Abstract
Leptospirosis is a worldwide distributed zoonosis, caused by pathogenic strains of spirochetes belonging to the genus Leptospira spp. So far, the only gold standard diagnostic method for this disease has been the Micro-Agglutination Test (MAT). The purpose of this study was to test whether a correlation exists between serological diagnosis by MAT and the presence of Leptospira spp. DNA traces in serum. Anti-Leptospira spp. antibody titers were determined in dog serum samples obtained at a canine shelter from Buenos Aires Province, Argentina (n= 80) by MAT using ten Leptospira spp. serovars. DNA extracted from these samples using Chelex-100 resin was used as template for Leptospira spp. specific PCR and loop-mediated isothermal amplification (LAMP) tests. In the latter case, product detection by incubation with malachite green dye was set up, obtaining optimal results at a dye concentration of 0.02% (w(v)) and a pH of 6.1. PCR and LAMP showed the same sensitivity, detecting leptospiral DNA in 12 out of the 48 MAT- positive sera. No detection was attained by either method in MAT-negative samples. These results show that it is possible to use LAMP for the detection of leptospirosis in non-human mammals. It also shows that, in the case of serum samples, MAT has higher diagnostic sensitivity than the currently available PCR and LAMP tests.
Highlights
Pathogenic strains of Leptospira spp. belong to the order Spirochaetales, family Leptospiraceae and are the causative bacterial agent of leptospirosis
Domestic and wild animals can be infected with pathogenic Leptospira spp
Pathogenic Leptospira spp. strains isolated so far from dogs in Argentina have been serotyped by Cross Agglutination Absorption test (CAAT) belong to the L. interrogans species and the Canicola, Icterohaemorrhagiae and Pyrogenes serogroups [8]
Summary
Pathogenic strains of Leptospira spp. belong to the order Spirochaetales, family Leptospiraceae and are the causative bacterial agent of leptospirosis This disease is the most widespread zoonosis in the world, being endemic in most tropical and subtropical regions. Pathogenic Leptospira spp. strains isolated so far from dogs in Argentina have been serotyped by Cross Agglutination Absorption test (CAAT) belong to the L. interrogans species and the Canicola, Icterohaemorrhagiae and Pyrogenes serogroups [8]. LAMP was applied to the detection of Anaplasma phagocytophilum, Babesia canis canis, Echinococcus granulosus, canine parvovirus and canine distemper virus [17] This technology has been tested for the detection of Leptospira spp. in human urine and blood samples [18, 19, 20, 21], but no studies have so far used LAMP to detect pathogenic Leptospira spp. in animal samples. We tested whether it was possible to detect traces of pathogenic leptospiral DNA in canine serum samples by PCR and LAMP, and analyzed the diagnostic potential of these methods as compared to MAT
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