Surveillance of leptospiral reservoirs in synanthropic rodents using loop-mediated isothermal amplification
Micaela Hamer1, Vanina Saraullo1, Emiliano Muschetto2
1Laboratorio de Leptospirosis. Instituto de Patobiología - IPVET (UEDD INTA-CONICET), Centro de Investigación en Ciencias Veterinarias y Agronómicas (CICVYA), Instituto Nacional de Tecnología Agropecuaria (INTA), Nicolás Repetto y de los Reseros s/n, Hurlingham, Buenos Aires, Argentina.
Abstract:
Leptospirosis, a zoonotic disease caused by pathogenic Leptospira spp., represents a major public health concern due to its impact on both rural and urban populations. Rodents, particularly Rattus norvegicus, Rattus rattus, and Mus musculus, are key reservoirs, excreting leptospires in their urine and contributing to environmental contamination. In this study, we evaluated the efficacy of loop-mediated isothermal amplification (LAMP), a molecular diagnostic tool, for detecting leptospiral DNA in kidney samples from captured rodents. LAMP results were compared with the standard lipL32 PCR assay. Leptospiral DNA was detected in 9.0 % (14/156) of samples, with 5.8 % positive by both LAMP and lipL32 PCR and 3.2 % positive by LAMP alone. No samples were positive by PCR and negative by LAMP. Cohen's Kappa index (0.77) indicated substantial agreement between the two methods. The higher sensitivity of LAMP, its ability to detect both pathogenic and intermediate leptospiral strains, and its cost-effectiveness make it a valuable tool for low-resource settings. However, the technique's inability to differentiate between Leptospira species highlights the need for complementary methods for epidemiological studies. These findings contribute to the understanding of rodent leptospirosis reservoirs and offer practical diagnostic solutions for veterinary and public health surveillance.
Insights
Loop-mediated isothermal amplification (LAMP) effectively detects leptospiral DNA in rodent kidneys, showing substantial agreement with PCR. This cost-effective method aids leptospirosis surveillance in resource-limited settings.
Area of Science:
- Veterinary Medicine
- Molecular Diagnostics
- Zoonotic Diseases
Background:
- Leptospirosis is a significant zoonotic disease with global public health implications.
- Rodents are primary reservoirs for pathogenic Leptospira spp., facilitating environmental contamination.
- Accurate diagnostics are crucial for understanding and controlling leptospirosis transmission.
Purpose of the Study:
- To evaluate the diagnostic efficacy of loop-mediated isothermal amplification (LAMP) for detecting leptospiral DNA in rodent kidney samples.
- To compare LAMP performance against the established lipL32 PCR assay.
- To assess LAMP's utility as a diagnostic tool for leptospirosis surveillance.
Main Methods:
- Rodent kidney samples were analyzed using both loop-mediated isothermal amplification (LAMP) and lipL32 PCR.
- Leptospiral DNA detection rates were compared between the two molecular diagnostic methods.
- Statistical analysis, including Cohen's Kappa index, was used to determine agreement.
Main Results:
- Leptospiral DNA was detected in 9.0% of rodent samples.
- LAMP demonstrated higher sensitivity, detecting 3.2% more positive samples than PCR alone.
- A substantial agreement (Cohen's Kappa = 0.77) was observed between LAMP and lipL32 PCR methods.
Conclusions:
- LAMP is a sensitive, cost-effective molecular diagnostic tool for detecting leptospiral DNA in rodents.
- The method shows promise for leptospirosis surveillance, particularly in low-resource settings.
- Further studies incorporating species identification are recommended for comprehensive epidemiological analysis.
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