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Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters

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 diff...

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Autores principales: Hamer, Micaela, Watanabe, Olivia, Saraullo, Vanina Rosa, Ortega, Facundo Miguel, Sanchez, Maria Cristina, Martinez, Mara Leila, Brihuega, Bibiana Felicitas, Grune Loffler, Sylvia
Formato: info:ar-repo/semantics/artículo
Lenguaje:Inglés
Publicado: Springer 2023
Materias:
Animal Models
Infectious Diseases
Leptospira
PCR
Hamsters
Modelos Animales
Enfermedades Infecciosas
Reacción en Cadena de la Polimerasa
Hámster
Loop-mediated Isothermal Amplification
Sensitivity
Specificity
Amplificación Isotérmica Mediada por Bucle
Sensibilidad
Especificidad
Acceso en línea:http://hdl.handle.net/20.500.12123/15227
https://link.springer.com/article/10.1007/s11259-023-10183-1
https://doi.org/10.1007/s11259-023-10183-1
Sumario: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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