Immunology and Infectious Diseases(CEASE PUBLICATION) Vol. 4(4), pp. 39 - 43
DOI: 10.13189/iid.2016.040401
Reprint (PDF) (514Kb)


Detection of Pathogenic Leptospiral DNA Traces in Canine Sera Serum Samples by Loop Mediated Isothermal Amplification (LAMP)


Sylvia Grune Loffler 1,2,*, Carlos Leiva 1,2, Exequiel Scialfa 3, Leandro Redondo 1,2, Mónica Florin-Christensen 1,2, Mara Martínez 1, Graciela Romero 1, Bibiana Brihuega 1
1 Institute of Pathobiology, Laboratory of Leptospirosis, National Institute of Agricultural Technology, Argentina
2 National Research Council of Argentina (CONICET), Buenos Aires, Argentina
3 Division of Rural Zoonosis, Azul, Buenos Aires province, Argentina

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.

KEYWORDS
Leptospirosis, Serum, LAMP, Diagnostics, Canine

Cite This Paper in IEEE or APA Citation Styles
(a). IEEE Format:
[1] Sylvia Grune Loffler , Carlos Leiva , Exequiel Scialfa , Leandro Redondo , Mónica Florin-Christensen , Mara Martínez , Graciela Romero , Bibiana Brihuega , "Detection of Pathogenic Leptospiral DNA Traces in Canine Sera Serum Samples by Loop Mediated Isothermal Amplification (LAMP)," Immunology and Infectious Diseases(CEASE PUBLICATION), Vol. 4, No. 4, pp. 39 - 43, 2016. DOI: 10.13189/iid.2016.040401.

(b). APA Format:
Sylvia Grune Loffler , Carlos Leiva , Exequiel Scialfa , Leandro Redondo , Mónica Florin-Christensen , Mara Martínez , Graciela Romero , Bibiana Brihuega (2016). Detection of Pathogenic Leptospiral DNA Traces in Canine Sera Serum Samples by Loop Mediated Isothermal Amplification (LAMP). Immunology and Infectious Diseases(CEASE PUBLICATION), 4(4), 39 - 43. DOI: 10.13189/iid.2016.040401.