Abstract
Cutaneous leishmaniasis (CL) in Sri Lanka is caused by Leishmania donovani, a parasite widely known to cause visceral leishmaniasis. Despite the fact that CL is not generally believed to elicit serological immune responses, recent studies show the presence of antibody responses against this atypical form of CL. This study assesses the potential of using recombinant K39 (rK39), KMP11, and crude parasite antigen-based indirect ELISAs as serological diagnostic tools and measures of exposure for CL in Sri Lanka. The study used serum samples from confirmed CL patients (n = 266) and apparently healthy individuals from endemic settings (n = 411). Serum samples from individuals residing in non-endemic areas were used as negative controls. In-house indirect ELISAs were optimized and validated for recombinant antigens. Previously validated crude parasite extract-based indirect ELISA was performed for comparison. The statistical analyses were performed using SPSS v26.0. The rK39 (sensitivity = 71.2%, specificity = 64%) and KMP11 (sensitivity = 79.2%, specificity = 71.4%) based indirect ELISA were shown to be less suitable for the diagnosis of CL, while crude parasite extract-based indirect ELISA (sensitivity = 82.4%, specificity = 85.7%) might be a better method of diagnosis. All 03 ELISAs seemed to be good methods as measures of exposure since correlations were observed between the seropositivity of all 03 ELISAs (rK39: p = 0.037, KMP11: p = 0.007, CrudeAg: p = 0.000) with provincial case incidences. The findings will be important in identifying the disease hotspots in order to design the control measures for CL induced by L. donovani in Sri Lanka.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
The datasets generated and analyzed during the current study are available from the corresponding author on a reasonable request.
References
Athukorale DN, Seneviratne JKK, Ihalamulla RL, Premaratne UN (1992) Locally acquired cutaneous leishmaniasis in Sri Lanka. J Trop Med Hyg 95(6):432–433
Basu R, Bhaumik S, Basu JM, Naskar K, De T, Roy S (2005) Kinetoplastid membrane protein-11 DNA vaccination induces complete protection against both pentavalent antimonial-sensitive and -resistant strains of Leishmania donovani that correlates with inducible nitric oxide synthase activity and IL-4 generation: evidence for mixed Th1- and Th2-like responses in visceral leishmaniasis. J Immunol 174(11):7160–7171. https://doi.org/10.4049/JIMMUNOL.174.11.7160
Carrillo E, Moreno J (2019) Editorial: Biomarkers in leishmaniasis. Front Cell Infect Microbiol 9(November):1–4. https://doi.org/10.3389/fcimb.2019.00388
Castellani A (1904) “Leishmania Donovani” in Ceylon. Seventy-Second Annual Meeting of the British Medical Association, BMJ2 629–78. https://doi.org/10.1136/bmj.2.2281.629
CDC (2020) Leishmaniasis. https://www.cdc.gov/parasites/leishmaniasis/index.html. Accessed 21 February 2023
Conde L et al (2022) Humoral response in Leishmaniasis. Front Cell Infect Microbiol 12:1797. https://doi.org/10.3389/FCIMB.2022.1063291/BIBTEX
de Mendonça SCF, Cysne-Finkelstein L, de Matos DCS (2015) Kinetoplastid membrane protein-11 as a vaccine candidate and a virulence factor in Leishmania. Front Immunol 6:524. https://doi.org/10.3389/fimmu.2015.00524
De Silva G et al (2017) Efficacy of a new rapid diagnostic test kit to diagnose Sri Lankan cutaneous leishmaniasis caused by Leishmania donovani. PLoS ONE 12(11):1–14. https://doi.org/10.1371/journal.pone.0187024
De Silva NL (2022) Validation of an in-house ELISA method in the diagnosis of cutaneous leishmaniasis caused by Leishmania donovani in Hambantota District, Sri Lanka. Microorganisms 10(5). https://doi.org/10.3390/microorganisms10050921
Deepachandi B et al (2020) First Serological Study Revealing High Humoral Response and Evidence for Antigenic Heterogeneity in Leishmania donovani Induced CL in Sri Lanka. Biomed Res Int 2020:5271657. https://doi.org/10.1155/2020/5271657
Drakeley CJ et al (2005) Estimating medium- and long-term trends in malaria transmission by using serological markers of malaria exposure. Proc Natl Acad Sci USA 102(14):5108–5113. https://doi.org/10.1073/pnas.0408725102
Epidemiology Unit (2019) Leishmaniasis. https://www.epid.gov.lk/search?title=leishmaniasis Accessed 30 May 2023
Farahmand M, Nahrevanian H (2016) Application of recombinant proteins for serodiagnosis of visceral leishmaniasis in humans and dogs. Iran Biomed J 20(3):128–134. https://doi.org/10.7508/ibj.2016.03.001
Greenhouse B et al (2018) Taking sharper pictures of malaria with CAMERAs: combined antibodies to measure exposure recency assays. Am J Trop Med Hyg 99(5):1120–1127. https://doi.org/10.4269/AJTMH.18-0303
Guimaraes MCS, Celeste BJ, Franco EL (1990) Diagnostic performance indices for immunofluorescent tests and enzyme immunoassays of leishmaniasis sera from northern and north-eastern Brazil. Bull World Health Organ 68(1):39–43
Karunaweera ND (2009) Leishmania donovani causing cutaneous leishmaniasis in Sri Lanka: a wolf in sheep’s clothing? Trends Parasitol 25(10):458–463. https://doi.org/10.1016/J.PT.2009.07.002
Karunaweera ND et al (2020) Spatial epidemiologic trends and hotspots of leishmaniasis, Sri Lanka, 2001–2018. Emerg Infect Dis 26(1):1. https://doi.org/10.3201/EID2601.190971
Karunaweera ND et al (2021) Spatiotemporal distribution of cutaneous leishmaniasis in Sri Lanka and future case burden estimates. PLoS Negl Trop Dis 15(4):e0009346. https://doi.org/10.1371/JOURNAL.PNTD.0009346
Kaushal H, Bhattacharya SK, Verma S, Salotra P (2017) Serological and molecular analysis of leishmania infection in healthy individuals from two districts of West Bengal, India, endemic for visceral leishmaniasis. Am J Trop Med Hyg 96(6):1448–1455. https://doi.org/10.4269/AJTMH.16-0592
Kumar R, Pai K, PathakK SS (2001) Enzyme-linked immunosorbent assay for recombinant K39 antigen in diagnosis and prognosis of Indian visceral leishmaniasis. Clin Diagn Lab Immunol 8(6):1220–1224. https://doi.org/10.1128/CDLI.8.6.1220-1224.2001/ASSET/83DC25B1-1175-4A9A-BC30-608D577FE7D1/ASSETS/GRAPHIC/CD0610055002.JPEG
Lauricella MA et al (2016) An rK28-based immunoenzymatic assay for the diagnosis of canine visceral leishmaniasis in Latin America. Am J Trop Med Hyg 95(1):92–98. https://doi.org/10.4269/ajtmh.13-0768
Maia Z et al (2012) Comparative study of rK39 leishmania antigen for serodiagnosis of visceral leishmaniasis: systematic review with meta-analysis. PLoS Negl Trop Dis. https://doi.org/10.1371/journal.pntd.0001484
Martínez-Moreno A, Moreno T, Martínez-Moreno FJ, Acosta I, Hernández S (1995) Humoral and cell-mediated immunity in natural and experimental canine leishmaniasis. Vet Immunol Immunopathol 48(3–4):209–220. https://doi.org/10.1016/0165-2427(95)05434-8
Mcgwire BS, Satoskar AR (2014) Leishmaniasis: clinical syndromes and treatment. QJM 107(1):7–14. https://doi.org/10.1093/qjmed/hct116
Michel G, Pomares C, Ferrua B, Marty P (2011) Importance of worldwide asymptomatic carriers of Leishmania infantum (L. chagasi) in human. Acta Tropica 119(2–3):69–75. https://doi.org/10.1016/J.ACTATROPICA.2011.05.012
Passos S et al (2005) Recombinant Leishmania antigens for serodiagnosis of visceral leishmaniasis. Clin Diagn Lab Immunol 12(10):1164–1167. https://doi.org/10.1128/CDLI.12.10.1164-1167.2005
Piyasiri SB, Samaranayake TN, Silva H, Manamperi NH, Karunaweera ND (2022) ELISA-based evaluation of antibody response to Leishmania in a region endemic for cutaneous leishmaniasis. Parasite Immunol 44(9):e12940. https://doi.org/10.1111/PIM.12940
Rezaei Z et al (2019) Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis. Parasit Vectors 12(1):1–9. https://doi.org/10.1186/S13071-019-3839-3/FIGURES/4
Silva H, Liyanage A, Deerasinghe T, Chandrasekara V, Chellappan K, Karunaweera ND (2021) Treatment failure to sodium stibogluconate in cutaneous leishmaniasis: a challenge to infection control and disease elimination. PLoS ONE 16(10):1–16. https://doi.org/10.1371/journal.pone.0259009. (October)
Singh OP, Hasker E, Sacks D, Boelaert M, Sundar S (2014) Asymptomatic leishmania infection: a new challenge for leishmania control. Clin Infect Dis 58(10):1424–1429. https://doi.org/10.1093/CID/CIU102
Singh S, Kumari V, Singh N (2002) Predicting kala-azar disease manifestations in asymptomatic patients with latent Leishmania donovani infection by detection of antibody against recombinant K39 antigen. Clin Diagn Lab Immunol 9(3):568–572. https://doi.org/10.1128/CDLI.9.3.568-572.2002
Siriwardana H, Chandrawansa P, Sirimanna G, Karunaweera N (2012) Leishmaniasis in Sri Lanka: a decade old story. Sri Lankan J Infect Dis 2(2):2. https://doi.org/10.4038/sljid.v2i2.4420
Siriwardana YD, Deepachandi B, Ranasinghe S, Soysa P, Karunaweera N (2018) Evidence for seroprevalence in human localized cutaneous leishmaniasis caused by Leishmania donovani in Sri Lanka. BioMed Res Int 2018. https://doi.org/10.1155/2018/9320367
Siriwardana Y et al (2019) Trends in recently emerged Leishmania donovani induced cutaneous leishmaniasis, Sri Lanka, for the first 13 years. BioMed Res Int 2019. https://doi.org/10.1155/2019/4093603
Srivastava P et al (2013) Molecular and serological markers of Leishmania donovani infection in healthy individuals from endemic areas of Bihar. India Trop Med Int Health 18(5):548–554. https://doi.org/10.1111/TMI.12085
Tan XT, Amran F, Cheong KC, Ahmad N (2014) In-house ELISA screening using a locally-isolated Leptospira in Malaysia: determination of its cut-off points. BMC Infect Dis 14(1):1–5. https://doi.org/10.1186/S12879-014-0563-7/TABLES/2
Verhey KJ, Kaul N, Soppina V (2011) Kinesin assembly and movement in cells. Annu Rev Biophys 40(1):267–288. https://doi.org/10.1146/ANNUREV-BIOPHYS-042910-155310
WHO (2023) Leishmaniasis. https://www.who.int/news-room/fact-sheets/detail/leishmaniasis Accessed 11 April 2023
Zeyrek FY, Korkmaz M, Özbel Y (2007) Serodiagnosis of anthroponotic cutaneous leishmaniasis (ACL) caused by Leishmania tropica in Sanliurfa Province, Turkey, where ACL is highly endemic. Clin Vaccine Immunol 14(11):1409. https://doi.org/10.1128/CVI.00133-07
Zijlstra EE et al (1998) rK39 enzyme-linked immunosorbent assay for diagnosis of Leishmania donovani infection. Clin Diagn Lab Immunol 5(5):717–720. https://doi.org/10.1128/cdli.5.5.717-720.1998
Funding
This study was funded by the National Institute of Allergy and Infectious Diseases of the National Institute of Health, USA, under award number U01AI136033.
Author information
Authors and Affiliations
Contributions
Conceptualization: N.K., R.D.; methodology: R.D.; investigation and sample collection: N.A., S.B.P., N.S., H.S., N.M.; lab work and validation: N.A., I.W., N.C., S.B.P., C.K.; statistical analysis: C.K., R.D.; writing—original draft preparation: C.K., I.W.; writing—review and editing: N.A., C.K., R.D., S.B.P., N.S., H.S., N.K.; funding acquisition: N.K.; resources: N.K.; supervision: N.K., R.D.
Corresponding author
Ethics declarations
Ethics approval
The study was performed in accordance with the ethics standards as laid down in the 1964 Declaration of Helsinki and its later amendments. Ethics approval for the study was obtained from the Ethics Review Committee of the Faculty of Medicine, University of Colombo, Sri Lanka (EC-17–062).
Consent to participate
Written informed consent was obtained from all the study participants.
Competing interests
The authors declare no competing interests.
Additional information
Section Editor: Nawal Hijjawi
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Charani Karunathilake and Narmadha Alles are joint first authors.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Karunathilake, C., Alles, N., Dewasurendra, R. et al. The use of recombinant K39, KMP11, and crude antigen-based indirect ELISA as a serological diagnostic tool and a measure of exposure for cutaneous leishmaniasis in Sri Lanka. Parasitol Res 123, 77 (2024). https://doi.org/10.1007/s00436-023-08103-y
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00436-023-08103-y