Abstract
In this study, serological and molecular biological approaches were implicated to be used in the diagnosis of leptospirosis. We developed specific antigens of leptospira by using leptospiral outer membrane, LipL32, and leptospiral immunoglobulin-like protein A (LigA). By using these recombinant antigens, ELISA for IgG and IgM tests for sera from leptospirosis cases found in Sri Lanka were performed. These cases were already diagnosed by the microscopic agglutination test (MAT) and PCR. Serological approaches revealed the preparation of specific antigens to be used in ELISA system and the sensitivity of ELISA system compared with MAT. The chi-square test (χ 2) defined that IgG-LipL32, IgG-LigA, and IgM-ELISA in comparison with MAT were sensitive as 5.439 (significance p ≤ 0.01), 9.188 (significance p ≤ 0.001), and 5.915 (significance p ≤ 0.01), respectively. Molecular studies approaches revealed the sensitivity of PCR to be diagnostic tool in leptospirosis. PCR can be considered as effective diagnostic tools to diagnose leptospirosis. From the results of experiments for comparing sensitivity, several contradictions were found. Too much amount of DNA might be inhibiting specific PCR. Anyway, both PCR methods targeting flaB gene and WHO recommendations were followed to detect genomic DNA of leptospira effectively.
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Acknowledgments
This work was supported in part by Global COE Program “Establishment of International Collaboration Centers for Zoonosis Control,” MEXT, Japan. We are grateful to all members of Prof. Jiro Arikawa’s laboratory, Department of Microbiology, Graduate School of Medicine, Hokkaido University, for their kindness and supervision and for providing the place and equipments for this training course’s experiments. We also thank Dr. Nobuo Koizumi and Dr. Chandika D. Gamage for providing the whole genome of Leptospira and serum samples.
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Widiasih, D.A., Isozumi, R., Yoshimatsu, K., Arikawa, J. (2017). Serological and Molecular Biological Diagnosis for Leptospirosis. In: Isnansetyo, A., Nuringtyas, T. (eds) Proceeding of the 1st International Conference on Tropical Agriculture. Springer, Cham. https://doi.org/10.1007/978-3-319-60363-6_56
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DOI: https://doi.org/10.1007/978-3-319-60363-6_56
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