Development of In-House ELISA using recombinant LipL32 for Detection of Human Leptospirosis in Indonesia

Sumarningsih Sumarningsih; Gita Sekarmila; Andi Mulyadi; Ahpas Ahpas; Simson Tarigan

doi:10.22146/jsv.90085

Development of In-House ELISA using recombinant LipL32 for Detection of Human Leptospirosis in Indonesia

https://doi.org/10.22146/jsv.90085

Sumarningsih Sumarningsih^(1*), Gita Sekarmila⁽²⁾, Andi Mulyadi⁽³⁾, Ahpas Ahpas⁽⁴⁾, Simson Tarigan⁽⁵⁾

(1) Research Center for Veterinary Science, National research and innovation agency (BRIN)
(2) Research Center for Veterinary Science, National research and innovation agency (BRIN)
(3) Research Center for Veterinary Science, National research and innovation agency (BRIN)
(4) Research Center for Veterinary Science, National research and innovation agency (BRIN)
(5) Research Center for Veterinary Science, National research and innovation agency (BRIN)
(*) Corresponding Author

Abstract

Early laboratory confirmation is important for the accurate diagnosis and treatment of patient infected by leptospirosis. However, Microscopic agglutination test (MAT) as the gold standard for detection of human leptospirosis has many limitation and only available in reference laboratories. Therefore, many studies suggested LipL32 protein as a good candidate for development of leptospirosis detection kit because it is highly conserved and produced only in pathogenic Leptospira species. In this study, we aim to investigate the performance of our in-house ELISA using recombinant LipL32 to detect leptospirosis in Indonesia. Fourteen human sera were used in this study and the infection status were determine using MAT. The result showed that nine of eleven MAT positive sera were successfully recognized by LipL32 ELISA. The antibody binding to LipL32 was also confirm by immunoblot. There was one of three MAT negative sera has high OD above 0.5 in ELISA, but it showed negative reaction in immunoblot result. Overall, this study demonstrated that recombinant LipL32 protein can recognized antibody from human leptospirosis and can be used as a universal antigen to detect infection by any serovars of pathogenic leptospira.

Keywords

ELISA; human; leptospirosis; LipL32

Full Text:

PDF

References

Bomfim, M.R.Q., Ko, A., Koury, M.C. (2005). Evaluation of the recombinant LipL32 in enzyme-linked immunosorbent assay for the serodiagnosis of bovine leptospirosis. Vet. Microbiol. 109, 89–94. https://doi.org/10.1016/j.vetmic.2005.05.002. Costa, F., Hagan, J.E., Calcagno, J., Kane, M., Torgerson, P., Martinez-Silveira, M.S., Stein, C., Abela-Ridder, B., Ko, A.I. (2015). Global Morbidity and Mortality of Leptospirosis: A Systematic Review. PLoS Negl. Trop. Dis. 9, 1–19. https://doi.org/10.1371/journal.pntd.0003898. Gasem, M.H., Hadi, U., Alisjahbana, B., Tjitra, E., Hapsari, M.M.D.E.A.H., Lestari, E.S., Aman, A.T., Lokida, D., Salim, G., Kosasih, H., Merati, K.T.P., Laras, K., Arif, M., Lukman, N., Sudarmono, P., Lisdawati, V., Lau, C.-Y., Neal, A., Karyana, M. (2020). Leptospirosis in Indonesia: diagnostic challenges associated with atypical clinical manifestations and limited laboratory capacity. BMC Infect. Dis. 20, 179. https://doi.org/10.1186/s12879-020-4903-5. Goris, M.G.A., Leeflang, M.M.G., Loden, M., Wagenaar, J.F.P., Klatser, P.R., Hartskeerl, R.A., Boer, K.R. (2013). Prospective Evaluation of Three Rapid Diagnostic Tests for Diagnosis of Human Leptospirosis. PLoS Negl. Trop. Dis. 7, 1–11. https://doi.org/10.1371/journal.pntd.0002290. Haake, D.A., Chao, G., Zuerner, R.L., Barnett, J.K., Barnett, D., Mazel, M., Matsunaga, J., Levett, P.N., Bolin, C.A. (2000). The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection. Infect. Immun. 68, 2276–2285. https://doi.org/10.1128/IAI.68.4.2276-2285.2000. Levett, P.N. (2001). Leptospirosis. Clin. Microbiol. Rev. 14, 296–326. https://doi.org/10.1128/CMR.14.2.296-326.2001. Limmathurotsakul, D., Turner, E.L., Wuthiekanun, V., Thaipadungpanit, J., Suputtamongkol, Y., Chierakul, W., Smythe, L.D., Day, N.P.J., Cooper, B., Peacock, S.J. (2012). Fool’s gold: Why imperfect reference tests are undermining the evaluation of novel diagnostics: a reevaluation of 5 diagnostic tests for leptospirosis. Clin. Infect. Dis. an Off. Publ. Infect. Dis. Soc. Am. 55, 322–331. https://doi.org/10.1093/cid/cis403. Musso, D., La Scola, B. (2013). Laboratory diagnosis of leptospirosis: A challenge. J. Microbiol. Immunol. Infect. 46, 245–252. https://doi.org/https://doi.org/10.1016/j.jmii.2013.03.001. Nurhidayat, D. (2023). Kemenkes Keluarkan Surat Edaran Penanganan Leptospirosis untuk Dinkes Daerah Access from: https://mediaindonesia.com/humaniora/565146/kemenkes-keluarkan-surat-edaran-penanganan-leptospirosis-untuk-dinkes-daerah. Access on 22 October 2023. Picardeau, M. (2013). Diagnosis and epidemiology of leptospirosis. Med. Mal. Infect. 43, 1–9. https://doi.org/10.1016/j.medmal.2012.11.005. Sumarningsih, Susanti, Tarigan, S. (2017). Evaluation of LipL32 ELISA for detection of bovine leptospirosis in West Java. J. Ilmu Ternak dan Vet. 80–90. Waritani, T., Chang, J., McKinney, B., & Terato, K. (2017). An ELISA protocol to improve the accuracy and reliability of serological antibody assays. MethodsX, 4, 153–165. https://doi.org/https://doi.org/10.1016/j.mex.2017.03.002.

DOI: https://doi.org/10.22146/jsv.90085