Development of a surface plasmon resonance-based immunoassay for Listeria monocytogenes
A polyclonal antibody was produced against Internalin B (InlB)-enriched extract and used to develop an inhibition assay to detect Listeria monocytogenes cells in solution using surface plasmon resonance. The gene sequence encoding for the InlB protein was cloned into a Qiagen pQE-60 vector, expresse...
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| Veröffentlicht in: | Journal of food protection 2005-04, Vol.68 (4), p.728-735 |
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| container_title | Journal of food protection |
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| creator | Leonard, P Hearty, S Wyatt, G Quinn, J O'Kennedy, R |
| description | A polyclonal antibody was produced against Internalin B (InlB)-enriched extract and used to develop an inhibition assay to detect Listeria monocytogenes cells in solution using surface plasmon resonance. The gene sequence encoding for the InlB protein was cloned into a Qiagen pQE-60 vector, expressed in Escherichia coli, and purified by immobilized metal affinity chromatography. Protein G-purified anti-InlB-enriched extract polyclonal antibody was incubated with various concentrations of L. monocytogenes cells and subsequently injected over a purified-recombinant InlB (rInlB)-immobilized CM5 sensor chip surface. A decrease in antibody binding response was observed with increasing L. monocytogenes cell concentrations. Intraday and interday assay variability studies were carried out to evaluate precision and reproducibility. The assay had a limit of detection of less than 2 x 10(5) cells per ml and could be successfully reproduced with coefficients of variation of between 2.5 and 7.7%. |
| doi_str_mv | 10.4315/0362-028X-68.4.728 |
| format | Article |
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The gene sequence encoding for the InlB protein was cloned into a Qiagen pQE-60 vector, expressed in Escherichia coli, and purified by immobilized metal affinity chromatography. Protein G-purified anti-InlB-enriched extract polyclonal antibody was incubated with various concentrations of L. monocytogenes cells and subsequently injected over a purified-recombinant InlB (rInlB)-immobilized CM5 sensor chip surface. A decrease in antibody binding response was observed with increasing L. monocytogenes cell concentrations. Intraday and interday assay variability studies were carried out to evaluate precision and reproducibility. The assay had a limit of detection of less than 2 x 10(5) cells per ml and could be successfully reproduced with coefficients of variation of between 2.5 and 7.7%.</description><identifier>ISSN: 0362-028X</identifier><identifier>EISSN: 1944-9097</identifier><identifier>DOI: 10.4315/0362-028X-68.4.728</identifier><identifier>PMID: 15830663</identifier><identifier>CODEN: JFPRDR</identifier><language>eng</language><publisher>Des Moines, IA: International Association of Milk, Food and Environmental Sanitarians</publisher><subject>Animals ; Antibodies ; bacterial contamination ; bacterial proteins ; Biological and medical sciences ; DNA, Bacterial - analysis ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Fish and seafood industries ; food contamination ; Food Contamination - analysis ; Food industries ; Food Microbiology ; food pathogens ; Fundamental and applied biological sciences. Psychology ; genes ; HACCP ; immunoassays ; Listeria monocytogenes ; Listeria monocytogenes - immunology ; Listeria monocytogenes - isolation & purification ; methodology ; microbial detection ; Molecular Weight ; polyclonal antibodies ; quality control ; Reproducibility of Results ; Sensitivity and Specificity ; Surface Plasmon Resonance - methods ; virulence</subject><ispartof>Journal of food protection, 2005-04, Vol.68 (4), p.728-735</ispartof><rights>2005 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c430t-a3dbf62ef9fb75e2227beafcfdc05f2ca11231bf9feefce37c95a248a74947cd3</citedby><cites>FETCH-LOGICAL-c430t-a3dbf62ef9fb75e2227beafcfdc05f2ca11231bf9feefce37c95a248a74947cd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=16680497$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15830663$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Leonard, P</creatorcontrib><creatorcontrib>Hearty, S</creatorcontrib><creatorcontrib>Wyatt, G</creatorcontrib><creatorcontrib>Quinn, J</creatorcontrib><creatorcontrib>O'Kennedy, R</creatorcontrib><title>Development of a surface plasmon resonance-based immunoassay for Listeria monocytogenes</title><title>Journal of food protection</title><addtitle>J Food Prot</addtitle><description>A polyclonal antibody was produced against Internalin B (InlB)-enriched extract and used to develop an inhibition assay to detect Listeria monocytogenes cells in solution using surface plasmon resonance. The gene sequence encoding for the InlB protein was cloned into a Qiagen pQE-60 vector, expressed in Escherichia coli, and purified by immobilized metal affinity chromatography. Protein G-purified anti-InlB-enriched extract polyclonal antibody was incubated with various concentrations of L. monocytogenes cells and subsequently injected over a purified-recombinant InlB (rInlB)-immobilized CM5 sensor chip surface. A decrease in antibody binding response was observed with increasing L. monocytogenes cell concentrations. Intraday and interday assay variability studies were carried out to evaluate precision and reproducibility. The assay had a limit of detection of less than 2 x 10(5) cells per ml and could be successfully reproduced with coefficients of variation of between 2.5 and 7.7%.</description><subject>Animals</subject><subject>Antibodies</subject><subject>bacterial contamination</subject><subject>bacterial proteins</subject><subject>Biological and medical sciences</subject><subject>DNA, Bacterial - analysis</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Escherichia coli</subject><subject>Fish and seafood industries</subject><subject>food contamination</subject><subject>Food Contamination - analysis</subject><subject>Food industries</subject><subject>Food Microbiology</subject><subject>food pathogens</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>genes</subject><subject>HACCP</subject><subject>immunoassays</subject><subject>Listeria monocytogenes</subject><subject>Listeria monocytogenes - immunology</subject><subject>Listeria monocytogenes - isolation & purification</subject><subject>methodology</subject><subject>microbial detection</subject><subject>Molecular Weight</subject><subject>polyclonal antibodies</subject><subject>quality control</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Surface Plasmon Resonance - methods</subject><subject>virulence</subject><issn>0362-028X</issn><issn>1944-9097</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpF0E1v1DAQgGELgei29A9wAF_oLYu_EsdHVD6llThA1d6siTOugpJ48SRI--_r1a7oyYd5ZiS_jL2VYmu0rD8K3ahKqPahatqt2VrVvmAb6YypnHD2Jdv8BxfskuiPEEI51bxmF7JutWgavWH3n_Efjmk_4bzwFDlwWnOEgHw_Ak1p5hkpzTAHrDog7PkwTeucgAgOPKbMdwMtmAfgBadwWNIjzkhv2KsII-H1-b1id1-__L79Xu1-fvtx-2lXBaPFUoHuu9gojC52tkallO0QYoh9EHVUAaRUWnZljBgDahtcDcq0YI0zNvT6it2c7u5z-rsiLX4aKOA4woxpJS-taWujRIHqBENORBmj3-dhgnzwUvhjTn-s5Y-1fNN640vOsvTufH3tJuyfV879CvhwBkABxphLqIGeXdO0wjhb3PuTi5A8POZi7n4pIbUQzpVfK_0EizOJUQ</recordid><startdate>20050401</startdate><enddate>20050401</enddate><creator>Leonard, P</creator><creator>Hearty, S</creator><creator>Wyatt, G</creator><creator>Quinn, J</creator><creator>O'Kennedy, R</creator><general>International Association of Milk, Food and Environmental Sanitarians</general><scope>FBQ</scope><scope>IQODW</scope><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>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20050401</creationdate><title>Development of a surface plasmon resonance-based immunoassay for Listeria monocytogenes</title><author>Leonard, P ; Hearty, S ; Wyatt, G ; Quinn, J ; O'Kennedy, R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c430t-a3dbf62ef9fb75e2227beafcfdc05f2ca11231bf9feefce37c95a248a74947cd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>bacterial contamination</topic><topic>bacterial proteins</topic><topic>Biological and medical sciences</topic><topic>DNA, Bacterial - analysis</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Escherichia coli</topic><topic>Fish and seafood industries</topic><topic>food contamination</topic><topic>Food Contamination - analysis</topic><topic>Food industries</topic><topic>Food Microbiology</topic><topic>food pathogens</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>genes</topic><topic>HACCP</topic><topic>immunoassays</topic><topic>Listeria monocytogenes</topic><topic>Listeria monocytogenes - immunology</topic><topic>Listeria monocytogenes - isolation & purification</topic><topic>methodology</topic><topic>microbial detection</topic><topic>Molecular Weight</topic><topic>polyclonal antibodies</topic><topic>quality control</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Surface Plasmon Resonance - methods</topic><topic>virulence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Leonard, P</creatorcontrib><creatorcontrib>Hearty, S</creatorcontrib><creatorcontrib>Wyatt, G</creatorcontrib><creatorcontrib>Quinn, J</creatorcontrib><creatorcontrib>O'Kennedy, R</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of food protection</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Leonard, P</au><au>Hearty, S</au><au>Wyatt, G</au><au>Quinn, J</au><au>O'Kennedy, R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a surface plasmon resonance-based immunoassay for Listeria monocytogenes</atitle><jtitle>Journal of food protection</jtitle><addtitle>J Food Prot</addtitle><date>2005-04-01</date><risdate>2005</risdate><volume>68</volume><issue>4</issue><spage>728</spage><epage>735</epage><pages>728-735</pages><issn>0362-028X</issn><eissn>1944-9097</eissn><coden>JFPRDR</coden><abstract>A polyclonal antibody was produced against Internalin B (InlB)-enriched extract and used to develop an inhibition assay to detect Listeria monocytogenes cells in solution using surface plasmon resonance. The gene sequence encoding for the InlB protein was cloned into a Qiagen pQE-60 vector, expressed in Escherichia coli, and purified by immobilized metal affinity chromatography. Protein G-purified anti-InlB-enriched extract polyclonal antibody was incubated with various concentrations of L. monocytogenes cells and subsequently injected over a purified-recombinant InlB (rInlB)-immobilized CM5 sensor chip surface. A decrease in antibody binding response was observed with increasing L. monocytogenes cell concentrations. Intraday and interday assay variability studies were carried out to evaluate precision and reproducibility. The assay had a limit of detection of less than 2 x 10(5) cells per ml and could be successfully reproduced with coefficients of variation of between 2.5 and 7.7%.</abstract><cop>Des Moines, IA</cop><pub>International Association of Milk, Food and Environmental Sanitarians</pub><pmid>15830663</pmid><doi>10.4315/0362-028X-68.4.728</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Antibodies bacterial contamination bacterial proteins Biological and medical sciences DNA, Bacterial - analysis Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Escherichia coli Fish and seafood industries food contamination Food Contamination - analysis Food industries Food Microbiology food pathogens Fundamental and applied biological sciences. Psychology genes HACCP immunoassays Listeria monocytogenes Listeria monocytogenes - immunology Listeria monocytogenes - isolation & purification methodology microbial detection Molecular Weight polyclonal antibodies quality control Reproducibility of Results Sensitivity and Specificity Surface Plasmon Resonance - methods virulence |
| title | Development of a surface plasmon resonance-based immunoassay for Listeria monocytogenes |
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