f66c5d85900236fe5623</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s12161-017-1092-y$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s12161-017-1092-y$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids></links><search><creatorcontrib>Zhao, Yuan Yang</creatorcontrib><creatorcontrib>Tang, Kai Jie</creatorcontrib><creatorcontrib>Zhang, Tian Tian</creatorcontrib><creatorcontrib>Gao, Yan Yan</creatorcontrib><creatorcontrib>Lin, Li Ping</creatorcontrib><creatorcontrib>Wu, Guo Ping</creatorcontrib><title>Quantification and Discrimination of Viable and Dead Escherichia coli O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification</title><title>Food analytical methods</title><addtitle>Food Anal. Methods</addtitle><description>In this study, a rapid and sensitive method of real-time loop-mediated isothermal amplification (Rti-LAMP) assays was developed for quantification and discrimination of viable and heat-killed
E. coli
O157:H7 cells treated with low concentration of ethidium bromide monoazide (EMA). Four micrograms per milliliter of EMA was chosen as the optimal concentration which did not inhibit DNA amplification derived from viable cells, but significantly increased the Tt values of dead cells in Rti-LAMP assays. When the DNA from 2.0 × 10
3
viable CFU of
E. coli
O157:H7 was subjected to EMA-Rti-LAMP, the resulting Tt value was 17.73 min. In contrast, the DNA from 2.0 × 10
3
CFU completely heat destroyed CFU of
E. coli
O157:H7 did not yield a positive amplification which Tt value was regarded as 60 min. When the DNA from viable plus heat-killed CFU at a ratio of 5:2995 was subjected to EMA-Rti-LAMP, the resulting Tt value was 23.06 min, which was statistically identical (
P
< 0.05) to the Tt value of 24.07 min obtained with the DNA from only 5 viable CFU. The results indicate that even though 3.0 × 10
3
dead cells yielded a negative amplification setting the Tt value as 60 min, low numbers of viable cells in the presence of much higher numbers of dead cells still yielded a linear plot for enumerating viable CFU from Tt values. Detection of
E. coli
O157:H7 derived from contaminated chicken samples, the EMA-Rti-LAMP could notably distinguish viable and heat-killed cells from 5.0 × 10
1
to 1.0 × 10
4
CFU/g without enrichment.</description><subject>Analytical Chemistry</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Chemistry/Food Science</subject><subject>Deoxyribonucleic acid</subject><subject>Discrimination</subject><subject>DNA</subject><subject>E coli</subject><subject>Enrichment</subject><subject>Ethidium bromide</subject><subject>Food Science</subject><subject>Heat</subject><subject>Microbiology</subject><subject>Real time</subject><subject>Statistical analysis</subject><subject>Statistical methods</subject><issn>1936-9751</issn><issn>1936-976X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNp1UU1v1DAQjRBIlMIP4GaJs8HjJPaGW1kWWmmrCsTXzXLtMXFJ7MV2DsvP45fhVVA5cZqnmffmjeY1zXNgL4Ex-SoDBwGUgaTABk6PD5ozGFpBBym-PbzHPTxunuR8x5hgHfCz5veHRYfinTe6-BiIDpa89dkkP_uwtqIjX7y-nXAdorZkl82IyZvRa2Li5MkN9PL1pSRbnKZMXIoz2Y7e_MBAvvoyxqWQXTgJZgyF3B7Jroze-mUmbyrXWyTXMUT964Q-op5o8TOSfYwHeo3W64KWXOVYquusJ3IxH6b7m582j5yeMj77W8-bz-92n7aXdH_z_mp7saemBVFoD4gbY7ETFQ1Ouo1jg2WwkcbIruuk7nrTCQTgLXdCmN5u-oEx3gqHveDtefNi3XtI8eeCuai7uKRQLRVnHKSQ_QCVBSvLpJhzQqcO9Zc6HRUwdYpKrVGpGpU6RaWOVcNXTa7c8B3Tv83_F_0BSFKZVw</recordid><startdate>20180501</startdate><enddate>20180501</enddate><creator>Zhao, Yuan Yang</creator><creator>Tang, Kai Jie</creator><creator>Zhang, Tian Tian</creator><creator>Gao, Yan Yan</creator><creator>Lin, Li Ping</creator><creator>Wu, Guo Ping</creator><general>Springer US</general><general>Springer Nature B.V</general><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>20180501</creationdate><title>Quantification and Discrimination of Viable and Dead Escherichia coli O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification</title><author>Zhao, Yuan Yang ; Tang, Kai Jie ; Zhang, Tian Tian ; Gao, Yan Yan ; Lin, Li Ping ; Wu, Guo Ping</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c316t-51ee8cde4651e9f7f8f09d0187cc74447a45c46e11232f66c5d85900236fe5623</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Analytical Chemistry</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Chemistry/Food Science</topic><topic>Deoxyribonucleic acid</topic><topic>Discrimination</topic><topic>DNA</topic><topic>E coli</topic><topic>Enrichment</topic><topic>Ethidium bromide</topic><topic>Food Science</topic><topic>Heat</topic><topic>Microbiology</topic><topic>Real time</topic><topic>Statistical analysis</topic><topic>Statistical methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhao, Yuan Yang</creatorcontrib><creatorcontrib>Tang, Kai Jie</creatorcontrib><creatorcontrib>Zhang, Tian Tian</creatorcontrib><creatorcontrib>Gao, Yan Yan</creatorcontrib><creatorcontrib>Lin, Li Ping</creatorcontrib><creatorcontrib>Wu, Guo Ping</creatorcontrib><collection>CrossRef</collection><jtitle>Food analytical methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, Yuan Yang</au><au>Tang, Kai Jie</au><au>Zhang, Tian Tian</au><au>Gao, Yan Yan</au><au>Lin, Li Ping</au><au>Wu, Guo Ping</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification and Discrimination of Viable and Dead Escherichia coli O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification</atitle><jtitle>Food analytical methods</jtitle><stitle>Food Anal. Methods</stitle><date>2018-05-01</date><risdate>2018</risdate><volume>11</volume><issue>5</issue><spage>1257</spage><epage>1266</epage><pages>1257-1266</pages><issn>1936-9751</issn><eissn>1936-976X</eissn><abstract>In this study, a rapid and sensitive method of real-time loop-mediated isothermal amplification (Rti-LAMP) assays was developed for quantification and discrimination of viable and heat-killed
E. coli
O157:H7 cells treated with low concentration of ethidium bromide monoazide (EMA). Four micrograms per milliliter of EMA was chosen as the optimal concentration which did not inhibit DNA amplification derived from viable cells, but significantly increased the Tt values of dead cells in Rti-LAMP assays. When the DNA from 2.0 × 10
3
viable CFU of
E. coli
O157:H7 was subjected to EMA-Rti-LAMP, the resulting Tt value was 17.73 min. In contrast, the DNA from 2.0 × 10
3
CFU completely heat destroyed CFU of
E. coli
O157:H7 did not yield a positive amplification which Tt value was regarded as 60 min. When the DNA from viable plus heat-killed CFU at a ratio of 5:2995 was subjected to EMA-Rti-LAMP, the resulting Tt value was 23.06 min, which was statistically identical (
P
< 0.05) to the Tt value of 24.07 min obtained with the DNA from only 5 viable CFU. The results indicate that even though 3.0 × 10
3
dead cells yielded a negative amplification setting the Tt value as 60 min, low numbers of viable cells in the presence of much higher numbers of dead cells still yielded a linear plot for enumerating viable CFU from Tt values. Detection of
E. coli
O157:H7 derived from contaminated chicken samples, the EMA-Rti-LAMP could notably distinguish viable and heat-killed cells from 5.0 × 10
1
to 1.0 × 10
4
CFU/g without enrichment.</abstract><cop>New York</cop><pub>Springer US</pub><doi>10.1007/s12161-017-1092-y</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1936-9751 |
| ispartof | Food analytical methods, 2018-05, Vol.11 (5), p.1257-1266 |
| issn | 1936-9751 1936-976X |
| language | eng |
| recordid | cdi_proquest_journals_2021767591 |
| source | SpringerLink Journals - AutoHoldings |
| subjects | Analytical Chemistry Chemistry Chemistry and Materials Science Chemistry/Food Science Deoxyribonucleic acid Discrimination DNA E coli Enrichment Ethidium bromide Food Science Heat Microbiology Real time Statistical analysis Statistical methods |
| title | Quantification and Discrimination of Viable and Dead Escherichia coli O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-08T02%3A51%3A54IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Quantification%20and%20Discrimination%20of%20Viable%20and%20Dead%20Escherichia%20coli%20O157:H7%20Cells%20from%20Chicken%20Without%20Enrichment%20by%20Ethidium%20Bromide%20Monoazide%20Real-time%20Loop-Mediated%20Isothermal%20Amplification&rft.jtitle=Food%20analytical%20methods&rft.au=Zhao,%20Yuan%20Yang&rft.date=2018-05-01&rft.volume=11&rft.issue=5&rft.spage=1257&rft.epage=1266&rft.pages=1257-1266&rft.issn=1936-9751&rft.eissn=1936-976X&rft_id=info:doi/10.1007/s12161-017-1092-y&rft_dat=%3Cproquest_cross%3E2021767591%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2021767591&rft_id=info:pmid/&rfr_iscdi=true |