Novel Target for Pathogenic Leptospira Detection via Colorimetric Loop-Mediated Isothermal Amplification (LAMP): Development, Validation, and Comparative Analysis.
4 affiliations
- 1Center for Research Innovation and Biomedical Informatics, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand.
- 2Department of Medical Technology, Faculty of Pharmacy, University of Santo Tomas, Manila, Philippines.
- 3The Graduate School, University of Santo Tomas, Manila, Philippines.
- 4School of Health Sciences, University of Santo Tomas, General Santo, Philippines.
Abstract
Background
Leptospirosis is a globally prevalent zoonotic disease that remains challenging to diagnose due to its nonspecific symptoms and the limitations of current diagnostic methods and targets. To address these challenges, we developed and validated a naked-eye loop-mediated isothermal amplification (LAMP) assay targeting the newly identified orpH region for the rapid detection of pathogenic Leptospira species.Methods
Comparative genomic analysis identified orpH as a conserved sequence among pathogenic Leptospira. A colorimetric LAMP assay (LAMP-orpH) was developed and evaluated using reference strains and serum samples obtained from patients with suspected leptospirosis. The diagnostic performance of the assay on these serum samples was evaluated in comparison with conventional PCR and the microscopic agglutination test (MAT), with the real-time PCR assay targeting lipL32 designated as the reference standard.Results
The LAMP-orpH assay demonstrated 87.2% sensitivity and 94.8% specificity among 76 suspected cases and 29 controls, showing strong agreement with real-time PCR (κ = 0.826). It outperformed MAT (63.8% sensitivity) and conventional PCR (23.4% sensitivity), achieving a detection limit of 10² cells/mL in spiked serum.Conclusion
The LAMP-orpH assay offers a rapid, sensitive, and specific tool for early leptospirosis diagnosis. Its simple colorimetric readout and minimal equipment requirements make it ideal for use in rural and resource-limited settings, providing a practical alternative to MAT, conventional PCR, and qPCR.
Related
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- Development of a novel loop-mediated isothermal amplification (LAMP) assay for Balamuthia mandrillaris
- Development of Loop-Mediated Isothermal Amplification Assays for Rapid and Sensitive Detection of Three Viruses Infecting Areca catechu L.
BibTeX
@article{ghoraishizadeh2026novel,
title = {{Novel Target for Pathogenic Leptospira Detection via Colorimetric Loop-Mediated Isothermal Amplification (LAMP): Development, Validation, and Comparative Analysis.}},
author = {Peyman Ghoraishizadeh and Benjie Clemente and Kween Saimuang and Sakaorat Lertjuthaporn and Sukhonta Limsampan and Patcharapan Suwannin and Mongkol Techakasikornpanich and Galayanee Doungchawee and Edilberto Manahan and Kulachart Jangpatarapongsa},
journal = {Journal of clinical laboratory analysis},
year = {2026},
month = oct,
doi = {10.1002/jcla.70277},
url = {https://europepmc.org/article/MED/42876165},
note = {CC BY},
}
RIS
TY - JOUR
TI - Novel Target for Pathogenic Leptospira Detection via Colorimetric Loop-Mediated Isothermal Amplification (LAMP): Development, Validation, and Comparative Analysis.
AU - Peyman Ghoraishizadeh
AU - Benjie Clemente
AU - Kween Saimuang
AU - Sakaorat Lertjuthaporn
AU - Sukhonta Limsampan
AU - Patcharapan Suwannin
AU - Mongkol Techakasikornpanich
AU - Galayanee Doungchawee
AU - Edilberto Manahan
AU - Kulachart Jangpatarapongsa
PY - 2026
DA - 2026/10/14
JO - Journal of clinical laboratory analysis
DO - 10.1002/jcla.70277
UR - https://europepmc.org/article/MED/42876165
AB - Background Leptospirosis is a globally prevalent zoonotic disease that remains challenging to diagnose due to its nonspecific symptoms and the limitations of current diagnostic methods and targets. To address these challenges, we developed and validated a naked-eye loop-mediated isothermal amplification (LAMP) assay targeting the newly identified orpH region for the rapid detection of pathogenic Leptospira species.Methods Comparative genomic analysis identified orpH as a conserved sequence among pathogenic Leptospira. A colorimetric LAMP assay (LAMP-orpH) was developed and evaluated using reference strains and serum samples obtained from patients with suspected leptospirosis. The diagnostic performance of the assay on these serum samples was evaluated in comparison with conventional PCR and the microscopic agglutination test (MAT), with the real-time PCR assay targeting lipL32 designated as the reference standard.Results The LAMP-orpH assay demonstrated 87.2% sensitivity and 94.8% specificity among 76 suspected cases and 29 controls, showing strong agreement with real-time PCR (κ = 0.826). It outperformed MAT (63.8% sensitivity) and conventional PCR (23.4% sensitivity), achieving a detection limit of 10² cells/mL in spiked serum.Conclusion The LAMP-orpH assay offers a rapid, sensitive, and specific tool for early leptospirosis diagnosis. Its simple colorimetric readout and minimal equipment requirements make it ideal for use in rural and resource-limited settings, providing a practical alternative to MAT, conventional PCR, and qPCR.
SN - 0887-8013
SN - 1098-2825
ER -