Abstract
Leptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with different colorimetric indicators. LAMP was able to detect DNA from pathogenic and intermediate leptospires, while non-pathogenic leptospires and other non-leptospiral microorganisms were negative. LAMP assay combined with calcein showed a tenfold higher limit of detection (1 ng of leptospiral DNA per reaction) than LAMP combined with hydroxynaphthol blue or end-point PCR lipL32 (10 ng of DNA per reaction). Animal samples were collected from infected and non-infected Golden Syrian hamsters (Mesocricetus auratus) to evaluate and compare the performance of LAMP and PCR. These techniques showed a substantial agreement according to Cohen’s kappa statistic, being both useful techniques for detecting leptospiral DNA in clinical samples. Overall, this study demonstrates that the LAMP assay is a sensitive, specific, rapid, and simple tool for the detection of leptospiral DNA. It has the potential to facilitate the diagnosis of leptospirosis, particularly in low-income regions with limited diagnosis resources.
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Acknowledgements
We thank Ph.D. Pablo Chacana, Lic. Matias Ricchetta and Ph.D. Karina Trono for their support in laboratory animal management, and SENASA for providing hamsters and virulent pathogenic Leptospira strain.
Funding
This research received financial support from Fund for Scientific and Technological Research (FONCyT) of the National Agency for the Promotion of Research, Technological Development and Innovation (PICT Start-Up 2016–4815). Bioch. Micaela Hamer has a PhD Scholarship financed by the National Research Council of Argentina (CONICET).
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Micaela Hamer, Olivia Watanabe, Vanina Saraullo, Facundo Ortega and Cristina Sánchez. Mara Martínez, Bibiana Brihuega, and Sylvia Grune Loffler assisted with discussion. The first draft of the manuscript was written by Micaela Hamer and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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All animal procedures were approved as standard operating procedure (SOP) #48/18, by Institutional Committee for the Care and Use of Experimental Animals (CICUAE), from National Institute of Agricultural Technology (INTA).
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Hamer, M., Watanabe, O., Saraullo, V. et al. Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters. Vet Res Commun 48, 103–111 (2024). https://doi.org/10.1007/s11259-023-10183-1
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DOI: https://doi.org/10.1007/s11259-023-10183-1