{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T01:09:37Z","timestamp":1768266577720,"version":"3.49.0"},"reference-count":67,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,8,26]],"date-time":"2021-08-26T00:00:00Z","timestamp":1629936000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001503","name":"Indian Council of Agricultural Research","doi-asserted-by":"publisher","award":["ICAR Grant number 3021"],"award-info":[{"award-number":["ICAR Grant number 3021"]}],"id":[{"id":"10.13039\/501100001503","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Leptospirosis is responsible for hampering the productivity of swine husbandry worldwide. The aim of this study was to assess the efficacy of bioinformatics tools in predicting the three-dimensional structure and immunogenicity of recombinant LigBCon1-5 (rLigBCon1-5) antigen. A battery of bioinformatics tools such as I-TASSER, ProSA and SAVES v6.0 were used for the prediction and assessment of the predicted structure of rLigBCon1-5 antigen. Bepipred-2.0, DiscoTope v2.0 and ElliPro servers were used to predict linear and conformational epitopes while T-cell epitopes were predicted using NetMHCpan 4.1 and IEDB recommended 2.22 method for MHC Class I and II peptides respectively. The results obtained using various in silico methods were then compared with wet lab experiments comprising of both primary (IgG Dot ELISA Dipstick test) and secondary-binding assays (Latex Agglutination Test [LAT]) to screen 1153 porcine serum samples. The three-dimensional structure of rLigA\/BCon1-5 protein as predicted by I-TASSER was found to be reliable by Ramachandran Plot and ProSA. The ElliPro server suggested 10 and three potential linear and conformational B-cell-epitopes, respectively, on the peptide backbone of the rLigA\/BCon1-5 protein. The DiscoTope prediction server suggested 47 amino acid residues to be part of B-cell antigen. Ten of the most efficient peptides for MHC-I and II grooves were predicted by NetMHCpan 4.1 and IEDB recommended 2.22 method, respectively. Of these, three peptides can serve dual functions as it can fit both MHC I and II grooves, thereby eliciting both humoral-and cell-mediated immune responses. The prediction of these computational approaches proved to be reliable since rLigBCon1-5 antigen-based IgG Dot ELISA Dipstick test and LAT gave results in concordance to gold standard test, the Microscopic Agglutination Test (MAT), for serodiagnosis of leptospirosis. Both the IgG Dot ELISA Dipstick test and LAT were serodiagnostic assays ideally suited for peripheral level of animal health care system as \u201cpoint of care\u201d tests for the detection of porcine leptospirosis.<\/jats:p>","DOI":"10.3390\/pathogens10091082","type":"journal-article","created":{"date-parts":[[2021,8,26]],"date-time":"2021-08-26T09:27:57Z","timestamp":1629970077000},"page":"1082","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Immunoinformatic Study of Recombinant LigA\/BCon1-5 Antigen and Evaluation of Its Diagnostic Potential in Primary and Secondary Binding Tests for Serodiagnosis of Porcine Leptospirosis"],"prefix":"10.3390","volume":"10","author":[{"given":"Sujit Kumar","family":"Behera","sequence":"first","affiliation":[{"name":"Department of Epidemiology & Public Health, Central University of Tamil Nadu, Tiruvarur 610001, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thankappan","family":"Sabarinath","sequence":"additional","affiliation":[{"name":"Clinical Bacteriological Laboratory, Indian Council of Agricultural Research\u2014Indian Veterinary Research Institute, Mukteshwar, Nainital 263138, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8595-8784","authenticated-orcid":false,"given":"Prasanta Kumar K.","family":"Mishra","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary and Animal Sciences, Rajiv Gandhi South Campus, Banaras Hindu University, Mirzapur 231001, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yosef","family":"Deneke","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine, Jimma University, Jimma 378, Ethiopia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ashok","family":"Kumar","sequence":"additional","affiliation":[{"name":"Krishi Bhawan, Indian Council of Agricultural Research, New Delhi 110001, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shanmugam","family":"ChandraSekar","sequence":"additional","affiliation":[{"name":"Biochemistry Laboratory, Indian Council of Agricultural Research\u2014Indian Veterinary Research Institute, Mukteshwar, Nainital 263138, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2346-6028","authenticated-orcid":false,"given":"Kuppusamy","family":"Senthilkumar","sequence":"additional","affiliation":[{"name":"Zoonoses Research Laboratory, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600051, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"MedRam","family":"Verma","sequence":"additional","affiliation":[{"name":"Livestock Economics & Statistics Division, Indian Council of Agricultural Research\u2014Indian Veterinary Research Institute, Bareilly 243122, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1978-2650","authenticated-orcid":false,"given":"Balasubramanian","family":"Ganesh","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Epidemiology, Ayapakkam, Chennai 600077, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amol","family":"Gurav","sequence":"additional","affiliation":[{"name":"Temperate Animal Husbandry Division, ICAR\u2014Indian Veterinary Research Institute (IVRI), Mukteshwar, Nainital 263138, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2183-658X","authenticated-orcid":false,"given":"Abhishek","family":"Hota","sequence":"additional","affiliation":[{"name":"Department of Animal Science, Centurion University of Technology and Management, Paralakhemundi 761211, India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"521","DOI":"10.2478\/bvip-2014-0080","article-title":"Infections of swine caused by Leptospira serovars of serogroup Sejroe possibilities of recognition with the use of PCR","volume":"58","year":"2014","journal-title":"Bull. Vet. Inst. Pulawy"},{"key":"ref_2","first-page":"14","article-title":"Epidemiological Investigation of the Prevalence of Leptospira Spp. in Pigs in Israel","volume":"75","author":"Pozzi","year":"2020","journal-title":"Isr. J. Vet. Med."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Lee, H.S., Thanh, T.L., Ly, N.K., Nguyen-Viet, H., Thakur, K.K., and Grace, D. (2019). Seroprevalence of leptospirosis and Japanese encephalitis in swine in ten provinces of Vietnam. PLoS ONE, 14.","DOI":"10.1101\/584151"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Bertasio, C., Papetti, A., Scaltriti, E., Tagliabue, S., D\u2019Incau, M., and Boniotti, M.B. (2020). Serological survey and molecular typing reveal new Leptospira serogroup Pomona strains among pigs of northern Italy. Pathogens, 9.","DOI":"10.3390\/pathogens9050332"},{"key":"ref_5","unstructured":"Ellis, W.A. (1999). Leptospirosis. Diseases of Swine, Iowa State University Press. [8th ed.]."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1186\/s40813-018-0086-5","article-title":"Passive surveillance of Leptospira infection in swine in Germany","volume":"4","author":"Tschentscher","year":"2018","journal-title":"Porc. Health Manag."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"913","DOI":"10.20506\/rst.35.3.2579","article-title":"Leptospirosis in domestic animals in France: Serological results from 1988 to 2007","volume":"35","year":"2016","journal-title":"Rev. Sci. Tech."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Chadsuthi, S., Bicout, D.J., Wiratsudakul, A., Suwancharoen, D., Petkanchanapong, W., Modchang, C., Triampo, W., Ratanakorn, P., and Chalvet-Monfray, K. (2017). Investigation on predominant Leptospira serovars and its distribution in humans and livestock in Thailand, 2010\u20132015. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0005228"},{"key":"ref_9","first-page":"199","article-title":"Leptospirosis diagnosis: Competancy of various laboratory tests","volume":"8","author":"Budihal","year":"2014","journal-title":"J. Clin. Diagn. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/S0255-0857(21)01991-5","article-title":"Rapid serodiagnosis of leptospirosis by latex agglutination test and flow-through assay","volume":"26","author":"Senthilkumar","year":"2008","journal-title":"Indian J. Med. Microbiol."},{"key":"ref_11","first-page":"391","article-title":"A test strip IgM dot-ELISA assay using leptospiral antigen of endemic strains for serodiagnosis of acute leptospirosis","volume":"88","author":"Tansuphasiri","year":"2005","journal-title":"J. Med. Assoc. Thai."},{"key":"ref_12","first-page":"338","article-title":"IgM Dot-ELISA Assay using prevalent Leptospira strain for diagnosis of leptospirosis","volume":"2","author":"Shekatkar","year":"2010","journal-title":"Int. J. Collab. Res. Intern. Med. Public Health"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.proche.2016.01.004","article-title":"Recombinant LipL32 protein for leptospirosis detection in Indonesia","volume":"18","author":"Sumarningsih","year":"2016","journal-title":"Procedia Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"99","DOI":"10.4142\/jvs.2012.13.1.99","article-title":"Evaluation and comparison of native and recombinant LipL21 protein based ELISAs for diagnosis of bovine leptospirosis","volume":"13","author":"Joseph","year":"2012","journal-title":"J. Vet. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1179\/136485908X355229","article-title":"Serodiagnosis of severe leptospirosis: Evaluation of ELISA based on the recombinant OmpL1 or LipL41 antigens of Leptospira interrogans serovar autumnalis","volume":"102","author":"Natarajaseenivasan","year":"2008","journal-title":"Ann. Trop. Med. Parasitol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Ye, C., Yan, W., Xiang, H., He, H., Yang, M., Ijaz, M., Useh, N., Hsieh, C.L., McDonough, P.L., and McDonough, S.P. (2014). Recombinant Antigens rLipL21, rLoa22, rLipL32 and rLigACon4-8 for Serological Diagnosis of Leptospirosis by Enzyme-Linked Immunosorbent Assays in Dogs. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0111367"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1616","DOI":"10.1128\/CVI.00345-07","article-title":"Use of recombinant antigens to detect antibodies against Mycoplasma suis with correlation of serological results to hematological findings","volume":"14","author":"Hoelzle","year":"2007","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1711","DOI":"10.1128\/CVI.00189-08","article-title":"Recombinant Multiepitope Protein for Diagnosis of Leptospirosis","volume":"15","author":"Lin","year":"2008","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1046\/j.1365-2958.2003.03619.x","article-title":"Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily","volume":"49","author":"Matsunaga","year":"2003","journal-title":"Mol. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1768","DOI":"10.1038\/s41598-021-81400-7","article-title":"Genetic manipulation of pathogenic Leptospira: CRISPR interference (CRISPRi)-mediated gene silencing and rapid mutant recovery at 37 \u00b0C","volume":"11","author":"Fernandes","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.imlet.2016.03.005","article-title":"Acquisition of negative complement regulators by the saprophyte Leptospira bifexa expressing LigA or LigB confers enhanced survival in human serum","volume":"173","author":"Fraga","year":"2016","journal-title":"Immunol. Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1528","DOI":"10.1128\/JCM.02344-06","article-title":"Leptospira immunoglobulin-like proteins as a serodiagnostic marker for acute leptospirosis","volume":"45","author":"Croda","year":"2007","journal-title":"J. Clin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1099\/jmm.0.009175-0","article-title":"Distribution of the leptospiral immunoglobulin-like (lig) genes in pathogenic Leptospira species and application of ligB to typing leptospiral isolates","volume":"58","author":"Cerqueira","year":"2009","journal-title":"J. Med. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1099\/jmm.0.45568-0","article-title":"Expression of leptospiral immunoglobulin-like protein by Leptospira interrogans and evaluation of its diagnostic potential in a kinetic ELISA","volume":"53","author":"Palaniappan","year":"2004","journal-title":"J. Med. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1038\/nprot.2010.5","article-title":"I-TASSER: A unified platform for automated protein structure and function prediction","volume":"5","author":"Roy","year":"2010","journal-title":"Nat. Protoc."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1002\/prot.10141","article-title":"Local energy landscape flattening: Parallel hyperbolic Monte Carlo sampling of protein folding","volume":"48","author":"Zhang","year":"2002","journal-title":"Proteins"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1002\/prot.21702","article-title":"Template-based modeling and free modeling by I-TASSER in CASP7","volume":"69","author":"Zhang","year":"2007","journal-title":"Proteins Struct. Funct. Bioinf."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1002\/j.1460-2075.1986.tb04226.x","article-title":"Location of \u2018continuous\u2019 antigenic determinants in the protruding regions of proteins","volume":"5","author":"Thornton","year":"1986","journal-title":"EMBO J."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Eswar, N.W.B., Marti-Renom, M.A., Madhusudhan, M.S., Eramian, D., Shen, M.Y., Pieper, U., and Sali, A. (2006). Comparative Protein Structure Modeling with MODELLER.InCurrent Protocols in Bioinformatics, Wiley.","DOI":"10.1002\/0471250953.bi0506s15"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"5.6.1","DOI":"10.1002\/cpbi.3","article-title":"Comparative Protein Structure Modeling using MODELLER","volume":"54","author":"Webb","year":"2016","journal-title":"Curr. Protoc. Bioinform."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Ponomarenko, J., Bui, H.H., Li, W., Fusseder, N., Bourne, P.E., Sette, A., and Peters, B. (2008). ElliPro: A new structure-based tool for the prediction of antibody epitopes. BMC Bioinform., 9.","DOI":"10.1186\/1471-2105-9-514"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2558","DOI":"10.1110\/ps.062405906","article-title":"Prediction of residues in discontinuous B-cell epitopes using protein 3D structures","volume":"15","author":"Andersen","year":"2006","journal-title":"Protein Sci."},{"key":"ref_33","unstructured":"Grande, T.L. (2021, June 06). Calculating the Sample Size with a Finite Population in Excel (Video File). Available online: https:\/\/www.youtube.com\/watch?v=gLD4tENS82c&t=189s."},{"key":"ref_34","unstructured":"USDA (United States Department of Agriculture) (1987). National Veterinary Services Laboratories. Microtitre technique for detection of Leptospira antibodies. Proc. Annu. Meet. US Anim. Health Assoc., 91, 65\u201373."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1007\/s13213-019-01516-0","article-title":"In silico characterization of Leptospira interrogans DNA ligase A and delineation of its antimicrobial stretches","volume":"69","author":"Mishra","year":"2019","journal-title":"Ann. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"W407","DOI":"10.1093\/nar\/gkm290","article-title":"ProSA-web: Interactive web service for the recognition of errors in three-dimensional structures of proteins","volume":"35","author":"Wiederstein","year":"2007","journal-title":"Nucleic Acids Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1126\/science.1853201","article-title":"A method to identify protein sequences that fold into a known three-dimensional structure","volume":"253","author":"Bowie","year":"1991","journal-title":"Science"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1038\/356083a0","article-title":"Assessment of protein models with three-dimensional profiles","volume":"356","author":"Bowie","year":"1992","journal-title":"Nature"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1511","DOI":"10.1002\/pro.5560020916","article-title":"Verification of protein structures: Patterns of non bonded atomic interactions","volume":"2","author":"Colovos","year":"1993","journal-title":"Protein Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1006\/jmbi.1996.0628","article-title":"Deviations from Standard Atomic Volumes as a Quality Measure for Protein Crystal Structures","volume":"264","author":"Pontius","year":"1996","journal-title":"J. Mol. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Kringelum, J.V., Lundegaard, C., Lund, O., and Nielsen, M. (2012). Reliable B Cell Epitope Predictions: Impacts of Method Development and Improved Benchmarking. PLoS Comput. Biol., 8.","DOI":"10.1371\/journal.pcbi.1002829"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"W449","DOI":"10.1093\/nar\/gkaa379","article-title":"NetMHCpan-4.1 and NetMHCIIpan-4.0: Improved predictions of MHC antigen presentation by concurrent motif deconvolution and integration of MS MHC eluted ligand data","volume":"48","author":"Reynisson","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.mcp.2014.01.001","article-title":"Evaluation of recombinant LigB antigen-based indirect ELISA and latex agglutination test for the serodiagnosis of bovine leptospirosis in India","volume":"28","author":"Deneke","year":"2014","journal-title":"Mol. Cell. Probes"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of structural proteins during the assembly of the head of bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4350","DOI":"10.1073\/pnas.76.9.4350","article-title":"Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications","volume":"76","author":"Towbin","year":"1979","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1007\/s11259-006-3364-7","article-title":"Recombinant antigen-based Latex Agglutination Test for Rapid Serodiagnosis of leptospirosis","volume":"31","author":"Dey","year":"2007","journal-title":"Vet. Res. Commun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1272","DOI":"10.1128\/JCM.38.3.1272-1275.2000","article-title":"Simple latex agglutination assay for rapid serodiagnosis of human leptospirosis","volume":"38","author":"Smits","year":"2000","journal-title":"J. Clin. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"469","DOI":"10.20506\/rst.17.2.1119","article-title":"Validation of serological assays for diagnosis of infectious diseases","volume":"17","author":"Jacobson","year":"1998","journal-title":"Rev. Sci. Tech. Off. Int. Epiz."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Ngugi, J.N., F\u00e8vre, E.M., Mgode, G.F., Obonyo, M., Mhamphi, G.G., Otieno, C.A., and Cook, E.A.J. (2019). Seroprevalence and associated risk factors of leptospirosis in slaughter pigs; a neglected public health risk, western Kenya. BMC Vet. Res., 15.","DOI":"10.1186\/s12917-019-2159-3"},{"key":"ref_50","first-page":"693","article-title":"Deciphering leptospirosis-a diagnostic mystery, an insight","volume":"4","author":"Bhatia","year":"2015","journal-title":"J. Med Res. Health Sci."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.4269\/ajtmh.14-0551","article-title":"Prevalence of Endemic Pig-Associated Zoonoses in Southeast Asia: A Review of Findings from the Lao People\u2019s Democratic Republic","volume":"92","author":"Okello","year":"2015","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_52","unstructured":"Faine, S.B., Adler, B., Bolin, C., and Perolat, P. (1999). Leptospira and Leptospirosis, MediSci. [2nd ed.]."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"17","DOI":"10.14202\/vetworld.2014.17-20","article-title":"Evaluation of recombinant LipL32 based latex agglutination test for serodiagnosis of porcine leptospirosis","volume":"7","author":"Behera","year":"2014","journal-title":"Vet. World"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1590\/S0103-84782002000400011","article-title":"Most frequent serovars of leptospires in serological tests of buffaloes, sheeps, goats, horses, swines and dogs from several Brazilian states","volume":"32","author":"Favero","year":"2002","journal-title":"Cienc. Rural."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1111\/j.1865-1682.2012.01320.x","article-title":"Prevalence and risk factors associated with leptospira spp. infection in technified swine farms in the state of Alagoas, Brazil risk factors associated with leptospira spp. in swine farms","volume":"60","author":"Mota","year":"2013","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1590\/S1517-83822002000200016","article-title":"Leptospirosis in slaughtered sows: Serological and histopathological investigation","volume":"33","author":"Delbem","year":"2002","journal-title":"Braz. J. Microbiol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"5249","DOI":"10.1021\/bi500669u","article-title":"NMR Solution Structure of the Terminal Immunoglobulin-like Domain from the Leptospira Host-Interacting Outer Membrane Protein, LigB","volume":"53","author":"Ptak","year":"2014","journal-title":"Biochemistry"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1002\/prot.24723","article-title":"Solution structure of a bacterial immunoglobulin-like domain of the outer membrane protein (LigB) from Leptospira","volume":"83","author":"Mei","year":"2015","journal-title":"Proteins"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"e30051","DOI":"10.7554\/eLife.30051","article-title":"Extended low-resolution structure of a Leptospira antigen offers high bactericidal antibody accessibility amenable to vaccine design","volume":"6","author":"Hsieh","year":"2017","journal-title":"eLife"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Wiltgen, M. (2019). Algorithms for Structure Comparison and Analysis: Homology Modelling of Proteins InEncyclopedia of Bioinformatics and Computational Biology, Elsevier.","DOI":"10.1016\/B978-0-12-809633-8.20484-6"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"672","DOI":"10.2174\/092986607781483921","article-title":"Structural compromise of disallowed conformations in peptide and protein structures","volume":"14","author":"Ramakrishnan","year":"2007","journal-title":"Protein Pept. Lett."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1002\/prot.340170404","article-title":"Recognition of errors in three-dimensional structures of proteins","volume":"17","author":"Sippl","year":"1993","journal-title":"Proteins"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1007\/s00251-013-0720-y","article-title":"NetMHCIIpan-3.0, a common pan-specific MHC class II prediction method including all three human MHC class II isotypes, HLA-DR, HLA-DP and HLA-DQ","volume":"65","author":"Karosiene","year":"2013","journal-title":"Immunogenetics"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.tim.2011.11.002","article-title":"The pig: A model for human infectious diseases","volume":"20","author":"Meurens","year":"2012","journal-title":"Trends Microbiol."},{"key":"ref_65","first-page":"149","article-title":"Genomic Analysis of a New Serovar of Leptospira weilii Serogroup Manhao","volume":"8","author":"Xu","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.jmii.2013.03.001","article-title":"Laboratory diagnosis of leptospirosis: A challenge","volume":"46","author":"Musso","year":"2013","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"1853","DOI":"10.1128\/JCM.44.5.1853-1855.2006","article-title":"Evaluation of a commercial latex agglutination assay for serological diagnosis of leptospirosis","volume":"44","author":"Glass","year":"2006","journal-title":"J. Clin. Microbiol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/9\/1082\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:52:10Z","timestamp":1760165530000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/9\/1082"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,26]]},"references-count":67,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["pathogens10091082"],"URL":"https:\/\/doi.org\/10.3390\/pathogens10091082","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,8,26]]}}}