Immunodetection of Streptococcus uberis pathogen in raw milk
•Anti - Str. uberis antibodies characterized experimentally and in silico.•Immunobiosensor for rapid selective detection of pathogen Str. Uberis.•Pathogen identification in mastitis milk in agreement with microbiological plating. Streptococcus uberis is a major mastitis-causing environmental pathoge...
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| Veröffentlicht in: | Enzyme and microbial technology 2019-11, Vol.130, p.109360-109360, Article 109360 |
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| creator | Mihklepp, K. Kivirand, K. Juronen, D. Lõokene, A. Rinken, T. |
| description | •Anti - Str. uberis antibodies characterized experimentally and in silico.•Immunobiosensor for rapid selective detection of pathogen Str. Uberis.•Pathogen identification in mastitis milk in agreement with microbiological plating.
Streptococcus uberis is a major mastitis-causing environmental pathogen, which rapid immunodetection has not been possible due to the absence of specific anti-Str. uberis antibodies. Recently, a specific antibody against the Str. uberis adhesion molecule (SUAM) has been designed. In the present study, the specificity and affinity of this antibody towards SUAM antigenic region SAPVYLGVSTE and Str. uberis cells are characterized, using experimental and in silico bioinformatic methods. The selectivity studies and bioinformatic analyses revealed high specificity of the antibody towards Str. uberis. The Kd value of SAPVYLGVSTE/anti-Str. uberis antibody complex was 27 ± 6 nM, indicating the applicability of this antibody for the detection of Str. uberis. The anti-Str. uberis antibody was used as a specific biorecognition element of a biosensor for the detection of Str. uberis bacteria in phosphate buffer and in milk and these analyses took less than 20 min. The Str. uberis biosensor was also tested in the milk of cows suffering from mastitis and the obtained results were in good agreement with the conventional identification of this pathogen by microbiological plating. |
| doi_str_mv | 10.1016/j.enzmictec.2019.109360 |
| format | Article |
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Streptococcus uberis is a major mastitis-causing environmental pathogen, which rapid immunodetection has not been possible due to the absence of specific anti-Str. uberis antibodies. Recently, a specific antibody against the Str. uberis adhesion molecule (SUAM) has been designed. In the present study, the specificity and affinity of this antibody towards SUAM antigenic region SAPVYLGVSTE and Str. uberis cells are characterized, using experimental and in silico bioinformatic methods. The selectivity studies and bioinformatic analyses revealed high specificity of the antibody towards Str. uberis. The Kd value of SAPVYLGVSTE/anti-Str. uberis antibody complex was 27 ± 6 nM, indicating the applicability of this antibody for the detection of Str. uberis. The anti-Str. uberis antibody was used as a specific biorecognition element of a biosensor for the detection of Str. uberis bacteria in phosphate buffer and in milk and these analyses took less than 20 min. The Str. uberis biosensor was also tested in the milk of cows suffering from mastitis and the obtained results were in good agreement with the conventional identification of this pathogen by microbiological plating.</description><identifier>ISSN: 0141-0229</identifier><identifier>EISSN: 1879-0909</identifier><identifier>DOI: 10.1016/j.enzmictec.2019.109360</identifier><identifier>PMID: 31421723</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Affinity ; Animals ; Anti-Str. uberis antibodies ; Antibodies, Bacterial - blood ; Antibody Affinity ; Bacterial Adhesion ; Bioinformatic analysis ; Biosensing Techniques ; Cattle ; Computational Biology ; Female ; Immunodetection of Str. uberis ; Mastitis, Bovine - diagnosis ; Mastitis, Bovine - immunology ; Milk - microbiology ; Milk analysis ; Raw Foods - microbiology ; Streptococcal Infections - diagnosis ; Streptococcal Infections - immunology ; Streptococcal Infections - veterinary ; Streptococcus - isolation & purification ; Streptococcus - pathogenicity</subject><ispartof>Enzyme and microbial technology, 2019-11, Vol.130, p.109360-109360, Article 109360</ispartof><rights>2019 Elsevier Inc.</rights><rights>Copyright © 2019 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c457t-b77d6528183a42d61c1d685f28c2a0ffc1c68cfd9f542171f0a749a84b1732293</citedby><cites>FETCH-LOGICAL-c457t-b77d6528183a42d61c1d685f28c2a0ffc1c68cfd9f542171f0a749a84b1732293</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.enzmictec.2019.109360$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31421723$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mihklepp, K.</creatorcontrib><creatorcontrib>Kivirand, K.</creatorcontrib><creatorcontrib>Juronen, D.</creatorcontrib><creatorcontrib>Lõokene, A.</creatorcontrib><creatorcontrib>Rinken, T.</creatorcontrib><title>Immunodetection of Streptococcus uberis pathogen in raw milk</title><title>Enzyme and microbial technology</title><addtitle>Enzyme Microb Technol</addtitle><description>•Anti - Str. uberis antibodies characterized experimentally and in silico.•Immunobiosensor for rapid selective detection of pathogen Str. Uberis.•Pathogen identification in mastitis milk in agreement with microbiological plating.
Streptococcus uberis is a major mastitis-causing environmental pathogen, which rapid immunodetection has not been possible due to the absence of specific anti-Str. uberis antibodies. Recently, a specific antibody against the Str. uberis adhesion molecule (SUAM) has been designed. In the present study, the specificity and affinity of this antibody towards SUAM antigenic region SAPVYLGVSTE and Str. uberis cells are characterized, using experimental and in silico bioinformatic methods. The selectivity studies and bioinformatic analyses revealed high specificity of the antibody towards Str. uberis. The Kd value of SAPVYLGVSTE/anti-Str. uberis antibody complex was 27 ± 6 nM, indicating the applicability of this antibody for the detection of Str. uberis. The anti-Str. uberis antibody was used as a specific biorecognition element of a biosensor for the detection of Str. uberis bacteria in phosphate buffer and in milk and these analyses took less than 20 min. The Str. uberis biosensor was also tested in the milk of cows suffering from mastitis and the obtained results were in good agreement with the conventional identification of this pathogen by microbiological plating.</description><subject>Affinity</subject><subject>Animals</subject><subject>Anti-Str. uberis antibodies</subject><subject>Antibodies, Bacterial - blood</subject><subject>Antibody Affinity</subject><subject>Bacterial Adhesion</subject><subject>Bioinformatic analysis</subject><subject>Biosensing Techniques</subject><subject>Cattle</subject><subject>Computational Biology</subject><subject>Female</subject><subject>Immunodetection of Str. uberis</subject><subject>Mastitis, Bovine - diagnosis</subject><subject>Mastitis, Bovine - immunology</subject><subject>Milk - microbiology</subject><subject>Milk analysis</subject><subject>Raw Foods - microbiology</subject><subject>Streptococcal Infections - diagnosis</subject><subject>Streptococcal Infections - immunology</subject><subject>Streptococcal Infections - veterinary</subject><subject>Streptococcus - isolation & purification</subject><subject>Streptococcus - pathogenicity</subject><issn>0141-0229</issn><issn>1879-0909</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtOwzAQRS0EoqXwC5AlmxSP83AssakQj0qVWABry_EDXJK42AkIvh5XKd2iWYw0unPvzEHoAvAcMJRX67nuflorey3nBAOLU5aV-ABNoaIsxQyzQzTFkEOKCWETdBLCGuM4yPExmmSQE6Akm6LrZdsOnVM6OvXWdYkzyVPv9aZ30kk5hGSotbch2Yj-zb3qLrFd4sVX0trm_RQdGdEEfbbrM_Ryd_t885CuHu-XN4tVKvOC9mlNqSoLUkGViZyoEiSosioMqSQR2BgJsqykUcwU27PAYEFzJqq8BprF67MZuhx9N959DDr0vLVB6qYRnXZD4ITQgsXKaJTSUSq9C8FrwzfetsJ_c8B8i46v-R4d36LjI7q4eb4LGepWq_3eH6soWIwCHV_9tNrzIK3upFbWR3hcOftvyC-z84Mq</recordid><startdate>20191101</startdate><enddate>20191101</enddate><creator>Mihklepp, K.</creator><creator>Kivirand, K.</creator><creator>Juronen, D.</creator><creator>Lõokene, A.</creator><creator>Rinken, T.</creator><general>Elsevier Inc</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>20191101</creationdate><title>Immunodetection of Streptococcus uberis pathogen in raw milk</title><author>Mihklepp, K. ; Kivirand, K. ; Juronen, D. ; Lõokene, A. ; Rinken, T.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c457t-b77d6528183a42d61c1d685f28c2a0ffc1c68cfd9f542171f0a749a84b1732293</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Affinity</topic><topic>Animals</topic><topic>Anti-Str. uberis antibodies</topic><topic>Antibodies, Bacterial - blood</topic><topic>Antibody Affinity</topic><topic>Bacterial Adhesion</topic><topic>Bioinformatic analysis</topic><topic>Biosensing Techniques</topic><topic>Cattle</topic><topic>Computational Biology</topic><topic>Female</topic><topic>Immunodetection of Str. uberis</topic><topic>Mastitis, Bovine - diagnosis</topic><topic>Mastitis, Bovine - immunology</topic><topic>Milk - microbiology</topic><topic>Milk analysis</topic><topic>Raw Foods - microbiology</topic><topic>Streptococcal Infections - diagnosis</topic><topic>Streptococcal Infections - immunology</topic><topic>Streptococcal Infections - veterinary</topic><topic>Streptococcus - isolation & purification</topic><topic>Streptococcus - pathogenicity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mihklepp, K.</creatorcontrib><creatorcontrib>Kivirand, K.</creatorcontrib><creatorcontrib>Juronen, D.</creatorcontrib><creatorcontrib>Lõokene, A.</creatorcontrib><creatorcontrib>Rinken, T.</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>Enzyme and microbial technology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mihklepp, K.</au><au>Kivirand, K.</au><au>Juronen, D.</au><au>Lõokene, A.</au><au>Rinken, T.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Immunodetection of Streptococcus uberis pathogen in raw milk</atitle><jtitle>Enzyme and microbial technology</jtitle><addtitle>Enzyme Microb Technol</addtitle><date>2019-11-01</date><risdate>2019</risdate><volume>130</volume><spage>109360</spage><epage>109360</epage><pages>109360-109360</pages><artnum>109360</artnum><issn>0141-0229</issn><eissn>1879-0909</eissn><abstract>•Anti - Str. uberis antibodies characterized experimentally and in silico.•Immunobiosensor for rapid selective detection of pathogen Str. Uberis.•Pathogen identification in mastitis milk in agreement with microbiological plating.
Streptococcus uberis is a major mastitis-causing environmental pathogen, which rapid immunodetection has not been possible due to the absence of specific anti-Str. uberis antibodies. Recently, a specific antibody against the Str. uberis adhesion molecule (SUAM) has been designed. In the present study, the specificity and affinity of this antibody towards SUAM antigenic region SAPVYLGVSTE and Str. uberis cells are characterized, using experimental and in silico bioinformatic methods. The selectivity studies and bioinformatic analyses revealed high specificity of the antibody towards Str. uberis. The Kd value of SAPVYLGVSTE/anti-Str. uberis antibody complex was 27 ± 6 nM, indicating the applicability of this antibody for the detection of Str. uberis. The anti-Str. uberis antibody was used as a specific biorecognition element of a biosensor for the detection of Str. uberis bacteria in phosphate buffer and in milk and these analyses took less than 20 min. The Str. uberis biosensor was also tested in the milk of cows suffering from mastitis and the obtained results were in good agreement with the conventional identification of this pathogen by microbiological plating.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>31421723</pmid><doi>10.1016/j.enzmictec.2019.109360</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Affinity Animals Anti-Str. uberis antibodies Antibodies, Bacterial - blood Antibody Affinity Bacterial Adhesion Bioinformatic analysis Biosensing Techniques Cattle Computational Biology Female Immunodetection of Str. uberis Mastitis, Bovine - diagnosis Mastitis, Bovine - immunology Milk - microbiology Milk analysis Raw Foods - microbiology Streptococcal Infections - diagnosis Streptococcal Infections - immunology Streptococcal Infections - veterinary Streptococcus - isolation & purification Streptococcus - pathogenicity |
| title | Immunodetection of Streptococcus uberis pathogen in raw milk |
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