Journal of Veterinary Science & Medical Diagnosis ISSN: 2325-9590

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Research Article, J Vet Sci Med Diagn Vol: 4 Issue: 2

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

Recavarren M1-3*, Quintana S2, Rivero M3, Ballesteros B3 and Scialfa E4
1Veterinary Areas of Fares Taie Biochemical Analysis Institute, Argentina
2Molecular Biology Areas of Fares Taie Biochemical Analysis Institute, Argentina
3Faculty of Veterinary Science, UNCPBA, Tandil, Argentina
4División Zoonosis Rurales de Azul. Rivadavia 3331, (7600) Mar del Plata, 54-223-4753855 (Int.133), Argentina
Corresponding author : Mariana Recavarren
Rivadavia 3331, (7600) Mar del Plata, Argentina
Tel: 54-223-4753855 (Int.133)
E-mail: [email protected]
Received: July 11, 2014 Accepted: November 10, 2014 Published: November 14, 2014
Citation: Recavarren M, Quintana S, Rivero M, Ballesteros B, Scialfa E (2015) Leptospira Diagnosis in Serum and Urine of Dogs by Real Time PCR. J Vet Sci Med Diagn 4:2. doi:10.4172/2325-9590.1000157

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.

Keywords: Leptospirosis; Canine; L-MAT; Real time PCR

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