{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T05:17:07Z","timestamp":1775193427173,"version":"3.50.1"},"reference-count":61,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"2","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,8,2]]},"abstract":"<jats:p>Leptospirosis is a neglected zoonotic disease with worldwide endemicity and continues to be a significant public health burden on resource-limited populations. Previously, we produced three highly purified recombinant antigens (rLipL32, rLipL41, and rLigA-Rep) and evaluated their performance of detecting <jats:italic>Leptospira<\/jats:italic>-specific antibodies in enzyme-linked immunosorbent assay (ELISA) as compared with the microscopic agglutination test (MAT). The overall sensitivity of this assay approached 90%. Recently, another recombinant antigen (rLigB-Rep) was prepared. We tested each individual antigen and a 1:1:1:1 mixture of these four antigens for the detection of <jats:italic>Leptospira<\/jats:italic>-specific antibodies in ELISA. The performance of these recombinant antigens was evaluated with a much larger febrile patient panel (337 MAT-confirmed positive sera and 92 MAT-negative sera from febrile patients). Combining the detection results of immunoglobulin M and immunoglobulin G from these four individual antigens, the overall sensitivity was close to 90% but the specificity was only 66%, based on the MAT reference method. The overall sensitivity and specificity of the four-antigen mixture were 82% and 86%, respectively. The mixture of four antigens also exhibited a broader reactivity with MAT-positive samples of 18 serovars from six major pathogenic <jats:italic>Leptospira<\/jats:italic> species. Given the limitations of MAT, the data were further analyzed by Bayesian latent class model, showing that ELISA using a 1:1:1:1 mixture still maintained high sensitivity (79%) and specificity (88%) as compared with the sensitivity (90%) and specificity (83%) of MAT. Therefore, ELISA using a mixture of these four antigens could be a very useful test for seroprevalence studies.<\/jats:p>","DOI":"10.4269\/ajtmh.17-0057","type":"journal-article","created":{"date-parts":[[2018,6,26]],"date-time":"2018-06-26T07:01:01Z","timestamp":1529996461000},"page":"266-274","source":"Crossref","is-referenced-by-count":12,"title":["An Improved Enzyme-Linked Immunoassay for the Detection of Leptospira-Specific Antibodies"],"prefix":"10.4269","volume":"99","author":[{"given":"Hua-Wei","family":"Chen","sequence":"first","affiliation":[{"name":"Naval Medical Research Center, Silver Spring, Maryland;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heather","family":"Lukas","sequence":"additional","affiliation":[{"name":"Naval Medical Research Center, Silver Spring, Maryland;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kira","family":"Becker","sequence":"additional","affiliation":[{"name":"Naval Medical Research Center, Silver Spring, Maryland;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giulia","family":"Weissenberger","sequence":"additional","affiliation":[{"name":"Naval Medical Research Center, Silver Spring, Maryland;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric S.","family":"Halsey","sequence":"additional","affiliation":[{"name":"Naval Medical Research Unit No. 6, Lima, Peru;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolina","family":"Guevara","sequence":"additional","affiliation":[{"name":"Naval Medical Research Unit No. 6, Lima, Peru;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Enrique","family":"Canal","sequence":"additional","affiliation":[{"name":"Naval Medical Research Unit No. 6, Lima, Peru;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric","family":"Hall","sequence":"additional","affiliation":[{"name":"Naval Medical Research Center, Silver Spring, Maryland;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryan C.","family":"Maves","sequence":"additional","affiliation":[{"name":"Naval Medical Research Unit No. 6, Lima, Peru;"},{"name":"Naval Medical Center San Diego, San Diego, California;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Drake H.","family":"Tilley","sequence":"additional","affiliation":[{"name":"Naval Medical Center San Diego, San Diego, California;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lynn","family":"Kuo","sequence":"additional","affiliation":[{"name":"Department of Statistics, University of Connecticut, Storrs, Connecticut"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tadeusz J.","family":"Kochel","sequence":"additional","affiliation":[{"name":"Naval Medical Research Center, Silver Spring, Maryland;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei-Mei","family":"Ching","sequence":"additional","affiliation":[{"name":"Naval Medical Research Center, Silver Spring, Maryland;"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2795","reference":[{"key":"b1","series-title":"Int J Infect Dis","first-page":"351","article-title":"The globalization of leptospirosis: worldwide incidence trends","volume":"12","author":"Pappas","year":"2008"},{"key":"b2","series-title":"Clin Microbiol Rev","first-page":"296","article-title":"Leptospirosis","volume":"14","author":"Levett","year":"2001"},{"key":"b3","series-title":"PLoS Negl Trop Dis","first-page":"e0003898","article-title":"Global morbidity and mortality of leptospirosis: a systematic review","volume":"9","author":"Costa","year":"2015"},{"key":"b4","series-title":"Ethn Dis","first-page":"37","article-title":"Leptospirosis: a worldwide resurgent zoonosis and important cause of acute renal failure and death in developing nations","volume":"19","author":"Cruz","year":"2009"},{"key":"b5","series-title":"PLoS Negl Trop Dis","first-page":"579","article-title":"Increasing trends of leptospirosis in northern India: a clinico-epidemiological study","volume":"4","author":"Sethi","year":"2010"},{"key":"b6","series-title":"BMC Infect Dis","first-page":"147","article-title":"Leptospirosis in the Asia Pacific region","volume":"9","author":"Victoriano","year":"2009"},{"key":"b7","series-title":"Malays J Pathol","first-page":"1","article-title":"Leptospirosis: a re-emerging infection","volume":"33","author":"Lim","year":"2011"},{"key":"b8","series-title":"Emerg Infect Dis","first-page":"1678","article-title":"Leptospirosis as frequent cause of acute febrile illness in southern Sri Lanka","volume":"17","author":"Reller","year":"2011"},{"key":"b9","series-title":"J Clin Microbiol","first-page":"307","article-title":"Human Leptospira isolates circulating in Mayotte (Indian Ocean) have unique serological and molecular features","volume":"50","author":"Bourhy","year":"2012"},{"key":"b10","series-title":"Rev Soc Bras Med Trop","first-page":"100","article-title":"Geographic analysis on the occurrence of human and canine leptospirosis in the city of Maring\u00e1, state of Paran\u00e1, Brazil","volume":"45","author":"Fonzar","year":"2012"},{"key":"b11","series-title":"Am J Trop Med Hyg","first-page":"309","article-title":"Leptospirosis in American Samoa 2010: epidemiology, environmental drivers, and the management of emergence","volume":"86","author":"Lau","year":"2012"},{"key":"b12","series-title":"Microbes Infect","first-page":"317","article-title":"Leptospirosis prevalence in Chinese populations in the last two decades","volume":"14","author":"Zhang","year":"2012"},{"key":"b13","series-title":"Curr Opin Infect Dis","first-page":"376","article-title":"Leptospirosis","volume":"18","author":"McBride","year":"2005"},{"key":"b14","series-title":"Nat Rev Microbiol","first-page":"736","article-title":"Leptospira: the dawn of the molecular genetics era for an emerging zoonotic pathogen","volume":"7","author":"Ko","year":"2009"},{"key":"b15","series-title":"Lancet","first-page":"535","article-title":"Leptospirosis associated with outbreak of acute febrile illness and pulmonary haemorrhage, Nicaragua, 1995. The epidemic working group at ministry of health in Nicaragua","volume":"347","author":"Zaki","year":"1996"},{"key":"b16","series-title":"Emerg Infect Dis","first-page":"505","article-title":"Leptospirosis-associated severe pulmonary hemorrhagic syndrome, Salvador, Brazil","volume":"14","author":"Gouveia","year":"2008"},{"key":"b17","series-title":"Emerg Infect Dis","first-page":"834","article-title":"Pulmonary involvement and leptospirosis, Greece","volume":"15","author":"Papa","year":"2009"},{"key":"b18","series-title":"PLoS Negl Trop Dis","first-page":"e2324","article-title":"Etiology of severe non-malaria febrile illness in northern Tanzania: a prospective cohort study","volume":"7","author":"Crump","year":"2013"},{"key":"b19","series-title":"Emerg Infect Dis","first-page":"766","article-title":"Leptospirosis during dengue outbreak, Bangladesh","volume":"11","author":"LaRocque","year":"2005"},{"key":"b20","series-title":"Int J Antimicrob Agents","first-page":"S5","article-title":"Estimating the burden of human leptospirosis","volume":"36","author":"Abela-Ridder","year":"2010"},{"key":"b21","series-title":"PLoS Negl Trop Dis","first-page":"e2589","article-title":"Estimating leptospirosis incidence using hospital-based surveillance and a population-based health care utilization survey in Tanzania","volume":"7","author":"Biggs","year":"2013"},{"key":"b22","first-page":"963","author":"Levett","year":"2007"},{"key":"b23","series-title":"Lancet Infect Dis","first-page":"757","article-title":"Leptospirosis: a zoonotic disease of global importance","volume":"3","author":"Bharti","year":"2003"},{"key":"b24","series-title":"J Clin Microbiol","first-page":"3303","article-title":"Evaluation of recombinant Leptospira antigen-based enzyme-linked immunosorbent assays for the serodiagnosis of leptospirosis","volume":"39","author":"Flannery","year":"2001"},{"key":"b25","series-title":"J Microbiol Methods","first-page":"73","article-title":"Recombinant ligA for leptospirosis diagnosis and ligA among the Leptospira spp. clinical isolates","volume":"72","author":"Srimanote","year":"2008"},{"key":"b26","series-title":"Trans R Soc Trop Med Hyg","first-page":"289","article-title":"Leptospirosis serodiagnosis by ELISA based on recombinant outer membrane protein","volume":"105","author":"Chalayon","year":"2011"},{"key":"b27","series-title":"Am J Trop Med Hyg","first-page":"1088","article-title":"Detection of Leptospira-specific antibodies using a recombinant antigen-based enzyme-linked immunosorbent assay","volume":"89","author":"Chen","year":"2013"},{"key":"b28","series-title":"PLoS Negl Trop Dis","first-page":"e0003879","article-title":"Diagnostic accuracy of recombinant immunoglobulin-like protein a-based IgM ELISA for the early diagnosis of leptospirosis in the Philippines","volume":"9","author":"Kitashoji","year":"2015"},{"key":"b29","series-title":"J Mol Biol","first-page":"113","article-title":"Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes","volume":"189","author":"Studier","year":"1986"},{"key":"b30","series-title":"Mol Microbiol","first-page":"929","article-title":"Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily","volume":"49","author":"Matsunaga","year":"2003"},{"key":"b31","series-title":"J Immunol Methods","first-page":"35","article-title":"A statistically defined endpoint titer determination method for immunoassays","volume":"221","author":"Frey","year":"1998"},{"key":"b32","series-title":"Am J Epidemiol","first-page":"263","article-title":"Bayesian estimation of disease prevalence and the parameters of diagnostic tests in the absence of a gold standard","volume":"141","author":"Joseph","year":"1995"},{"key":"b33","series-title":"Biometrics","first-page":"158","article-title":"Bayesian approaches to modeling the conditional dependence between multiple diagnostic tests","volume":"57","author":"Dendukuri","year":"2001"},{"key":"b34","series-title":"Biometrics","first-page":"959","article-title":"The effect of conditional dependence on the evaluation of diagnostic tests","volume":"41","author":"Vacek","year":"1985"},{"key":"b35","series-title":"Prev Vet Med","first-page":"145","article-title":"Estimation of diagnostic-test sensitivity and specificity through Bayesian modeling","volume":"68","author":"Branscum","year":"2005"},{"key":"b36","series-title":"Infect Immun","first-page":"5924","article-title":"Cloning and molecular characterization of an immunogenic LigA protein of Leptospira interrogans","volume":"70","author":"Palaniappan","year":"2002"},{"key":"b37","series-title":"Vaccine","first-page":"1545","article-title":"Leptospiral immunoglobulin-like proteins elicit protective immunity","volume":"22","author":"Koizumi","year":"2004"},{"key":"b38","series-title":"Infect Genet Evol","first-page":"196","article-title":"Genetic diversity of the leptospiral immunoglobulin-like (Lig) genes in pathogenic Leptospira spp","volume":"9","author":"Mcbride","year":"2009"},{"key":"b39","author":"Agresti","year":"1996"},{"key":"b40","series-title":"Infect Immun","first-page":"4958","article-title":"Leptospiral proteins recognized during the humoral immune response to leptospirosis in humans","volume":"69","author":"Guerreiro","year":"2001"},{"key":"b41","series-title":"J Med Microbiol","first-page":"1259","article-title":"Percutaneous exposure resulting in laboratory-acquired leptospirosis\u2014a case report","volume":"53","author":"Sugunan","year":"2004"},{"key":"b42","series-title":"Clin Diagn Lab Immunol","first-page":"166","article-title":"Lateral-flow assay for rapid serodiagnosis of human leptospirosis","volume":"8","author":"Smits","year":"2001"},{"key":"b43","series-title":"J Clin Microbiol","first-page":"1464","article-title":"Evaluation of eight rapid screening tests for acute leptospirosis in Hawaii","volume":"40","author":"Effler","year":"2002"},{"key":"b44","series-title":"Am J Trop Med Hyg","first-page":"745","article-title":"Evaluation of two enzyme-linked immunosorbent assay methods for detection of immunoglobulin M antibodies in acute leptospirosis","volume":"66","author":"Levett","year":"2002"},{"key":"b45","series-title":"Epidemiol Infect","first-page":"617","article-title":"Evaluation of Lepto Dri Dot as a rapid test for the diagnosis of leptospirosis","volume":"129","author":"Vijayachari","year":"2002"},{"key":"b46","series-title":"J Clin Microbiol","first-page":"803","article-title":"Evaluation of four commercially available rapid serologic tests for diagnosis of leptospirosis","volume":"41","author":"Bajani","year":"2003"},{"key":"b47","series-title":"J Med Microbiol","first-page":"897","article-title":"Field application of lepto lateral flow for rapid diagnosis of leptospirosis","volume":"52","author":"Sehgal","year":"2003"},{"key":"b48","series-title":"Curr Opin Infect Dis","first-page":"376","article-title":"Leptospirosis","volume":"18","author":"McBride","year":"2005"},{"key":"b49","series-title":"Clin Vaccine Immunol","first-page":"1166","article-title":"Limited diagnostic capacities of two commercial assays for the detection of Leptospira immunoglobulin M antibodies in Laos","volume":"3","author":"Blacksell","year":"2006"},{"key":"b50","series-title":"Am J Trop Med Hyg","first-page":"524","article-title":"Accuracy of a commercial IgM ELISA for the diagnosis of human leptospirosis in Thailand","volume":"86","author":"Desakorn","year":"2012"},{"key":"b51","series-title":"PLoS Negl Trop Dis","first-page":"e2499","article-title":"Identification of seroreactive proteins of Leptospira interrogans serovar copenhageni using a high-density protein microarray approach","volume":"7","author":"Lessa-Aquino","year":"2013"},{"key":"b52","series-title":"J Med Microbiol","first-page":"1173","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"},{"key":"b53","series-title":"Infect Immun","first-page":"2276","article-title":"The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection","volume":"68","author":"Haake","year":"2000"},{"key":"b54","series-title":"Gen Tijdschr Ned Ind","first-page":"226","article-title":"Een serologish afwijkende leptospira-stain uit der nier eener vleermuis","volume":"79","author":"Collier","year":"1939"},{"key":"b55","series-title":"PLoS Negl Trop Dis","first-page":"e213","article-title":"Human leptospirosis caused by a new, antigenically unique Leptospira associated with a Rattus species reservoir in the Peruvian Amazon","volume":"2","author":"Matthias","year":"2008"},{"key":"b56","series-title":"Emerg Infect Dis","first-page":"1016","article-title":"Environmental exposure and leptospirosis, Peru","volume":"10","author":"Johnson","year":"2004"},{"key":"b57","series-title":"Rev Peru Med Exp Salud Publica","first-page":"195","article-title":"Leptospirosis seroprevalence and associated features in rice farmers of tropical region of Peru","volume":"31","author":"Alarc\u00f3n-Villaverde","year":"2014"},{"key":"b58","series-title":"Am J Trop Med Hyg","first-page":"731","article-title":"Assessment of the efficacy of an IgM-elisa and microscopic agglutination test (MAT) in the diagnosis of acute leptospirosis","volume":"61","author":"Cumberland","year":"1999"},{"key":"b59","series-title":"Clin Infect Dis","first-page":"322","article-title":"Fool\u2019s gold: why imperfect reference tests are undermining the evaluation of novel diagnostics: a reevaluation of 5 diagnostic tests for leptospirosis","volume":"55","author":"Limmathurotsakul","year":"2012"},{"key":"b60","series-title":"PLoS 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