Diagnosis of Canine Leptospirosis by a Highly Sensitive FRET-PCR Targeting the lig Genes

摘要: Canine leptospirosis is underdiagnosed due to its wide spectrum of clinical presentations and the lack a rapid sensitive test for accurate diagnosis acute chronic infections. In this study, we developed highly specific fluorescence resonance energy transfer (FRET)-PCR detect common pathogenic leptospires in dogs, including Leptospira interrogans serovars Autumnalis, Canicola, Copenhageni (Icterohaemorrhagiae serogroup) Pomona, kirschneri serovar Grippotyphosa. This PCR targets lig genes, exclusively found species but not saprophytic (L. biflexa). A robust, high-stringency step-down real-time platform was coupled detection leptospiral DNA by fluorescently labeled FRET probes. enabled single copy gene containing from up 50 µL canine blood or 400 urine. Sensitivity determination use limiting serial dilutions extracted indicated that FRET-PCR established almost 100-fold more than widely accepted lipL32 SYBR assay 10-fold 16S rRNA TaqMan assay. Application method 207 dogs with potential infection us diagnose three cases characterized low amounts body fluids. Detection PCR, which detected only 2 3 cases, none leptospirosis.