{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T22:13:46Z","timestamp":1790806426003,"version":"4.1.0"},"reference-count":69,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2025,6,24]],"date-time":"2025-06-24T00:00:00Z","timestamp":1750723200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Although tissue aspiration procedures (TAPs) are considered the gold standard for visceral leishmaniasis (VL) diagnosis, they often fail to detect disease progression before the amastigote-demonstrable phase. To overcome this limitation, a primary direct agglutination test (DAT) was developed using intact <jats:italic>Leishmania donovani<\/jats:italic> promastigotes initially treated with trypsin as the antigen. To enhance the exposure of specific epitopes on the promastigote surface and antibody-binding sites, alternative proteolytic agents were evaluated and incorporated into antigen processing and test execution. This approach led to significant or complete inhibition of agglutination in most known cross-reacting disorders. The LQ-DAT consistently demonstrated highly reproducible results across diverse geographical regions, regardless of the <jats:italic>L. donovani<\/jats:italic> sub-species or strain. To facilitate routine implementation and local production, the LQ-DAT processing know-how was disseminated to all major VL-endemic areas. Sensitivities comparable to TAP outcomes were demonstrated in 2,224 of 2,697 VL cases successfully diagnosed and treated over 35 years of routine, epidemic, and outbreak evaluations. Notably, 473 (17.5%) of these cases were symptomatic, with TAP-negative but LQ-DAT-positive results, and responded favorably (98.0%\u2212100%) to specific treatment. Given the lower sensitivity also demonstrated by LQ-DAT, TAP does not meet the criteria for classification as the gold-standard VL diagnostic. Consequently, a positive response to specific anti-leishmanial treatment is recommended as a benchmark for diagnostic reliability. Beyond its advantage in detecting VL at earlier stages compared to TAP, the improved LQ-DAT described here also exhibited feasibility and stability required for local production in low-resource settings.<\/jats:p>","DOI":"10.1371\/journal.pone.0319118","type":"journal-article","created":{"date-parts":[[2025,6,24]],"date-time":"2025-06-24T17:36:22Z","timestamp":1750786582000},"page":"e0319118","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Direct agglutination test: Evolution and significance as a simple and safe alternative to tissue aspiration procedures in the diagnosis of visceral leishmaniasis"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3099-300X","authenticated-orcid":true,"given":"Abdallah","family":"el Harith","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2017-2331","authenticated-orcid":true,"given":"Hussam Ali","family":"Osman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Durria","family":"Mansour","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saul J.","family":"Semiao-Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0156-7346","authenticated-orcid":true,"given":"Elfadil","family":"Abass","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,6,24]]},"reference":[{"key":"pone.0319118.ref001","doi-asserted-by":"crossref","first-page":"723","DOI":"10.4269\/ajtmh.1962.11.723","article-title":"Leishmaniasis in the Sudan republic. VI. Clinical and laboratory aspects of kala\u2010azar in hospitalized patients from Upper Nile province","volume":"11","author":"PFD van Peenen","year":"1962","journal-title":"Am J Trop Med Hyg"},{"issue":"1","key":"pone.0319118.ref002","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/0035-9203(88)90265-9","article-title":"Visceral leishmaniasis in the Sudan: comparative parasitological methods of diagnosis","volume":"82","author":"M Siddig","year":"1988","journal-title":"Trans R Soc Trop Med Hyg"},{"issue":"2","key":"pone.0319118.ref003","first-page":"125","article-title":"Splenic aspiration; experience in Kenya","volume":"35","author":"PA Kager","year":"1983","journal-title":"Trop Geogr Med"},{"key":"pone.0319118.ref004","doi-asserted-by":"crossref","first-page":"811","DOI":"10.4269\/ajtmh.2005.72.811","article-title":"Sensitivity of bone marrow aspirates in the diagnosis of visceral leishmaniasis","volume":"72","author":"MR da Silva","year":"2005","journal-title":"Am J Trop Med Hyg"},{"issue":"5","key":"pone.0319118.ref005","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1016\/0035-9203(92)90086-R","article-title":"Kala-azar: a comparative study of parasitological methods and the direct agglutination test in diagnosis","volume":"86","author":"EE Zijlstra","year":"1992","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"pone.0319118.ref006","first-page":"391","article-title":"Potential of direct agglutination test (DAT) for detection of visceral leishmaniasis in a known endemic area in Sudan","volume":"4","author":"AA Abdel-Hameed","year":"1989","journal-title":"Trop Med Parasitol"},{"issue":"10","key":"pone.0319118.ref007","doi-asserted-by":"crossref","first-page":"2454","DOI":"10.1128\/jcm.35.10.2454-2457.1997","article-title":"Evaluation of PCR for diagnosis of visceral leishmaniasis","volume":"35","author":"OF Osman","year":"1997","journal-title":"J Clin Microbiol"},{"issue":"4","key":"pone.0319118.ref008","doi-asserted-by":"crossref","first-page":"689","DOI":"10.4269\/ajtmh.2007.76.689","article-title":"Utility of lymph node aspiration in the diagnosis of visceral leishmaniasis in Sudan","volume":"76","author":"ZOE Babiker","year":"2007","journal-title":"Am J Trop Med Hyg"},{"issue":"5","key":"pone.0319118.ref009","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1080\/00034983.1984.11811853","article-title":"A complement fixation test for visceral leishmaniasis using homologous parasite antigen I","volume":"78","author":"WT Hockmeyer","year":"1984","journal-title":"Ann Trop Med Parasitol"},{"issue":"3","key":"pone.0319118.ref010","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1080\/00034983.1978.11719308","article-title":"The micro-ELISA technique in the serodiagnosis of visceral leishmaniasis","volume":"72","author":"M Hommel","year":"1978","journal-title":"Ann Trop Med Parasitol"},{"issue":"3","key":"pone.0319118.ref011","doi-asserted-by":"crossref","first-page":"296","DOI":"10.4269\/ajtmh.1972.21.296","article-title":"Serodiagnosis of American leishmaniasis by indirect fluorescent antibody test","volume":"21","author":"BC Walton","year":"1972","journal-title":"Am J Trop Med Hyg"},{"issue":"2","key":"pone.0319118.ref012","first-page":"247","article-title":"A serological study on human Trypanosoma rhodesiense infections using a micro-scale enzyme linked immunosorbent assay","volume":"26","author":"A Voller","year":"1975","journal-title":"Trop Med Parasitol"},{"issue":"5","key":"pone.0319118.ref013","doi-asserted-by":"crossref","first-page":"943","DOI":"10.4269\/ajtmh.1983.32.943","article-title":"An enzyme-linked immunosorbent assay (ELISA) for field diagnosis of visceral leishmaniasis","volume":"32","author":"M Ho","year":"1983","journal-title":"Am J Trop Med Hyg"},{"key":"pone.0319118.ref014","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1111\/j.1699-0463.1993.tb00158.x","article-title":"Measurement of serum antibodies against native Leishmania gp63 distinguishes between ongoing and previous L. donovani infection","volume":"101","author":"EA Okong\u2019o-Odera","year":"1993","journal-title":"APMIS"},{"key":"pone.0319118.ref015","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/S0001-706X(02)00043-8","article-title":"Antigenuria in visceral leishmaniasis: detection and partial characterisation of a carbohydrate antigen","volume":"82","author":"B Sarkari","year":"2002","journal-title":"Acta Tropica"},{"issue":"2","key":"pone.0319118.ref016","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1073\/pnas.90.2.775","article-title":"Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African and American visceral leishmaniasis","volume":"90","author":"JM Burns","year":"1993","journal-title":"Proc Natl Acad Sci USA"},{"issue":"9","key":"pone.0319118.ref017","doi-asserted-by":"crossref","first-page":"594","DOI":"10.1093\/trstmh\/trv060","article-title":"Diagnostic accuracy of rK28-based immunochromatographic rapid diagnostic tests for visceral leishmaniasis: a prospective clinical cohort study in Sudan","volume":"109","author":"M Mukhtar","year":"2015","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"pone.0319118.ref018","doi-asserted-by":"crossref","first-page":"2454","DOI":"10.1128\/jcm.35.10.2454-2457.1997","article-title":"Evaluation of PCR for diagnosis of visceral leishmaniasis","volume":"35","author":"OF Osman","year":"1997","journal-title":"J Clin Microbiol"},{"issue":"9","key":"pone.0319118.ref019","doi-asserted-by":"crossref","first-page":"3147","DOI":"10.1128\/JCM.00372-14","article-title":"Molecular tools for diagnosis of visceral leishmaniasis: systematic review and meta-analysis of diagnostic test accuracy","volume":"52","author":"CM de Ruiter","year":"2014","journal-title":"J Clin Microbiol"},{"key":"pone.0319118.ref020","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/0014-4894(71)90098-1","article-title":"Trypanosoma cruzi: Agglutination activity of enzyme-treated epimastigotes","volume":"30","author":"NH Vattuone","year":"1971","journal-title":"Exp Parasitol"},{"issue":"2","key":"pone.0319118.ref021","doi-asserted-by":"crossref","first-page":"232","DOI":"10.4269\/ajtmh.1975.24.232","article-title":"A direct agglutination test for leishmaniasis","volume":"24","author":"DS Allain","year":"1975","journal-title":"Am J Trop Med Hyg"},{"key":"pone.0319118.ref022","first-page":"169","article-title":"A card agglutination test with stained trypanosomes (C.A.T.T.) for the serological diagnosis of T.B. gambiense trypanosomiasis","volume":"58","author":"E Magnus","year":"1978","journal-title":"Ann Soc Belg Med Trop"},{"issue":"4","key":"pone.0319118.ref023","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1016\/0035-9203(86)90149-5","article-title":"A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leishmaniasis","volume":"80","author":"AE Harith","year":"1986","journal-title":"Trans R Soc Trop Med Hyg"},{"issue":"7","key":"pone.0319118.ref024","doi-asserted-by":"crossref","first-page":"1321","DOI":"10.1128\/jcm.26.7.1321-1325.1988","article-title":"Improvement of a direct agglutination test for field studies of visceral leishmaniasis","volume":"26","author":"A el Harith","year":"1988","journal-title":"J Clin Microbiol"},{"issue":"8","key":"pone.0319118.ref025","doi-asserted-by":"crossref","first-page":"1984","DOI":"10.1128\/jcm.33.8.1984-1988.1995","article-title":"Evaluation of cleaving agents other than trypsin in direct agglutination test for further improving diagnosis of visceral leishmaniasis","volume":"33","author":"A el Harith","year":"1995","journal-title":"J Clin Microbiol"},{"issue":"12","key":"pone.0319118.ref026","doi-asserted-by":"crossref","first-page":"1731","DOI":"10.1099\/jmm.0.000858","article-title":"User and environment friendly direct agglutination test for the sero-diagnosis of visceral leishmaniasis: exclusion of formaldehyde and \u03b2-mercaptoethanol in test execution","volume":"67","author":"A Mahamoud","year":"2018","journal-title":"J Med Microbiol"},{"issue":"4","key":"pone.0319118.ref027","article-title":"Haematological malignancies as disorders negatively impacting specificities of the direct agglutination and rapid rK39 strip tests as reference diagnostics for visceral leishmanisis","volume":"5","author":"AE Nail","year":"2023","journal-title":"Access Microbiol"},{"issue":"4","key":"pone.0319118.ref028","doi-asserted-by":"crossref","first-page":"782","DOI":"10.4269\/ajtmh.19-0745","article-title":"Modifications in a Reference 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Harith","year":"2018","journal-title":"Open Forum Infect Dis"},{"issue":"4","key":"pone.0319118.ref034","doi-asserted-by":"crossref","first-page":"390","DOI":"10.4269\/ajtmh.1991.44.390","article-title":"Positive response to sodium antimony gluconate administration in visceral leishmaniasis seropositive patients","volume":"44","author":"S Chowdhury","year":"1991","journal-title":"Am J Trop Med Hyg"},{"issue":"8683","key":"pone.0319118.ref035","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/0140-6736(90)90313-T","article-title":"Outbreak of kala-azar in the Sudan","volume":"335","author":"P de Beer","year":"1990","journal-title":"Lancet"},{"issue":"3","key":"pone.0319118.ref036","doi-asserted-by":"crossref","first-page":"283","DOI":"10.4269\/ajtmh.1991.44.283","article-title":"A killing disease epidemic among displaced Sudanese population identified as visceral leishmaniasis","volume":"44","author":"P de Beer","year":"1991","journal-title":"Am J Trop Med Hyg"},{"issue":"11","key":"pone.0319118.ref037","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1046\/j.1360-2276.2003.01129.x","article-title":"Use of glycerol as an alternative to freeze-drying for long-term preservation of antigen for the direct agglutination test","volume":"8","author":"A el Harith","year":"2003","journal-title":"Trop Med Int Health"},{"key":"pone.0319118.ref038","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1099\/jmm.0.46753-0","article-title":"Evaluation of a glycerol-preserved antigen in the direct agglutination test for diagnosis of visceral leishmaniasis at rural level in eastern Sudan","volume":"55","author":"M el Mutasim","year":"2006","journal-title":"J Med Microbiol"},{"key":"pone.0319118.ref039","first-page":"15","article-title":"Application of direct agglutination test (DAT) for the diagnosis and sero-epidemiological studies of visceral leishmaniasis in Iran","volume":"1","author":"M Mohebali","year":"2006","journal-title":"Iran J Parasitol"},{"key":"pone.0319118.ref040","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/0035-9203(95)90487-5","article-title":"Visceral leishmaniasis in Bangladesh: the value of DAT as a diagnostic tool","volume":"89","author":"MA Al-Masum","year":"1995","journal-title":"Trans R Soc Trop Med Hyg"},{"issue":"5","key":"pone.0319118.ref041","doi-asserted-by":"crossref","first-page":"982","DOI":"10.4269\/ajtmh.15-0574","article-title":"Local Production of a Liquid Direct Agglutination Test as a Sustainable Measure for Control of Visceral Leishmaniasis in Sudan","volume":"94","author":"HA Osman","year":"2016","journal-title":"Am J Trop Med Hyg"},{"issue":"1","key":"pone.0319118.ref042","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1186\/s12866-020-01780-0","article-title":"Validation of in-house liquid direct agglutination test antigen: the potential diagnostic test in visceral Leishimaniasis endemic areas of Northwest Ethiopia","volume":"20","author":"B Ayelign","year":"2020","journal-title":"BMC Microbiol"},{"key":"pone.0319118.ref043","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1099\/jmm.0.063891-0","article-title":"Performance of an indigenous \u03b2-mercaptoethanol-modified antigen in comparison with a commercial reference in direct agglutination test for detection of canine visceral leishmaniasis","volume":"63","author":"SJ Semi\u00e3o-Santos","year":"2014","journal-title":"J Med Microbiol"},{"key":"pone.0319118.ref044","first-page":"279","article-title":"Visceral Leishmaniasis in Iran: An Update on Epidemiological Features from 2013 to 2022","volume":"18","author":"M Mohebali","year":"2023","journal-title":"Iran J Parasitol"},{"issue":"6","key":"pone.0319118.ref045","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1007\/BF00931580","article-title":"Prevalence of agglutinating anti-Leishmania antibodies in two multi-thousand Bengoli communities","volume":"79","author":"MS Chowdhury","year":"1993","journal-title":"Parasitol Res"},{"issue":"7","key":"pone.0319118.ref046","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1080\/00034983.1999.11813475","article-title":"Evaluation of a standardized direct agglutination test (DAT) for the diagnosis of visceral leishmaniasis in Kenya","volume":"93","author":"PA Mbati","year":"1999","journal-title":"Ann Trop Med Parasitol"},{"issue":"5","key":"pone.0319118.ref047","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1128\/CDLI.5.5.717-720.1998","article-title":"rK39 enzyme-linked immunosorbent assay for diagnosis of Leishmania donovani infection","volume":"5","author":"EE Zijlstra","year":"1998","journal-title":"Clin Diagn Lab Immunol"},{"issue":"2","key":"pone.0319118.ref048","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1128\/CVI.00473-10","article-title":"Persistence of Leishmania donovani antibodies in past visceral leishmaniasis cases in India","volume":"18","author":"K Gidwani","year":"2011","journal-title":"Clin Vaccine Immunol"},{"issue":"10","key":"pone.0319118.ref049","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pntd.0003273","article-title":"IgG1 as a potential biomarker of post-chemotherapeutic relapse in visceral leishmaniasis, and adaptation to a rapid diagnostic test","volume":"8","author":"T Bhattacharyya","year":"2014","journal-title":"PLoS Negl Trop Dis"},{"key":"pone.0319118.ref050","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.actatropica.2017.11.010","article-title":"Identification of an area predominantly endemic for childhood and adolescent visceral leishmaniasis in central Sudan","volume":"178","author":"A Mahamoud","year":"2018","journal-title":"Acta Trop"},{"issue":"1","key":"pone.0319118.ref051","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.trstmh.2007.09.003","article-title":"Diagnostic tests for kala-azar: a multi-centre study of the freeze-dried DAT, rK39 strip test and KAtex in East Africa and the Indian subcontinent","volume":"102","author":"M Boelaert","year":"2008","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"pone.0319118.ref052","doi-asserted-by":"crossref","first-page":"76","DOI":"10.4269\/ajtmh.2006.74.76","article-title":"Evaluation of a new recombinant K39 rapid diagnostic for Sudanese visceral leishmaniasis","volume":"74","author":"K Ritmeir","year":"2006","journal-title":"Am J Trop Med Hyg"},{"key":"pone.0319118.ref053","volume-title":"WHO guidelines for the treatment of visceral leishmaniasis in HIV co-infected patients in East Africa and South-East Asia","year":"2022"},{"key":"pone.0319118.ref054","article-title":"Evaluation of rapid diagnostic tests: visceral leishmaniasis","author":"M Boelaert","year":"2007","journal-title":"Nat Rev Microbiol"},{"issue":"6","key":"pone.0319118.ref055","doi-asserted-by":"crossref","first-page":"929","DOI":"10.4269\/ajtmh.2009.80.929","article-title":"Field evaluation of rK39 test and direct agglutination test for diagnosis of visceral leishmaniasis in a population with high prevalence of human immunodeficiency virus in Ethiopia","volume":"80","author":"R ter Horst","year":"2009","journal-title":"Am J Trop Med Hyg"},{"issue":"1","key":"pone.0319118.ref056","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1179\/000349802125000475","article-title":"The performance of direct agglutination tests (DAT) in the diagnosis of visceral leishmaniasis among Ethiopian patients with HIV co-infection","volume":"96","author":"A Hailu","year":"2002","journal-title":"Ann Trop Med Parasitol"},{"issue":"6","key":"pone.0319118.ref057","doi-asserted-by":"crossref","first-page":"526","DOI":"10.1007\/BF00931059","article-title":"Introduction of an improved direct agglutination test for the detection of Leishmania infantum infection in southern France","volume":"76","author":"PM de Korte","year":"1990","journal-title":"Parasitol Res"},{"issue":"2","key":"pone.0319118.ref058","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.tmaid.2013.09.001","article-title":"Emergence of co-infection of visceral leishmaniasis in HIV-positive patients in northeast Iran: a preliminary study","volume":"12","author":"R Shafiei","year":"2014","journal-title":"Travel Med Infect Dis"},{"issue":"3","key":"pone.0319118.ref059","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0116408","article-title":"Heterogeneity of Leishmania donovani parasites complicates diagnosis of visceral leishmaniasis: comparison of different serological tests in three endemic regions","volume":"10","author":"E Abass","year":"2015","journal-title":"PLoS One"},{"key":"pone.0319118.ref060","article-title":"Strong association between serological status and probability of progression to clinical visceral Leishmaniasis in prospective cohort studies in India and Nepal","author":"E Hasker","year":"2014","journal-title":"PLoS Negl Trop Dis"},{"issue":"1","key":"pone.0319118.ref061","doi-asserted-by":"crossref","first-page":"10426","DOI":"10.1038\/s41598-022-14490-6","article-title":"Validation of a mixture of rK26 and rK39 antigens from Iranian strain of Leishmania infantum to detect anti-Leishmania antibodies in human and reservoir hosts","volume":"12","author":"BR Hosseini Farash","year":"2022","journal-title":"Sci Rep"},{"issue":"3","key":"pone.0319118.ref062","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pntd.0007216","article-title":"Determinants for progression from asymptomatic infection to symptomatic visceral leishmaniasis: A cohort study","volume":"13","author":"J Chakravarty","year":"2019","journal-title":"PLoS Negl Trop Dis"},{"issue":"4","key":"pone.0319118.ref063","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1111\/j.1365-3156.2009.02242.x","article-title":"PCR and direct agglutination as Leishmania infection markers among healthy Nepalese subjects living in areas endemic for Kala-Azar","volume":"14","author":"NR Bhattarai","year":"2009","journal-title":"Trop Med Int Health"},{"key":"pone.0319118.ref064","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/S1995-7645(12)60008-X","article-title":"Preparation and evaluation of a glycerol-preserved direct agglutination antigen for long-term preservation: a comparative study of the detection of anti-Leishmania infantum antibodies in human and dog","author":"B Akhoundi","year":"2012","journal-title":"Asian Pac J Trop Med"},{"issue":"7","key":"pone.0319118.ref065","doi-asserted-by":"crossref","first-page":"1742","DOI":"10.1128\/jcm.33.7.1742-1745.1995","article-title":"Leish-KIT, a stable direct agglutination test based on freeze-dried antigen for serodiagnosis of visceral leishmaniasis","volume":"33","author":"SE Meredith","year":"1995","journal-title":"J Clin 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