Comparative evaluation of recombinant LigB protein and heat-killed antigen-based latex agglutination test with microscopic agglutination test for diagnosis of bovine leptospirosis
This study aimed to develop latex agglutination test (LAT) using recombinant leptospiral immunoglobulin-like protein (LigB) (rLigB) antigen and compare its diagnostic efficacy with LAT using conventional heat-killed leptospiral antigen and microscopic agglutination test (MAT) in diagnosing bovine le...
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| Veröffentlicht in: | Tropical animal health and production 2015-10, Vol.47 (7), p.1329-1335 |
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| creator | Nagalingam, Mohandoss Thirumalesh, Sushma Rahim Assadi Kalleshamurthy, Triveni Niharika, Nakkala Balamurugan, Vinayagamurthy Shome, Rajeswari Sengupta, Pinaki Prasad Shome, Bibek Ranjan Prabhudas, Krishnamsetty Rahman, Habibur |
| description | This study aimed to develop latex agglutination test (LAT) using recombinant leptospiral immunoglobulin-like protein (LigB) (rLigB) antigen and compare its diagnostic efficacy with LAT using conventional heat-killed leptospiral antigen and microscopic agglutination test (MAT) in diagnosing bovine leptospirosis. The PCR-amplified 1053-bp ligB gene sequences from Leptospira borgpetersenii Hardjo serovar were cloned in pET 32 (a) vector at EcoRI and NotI sites and expressed in BL21 E. coli cells as fusion protein with thioredoxin (−57 kDa) and characterized by SDS-PAGE and immunoblot. Out of 390 serum samples [cattle (n = 214), buffaloes (n = 176)] subjected to MAT, 115 samples showed reciprocal titre≥100 up to 1600 against one or more serovars. For recombinant LigB protein/antigen-based LAT, agglutination was observed in the positive sample, while no agglutination was observed in the negative sample. Similarly, heat-killed leptospiral antigen was prepared from and used in LAT for comparison with MAT. A two-sided contingency table was used for analysis of LAT using both the antigens separately against MAT for 390 serum samples. The sensitivity, specificity and positive and negative predictive values of recombinant LigB LAT were found to be 75.65, 91.27, 78.38 and 89.96 %, respectively, and that of heat-killed antigen-based LAT were 72.17, 89.82, 74.77 and 88.53 %, respectively, in comparison with MAT. This developed test will be an alternative/complementary to the existing battery of diagnostic assays/tests for specific detection of pathogenic Leptospira infection in bovine population. |
| doi_str_mv | 10.1007/s11250-015-0867-7 |
| format | Article |
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The PCR-amplified 1053-bp ligB gene sequences from Leptospira borgpetersenii Hardjo serovar were cloned in pET 32 (a) vector at EcoRI and NotI sites and expressed in BL21 E. coli cells as fusion protein with thioredoxin (−57 kDa) and characterized by SDS-PAGE and immunoblot. Out of 390 serum samples [cattle (n = 214), buffaloes (n = 176)] subjected to MAT, 115 samples showed reciprocal titre≥100 up to 1600 against one or more serovars. For recombinant LigB protein/antigen-based LAT, agglutination was observed in the positive sample, while no agglutination was observed in the negative sample. Similarly, heat-killed leptospiral antigen was prepared from and used in LAT for comparison with MAT. A two-sided contingency table was used for analysis of LAT using both the antigens separately against MAT for 390 serum samples. The sensitivity, specificity and positive and negative predictive values of recombinant LigB LAT were found to be 75.65, 91.27, 78.38 and 89.96 %, respectively, and that of heat-killed antigen-based LAT were 72.17, 89.82, 74.77 and 88.53 %, respectively, in comparison with MAT. This developed test will be an alternative/complementary to the existing battery of diagnostic assays/tests for specific detection of pathogenic Leptospira infection in bovine population.</description><identifier>ISSN: 0049-4747</identifier><identifier>EISSN: 1573-7438</identifier><identifier>DOI: 10.1007/s11250-015-0867-7</identifier><identifier>PMID: 26065562</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Animals ; Antibodies, Bacterial - immunology ; Antigens, Bacterial - immunology ; Biomedical and Life Sciences ; Cattle ; Cattle Diseases - blood ; Cattle Diseases - diagnosis ; Cattle Diseases - immunology ; E coli ; Electrophoresis, Polyacrylamide Gel - veterinary ; Enzyme-Linked Immunosorbent Assay - veterinary ; Escherichia coli ; Latex Fixation Tests - veterinary ; Leptospira ; Leptospira - immunology ; Leptospira borgpetersenii ; Leptospirosis - diagnosis ; Leptospirosis - immunology ; Leptospirosis - veterinary ; Life Sciences ; Recombinant Proteins - immunology ; Regular Articles ; Sensitivity and Specificity ; Serologic Tests - veterinary ; Veterinary Medicine/Veterinary Science ; Zoology</subject><ispartof>Tropical animal health and production, 2015-10, Vol.47 (7), p.1329-1335</ispartof><rights>Springer Science+Business Media Dordrecht 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c542t-57c8f3a1122f7c9718dcf13fbc7d1c79e21f041e7021826fedcb214308b4f0553</citedby><cites>FETCH-LOGICAL-c542t-57c8f3a1122f7c9718dcf13fbc7d1c79e21f041e7021826fedcb214308b4f0553</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/s11250-015-0867-7$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11250-015-0867-7$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27923,27924,41487,42556,51318</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26065562$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nagalingam, Mohandoss</creatorcontrib><creatorcontrib>Thirumalesh, Sushma Rahim Assadi</creatorcontrib><creatorcontrib>Kalleshamurthy, Triveni</creatorcontrib><creatorcontrib>Niharika, Nakkala</creatorcontrib><creatorcontrib>Balamurugan, Vinayagamurthy</creatorcontrib><creatorcontrib>Shome, Rajeswari</creatorcontrib><creatorcontrib>Sengupta, Pinaki Prasad</creatorcontrib><creatorcontrib>Shome, Bibek Ranjan</creatorcontrib><creatorcontrib>Prabhudas, Krishnamsetty</creatorcontrib><creatorcontrib>Rahman, Habibur</creatorcontrib><title>Comparative evaluation of recombinant LigB protein and heat-killed antigen-based latex agglutination test with microscopic agglutination test for diagnosis of bovine leptospirosis</title><title>Tropical animal health and production</title><addtitle>Trop Anim Health Prod</addtitle><addtitle>Trop Anim Health Prod</addtitle><description>This study aimed to develop latex agglutination test (LAT) using recombinant leptospiral immunoglobulin-like protein (LigB) (rLigB) antigen and compare its diagnostic efficacy with LAT using conventional heat-killed leptospiral antigen and microscopic agglutination test (MAT) in diagnosing bovine leptospirosis. The PCR-amplified 1053-bp ligB gene sequences from Leptospira borgpetersenii Hardjo serovar were cloned in pET 32 (a) vector at EcoRI and NotI sites and expressed in BL21 E. coli cells as fusion protein with thioredoxin (−57 kDa) and characterized by SDS-PAGE and immunoblot. Out of 390 serum samples [cattle (n = 214), buffaloes (n = 176)] subjected to MAT, 115 samples showed reciprocal titre≥100 up to 1600 against one or more serovars. For recombinant LigB protein/antigen-based LAT, agglutination was observed in the positive sample, while no agglutination was observed in the negative sample. Similarly, heat-killed leptospiral antigen was prepared from and used in LAT for comparison with MAT. A two-sided contingency table was used for analysis of LAT using both the antigens separately against MAT for 390 serum samples. The sensitivity, specificity and positive and negative predictive values of recombinant LigB LAT were found to be 75.65, 91.27, 78.38 and 89.96 %, respectively, and that of heat-killed antigen-based LAT were 72.17, 89.82, 74.77 and 88.53 %, respectively, in comparison with MAT. This developed test will be an alternative/complementary to the existing battery of diagnostic assays/tests for specific detection of pathogenic Leptospira infection in bovine population.</description><subject>Animals</subject><subject>Antibodies, Bacterial - immunology</subject><subject>Antigens, Bacterial - immunology</subject><subject>Biomedical and Life Sciences</subject><subject>Cattle</subject><subject>Cattle Diseases - blood</subject><subject>Cattle Diseases - diagnosis</subject><subject>Cattle Diseases - immunology</subject><subject>E coli</subject><subject>Electrophoresis, Polyacrylamide Gel - veterinary</subject><subject>Enzyme-Linked Immunosorbent Assay - veterinary</subject><subject>Escherichia coli</subject><subject>Latex Fixation Tests - veterinary</subject><subject>Leptospira</subject><subject>Leptospira - immunology</subject><subject>Leptospira borgpetersenii</subject><subject>Leptospirosis - diagnosis</subject><subject>Leptospirosis - immunology</subject><subject>Leptospirosis - veterinary</subject><subject>Life Sciences</subject><subject>Recombinant Proteins - immunology</subject><subject>Regular Articles</subject><subject>Sensitivity and Specificity</subject><subject>Serologic Tests - veterinary</subject><subject>Veterinary Medicine/Veterinary Science</subject><subject>Zoology</subject><issn>0049-4747</issn><issn>1573-7438</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkkuP1SAYhonROMfRH-BGSdy4QT9oKe3SOfGWnMSFzppQCh3GFirQo_4u_6DUMxpjjK64Pe93e0HoIYVnFEA8T5QyDgQoJ9A2gohbaEe5qIioq_Y22gHUHalFLc7QvZSuAYqqbe6iM9ZAw3nDdujbPsyLiiq7o8HmqKa1bIPHweJodJh755XP-ODGC7zEkI3zWPkBXxmVyUc3TWYo5-xG40mvUjlNKpsvWI3jtOYi_hEtm5TxZ5ev8Ox0DEmHxem_MTZEPDg1-pBc2orow9F5gyez5JAWF7f7--iOVVMyD27Wc3T56uWH_RtyePf67f7FgWhes0y40K2tVJkRs0J3graDtrSyvRYD1aIzjFqoqRHAaMsaawbdM1pX0Pa1Bc6rc_T0FLc0_mkt5cnZJW2mSXkT1iSpYIyKri3T_j9KaSmq5Rv65A_0OqzRl0Y2CjpoQGy56YnaxpWisXKJblbxq6QgN_PlyXxZzJeb-VIUzaObyGs_m-GX4qfbBWAnIJUnP5r4W-p_RH18ElkVpBqjS_LyPQPalO9Ud6Ji1XdLjMbi</recordid><startdate>20151001</startdate><enddate>20151001</enddate><creator>Nagalingam, Mohandoss</creator><creator>Thirumalesh, Sushma Rahim Assadi</creator><creator>Kalleshamurthy, Triveni</creator><creator>Niharika, Nakkala</creator><creator>Balamurugan, Vinayagamurthy</creator><creator>Shome, Rajeswari</creator><creator>Sengupta, Pinaki Prasad</creator><creator>Shome, Bibek Ranjan</creator><creator>Prabhudas, Krishnamsetty</creator><creator>Rahman, Habibur</creator><general>Springer Netherlands</general><general>Springer Nature B.V</general><scope>FBQ</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>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7U7</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope><scope>RC3</scope></search><sort><creationdate>20151001</creationdate><title>Comparative evaluation of recombinant LigB protein and heat-killed antigen-based latex agglutination test with microscopic agglutination test for diagnosis of bovine leptospirosis</title><author>Nagalingam, Mohandoss ; Thirumalesh, Sushma Rahim Assadi ; Kalleshamurthy, Triveni ; Niharika, Nakkala ; Balamurugan, Vinayagamurthy ; Shome, Rajeswari ; Sengupta, Pinaki Prasad ; Shome, Bibek Ranjan ; Prabhudas, Krishnamsetty ; Rahman, Habibur</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c542t-57c8f3a1122f7c9718dcf13fbc7d1c79e21f041e7021826fedcb214308b4f0553</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Animals</topic><topic>Antibodies, Bacterial - immunology</topic><topic>Antigens, Bacterial - immunology</topic><topic>Biomedical and Life Sciences</topic><topic>Cattle</topic><topic>Cattle Diseases - blood</topic><topic>Cattle Diseases - diagnosis</topic><topic>Cattle Diseases - immunology</topic><topic>E coli</topic><topic>Electrophoresis, Polyacrylamide Gel - veterinary</topic><topic>Enzyme-Linked Immunosorbent Assay - veterinary</topic><topic>Escherichia coli</topic><topic>Latex Fixation Tests - veterinary</topic><topic>Leptospira</topic><topic>Leptospira - immunology</topic><topic>Leptospira borgpetersenii</topic><topic>Leptospirosis - diagnosis</topic><topic>Leptospirosis - immunology</topic><topic>Leptospirosis - veterinary</topic><topic>Life Sciences</topic><topic>Recombinant Proteins - immunology</topic><topic>Regular Articles</topic><topic>Sensitivity and Specificity</topic><topic>Serologic Tests - veterinary</topic><topic>Veterinary Medicine/Veterinary Science</topic><topic>Zoology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nagalingam, Mohandoss</creatorcontrib><creatorcontrib>Thirumalesh, Sushma Rahim Assadi</creatorcontrib><creatorcontrib>Kalleshamurthy, Triveni</creatorcontrib><creatorcontrib>Niharika, Nakkala</creatorcontrib><creatorcontrib>Balamurugan, Vinayagamurthy</creatorcontrib><creatorcontrib>Shome, Rajeswari</creatorcontrib><creatorcontrib>Sengupta, Pinaki Prasad</creatorcontrib><creatorcontrib>Shome, Bibek Ranjan</creatorcontrib><creatorcontrib>Prabhudas, Krishnamsetty</creatorcontrib><creatorcontrib>Rahman, Habibur</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>MEDLINE - Academic</collection><collection>Genetics Abstracts</collection><jtitle>Tropical animal health and production</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nagalingam, Mohandoss</au><au>Thirumalesh, Sushma Rahim Assadi</au><au>Kalleshamurthy, Triveni</au><au>Niharika, Nakkala</au><au>Balamurugan, Vinayagamurthy</au><au>Shome, Rajeswari</au><au>Sengupta, Pinaki Prasad</au><au>Shome, Bibek Ranjan</au><au>Prabhudas, Krishnamsetty</au><au>Rahman, Habibur</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparative evaluation of recombinant LigB protein and heat-killed antigen-based latex agglutination test with microscopic agglutination test for diagnosis of bovine leptospirosis</atitle><jtitle>Tropical animal health and production</jtitle><stitle>Trop Anim Health Prod</stitle><addtitle>Trop Anim Health Prod</addtitle><date>2015-10-01</date><risdate>2015</risdate><volume>47</volume><issue>7</issue><spage>1329</spage><epage>1335</epage><pages>1329-1335</pages><issn>0049-4747</issn><eissn>1573-7438</eissn><abstract>This study aimed to develop latex agglutination test (LAT) using recombinant leptospiral immunoglobulin-like protein (LigB) (rLigB) antigen and compare its diagnostic efficacy with LAT using conventional heat-killed leptospiral antigen and microscopic agglutination test (MAT) in diagnosing bovine leptospirosis. The PCR-amplified 1053-bp ligB gene sequences from Leptospira borgpetersenii Hardjo serovar were cloned in pET 32 (a) vector at EcoRI and NotI sites and expressed in BL21 E. coli cells as fusion protein with thioredoxin (−57 kDa) and characterized by SDS-PAGE and immunoblot. Out of 390 serum samples [cattle (n = 214), buffaloes (n = 176)] subjected to MAT, 115 samples showed reciprocal titre≥100 up to 1600 against one or more serovars. For recombinant LigB protein/antigen-based LAT, agglutination was observed in the positive sample, while no agglutination was observed in the negative sample. Similarly, heat-killed leptospiral antigen was prepared from and used in LAT for comparison with MAT. A two-sided contingency table was used for analysis of LAT using both the antigens separately against MAT for 390 serum samples. The sensitivity, specificity and positive and negative predictive values of recombinant LigB LAT were found to be 75.65, 91.27, 78.38 and 89.96 %, respectively, and that of heat-killed antigen-based LAT were 72.17, 89.82, 74.77 and 88.53 %, respectively, in comparison with MAT. This developed test will be an alternative/complementary to the existing battery of diagnostic assays/tests for specific detection of pathogenic Leptospira infection in bovine population.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>26065562</pmid><doi>10.1007/s11250-015-0867-7</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Tropical animal health and production, 2015-10, Vol.47 (7), p.1329-1335 |
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| subjects | Animals Antibodies, Bacterial - immunology Antigens, Bacterial - immunology Biomedical and Life Sciences Cattle Cattle Diseases - blood Cattle Diseases - diagnosis Cattle Diseases - immunology E coli Electrophoresis, Polyacrylamide Gel - veterinary Enzyme-Linked Immunosorbent Assay - veterinary Escherichia coli Latex Fixation Tests - veterinary Leptospira Leptospira - immunology Leptospira borgpetersenii Leptospirosis - diagnosis Leptospirosis - immunology Leptospirosis - veterinary Life Sciences Recombinant Proteins - immunology Regular Articles Sensitivity and Specificity Serologic Tests - veterinary Veterinary Medicine/Veterinary Science Zoology |
| title | Comparative evaluation of recombinant LigB protein and heat-killed antigen-based latex agglutination test with microscopic agglutination test for diagnosis of bovine leptospirosis |
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