Evaluation of loop-mediated isothermal amplification method (LAMP) for pathogenic Leptospira spp. detection with leptospires isolation and real-time PCR.

Publisher: Japanese Society Of Veterinary Science Country of Publication: Japan NLM ID: 9105360 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1347-7439 (Electronic) Linking ISSN: 09167250 NLM ISO Abbreviation: J Vet Med Sci Subsets: MEDLINE

Original Publication: Tokyo : Japanese Society Of Veterinary Science

Leptospira*/genetics; Cattle Diseases/*diagnosis ; Leptospirosis/*veterinary ; Nucleic Acid Amplification Techniques/*veterinary ; Real-Time Polymerase Chain Reaction/*veterinary; Animals ; Cattle ; Cattle Diseases/microbiology ; Leptospirosis/diagnosis ; Leptospirosis/microbiology ; Nucleic Acid Amplification Techniques/methods ; Real-Time Polymerase Chain Reaction/methods ; Reproducibility of Results ; Sensitivity and Specificity

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.

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Date Created: 20160507 Date Completed: 20170615 Latest Revision: 20181113

20221213

PMC5053931

10.1292/jvms.15-0702

27150208