Leptospira Diagnosis in Serum and Urine of Dogs by Real Time PCR

Abstract

Leptospira Diagnosis in Serum and Urine of Dogs by Real Time PCR A real-time PCR assay to detect DNA of Leptospira interrogans serovars infecting dogs from pure cultures and clinical samples was developed. DNA amplification from pure cultures of serogroups Canicola, Pomona, Pyrogenes, Ballum and Icterohaemorrhagiae was performed by real-time PCR showing an analytical sensitivity of 8 genomes. From 70 sera samples of suspected canine leptospirosis, microscopic agglutination test (MAT) was performed to determine antibodies and realtime PCR to detect leptospiral DNA, dog DNA internal control amplification was performed in all samples to verify successful DNA extraction. Thirteen samples were positive by real-time PCR and L-MAT ≤ 1/200 and ≥ 1/400 with co-agglutination which represents 7 and 10 days post infections. One urine sample was positive by real-time PCR which serum was prior negative to MAT. The real-time PCR assay could successfully detect leptospiral DNA in clinical samples from dogs in various stages of infection and can be used to complement MAT technique.