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Evaluation of loop-mediated isothermal amplification method (LAMP) for pathogenic Leptospira spp. detection with
Duangjai Suwancharoen1, Busara Sittiwicheanwong, Anuwat Wiratsudakul
1National Institute of Animal Health, Department of Livestock Development, 50/2 Kasetklang, Ladyao, Chatuchak, Bangkok 10900, Thailand.
Abstract:
Leptospirosis has been one of the worldwide zoonotic diseases caused by pathogenic Leptospira spp. Many molecular techniques have consecutively been developed to detect such pathogen including loop-mediated isothermal amplification method (LAMP). The objectives of this study were to evaluate the diagnostic accuracy of LAMP assay and real-time PCR using bacterial culture as the gold standard and to assess the agreement among these three tests using Cohen's kappa statistics. In total, 533 urine samples were collected from 266 beef and 267 dairy cattle reared in central region of Thailand. Sensitivity and specificity of LAMP were 96.8% (95% CI 81.5-99.8) and 97.0% (95% CI 94.9-98.2), respectively. The accuracy of LAMP (97.0%) was significantly higher than that of real-time PCR (91.9%) at 95% CI. With Cohen's kappa statistics, culture method and LAMP were substantially agreed with each other (77.4%), whereas real-time PCR only moderately agreed with culture (47.7%) and LAMP (45.3%), respectively. Consequently, LAMP was more effective than real-time PCR in detecting Leptospira spp. in the urine of cattle. Besides, LAMP had less cost and was simpler than real-time PCR. Thus, LAMP was an excellent alternative for routine surveillance of leptospirosis in cattle.
Insights
Loop-mediated isothermal amplification (LAMP) is a highly accurate and cost-effective method for detecting Leptospira spp. in cattle, outperforming real-time PCR in surveillance.
Area of Science:
- Veterinary Microbiology
- Zoonotic Diseases
- Molecular Diagnostics
Background:
- Leptospirosis, a global zoonotic disease, is caused by pathogenic Leptospira species.
- Accurate detection of Leptospira is crucial for controlling disease spread in cattle populations.
Purpose of the Study:
- To evaluate the diagnostic accuracy of the loop-mediated isothermal amplification (LAMP) assay compared to real-time PCR and bacterial culture for Leptospira detection in cattle.
- To assess the agreement between LAMP, real-time PCR, and bacterial culture using Cohen's kappa statistics.
Main Methods:
- Analysis of 533 cattle urine samples from central Thailand using bacterial culture (gold standard), LAMP, and real-time PCR.
- Calculation of sensitivity, specificity, and accuracy for LAMP and real-time PCR.
- Assessment of inter-test agreement using Cohen's kappa statistics.
Main Results:
- LAMP demonstrated high sensitivity (96.8%) and specificity (97.0%).
- LAMP's overall accuracy (97.0%) was significantly higher than real-time PCR (91.9%).
- Substantial agreement was found between LAMP and bacterial culture (kappa=0.774), while moderate agreement was observed for real-time PCR with culture (kappa=0.477) and LAMP (kappa=0.453).
Conclusions:
- LAMP is a more effective diagnostic tool than real-time PCR for detecting Leptospira in cattle urine.
- LAMP offers a simpler, less expensive, and excellent alternative for routine leptospirosis surveillance in cattle.
- The findings support the widespread adoption of LAMP for improved cattle health management.
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