01352-A: Detection of Brucella canis DNA in canine urine, semen and vaginal cells via qPCR Analysis

Grant Status: Closed

Grant Amount: $12,960
Dr. Lin Kauffman, D.V.M., Iowa State University
August 1, 2009 - July 31, 2010
Sponsor(s): Akita Club of America, Inc., American Brittany Club, American Cavalier King Charles Spaniel Club Charitable Trust, American Chesapeake Bay Retriever Club, American German Shepherd Dog Charitable Foundation, Inc., American Shih Tzu Club, Inc., Basenji Club of America, Inc. & Basenji Health Endowment, Border Terrier Club of America, Cairn Terrier Club of America, English Springer Spaniel Field Trial Association, German Wirehaired Pointer Club of America, National Beagle Club, Papillon Club of America, Scottish Terrier Club of America, Small Munsterlander Club of North America, Standard Schnauzer Club of America, Vizsla Club of America Welfare Foundation, Yorkshire Terrier Club of America, Yorkshire Terrier Club of America Foundation, Inc.
Breed(s): -All Dogs
Research Program Area: Treatment

Project Summary

Brucella canis, a bacteria that causes reproductive disorders in dogs, has been on the increase across the US. This goes hand in hand with increasing costs associated with reproduction losses due to disease and euthanasia of infected animals. The potential for human infection from dogs increases concern over this infectious disease. Currently the only available tests for this disease are not great. The bacteria is hard to culture and serological tests detect the disease 8-12 weeks post-infection but cannot detect early disease. Present laws make canine brucellosis a reportable disease subject to quarantine. In some states this means testing all breeding animals and euthanasia of infected animals. Early detection of disease would shorten the quarantine period and would add a measure of safely for buyers of puppies or breeding stock. The goal of this project is to use Polymerase Chain Reaction (qPCR) assay to detect Brucella sp. in samples of suspect B. canis urine (male), semen and vaginal cells (female) and compare how this test works vs. traditional culture and serology. To find a more specific and timely diagnostic, this study assessed the ability of qPCR analysis to detect B. canis Omp25 DNA in a variety of samples (blood, urine, vaginal swab) and compared those results against current detection methods for B. canis infection in dogs (serology). qPCR analysis identified the presence of B.canis Omp25 DNA in multiple dogs prior to seroconversion. Non-invasive samples from the genito-urinary tract, including vaginal swabs and urine, were found to be the most sensitive for detection of B. canis Omp25 DNA via qPCR. Use of these samples would make collection of diagnostic samples within the ability of some dog owners and breeders. The results of this study are very encouraging for use of B. canis Omp25-specific qPCR as a diagnostic screening tool for B. canis. The potential of this assay qPCR for early detection could be very valuable for elimination of B. canis from kennels without having to wait for seroconversion. Additionally, Omp25 qPCR could be a valuable screening tool for B. canis in newly purchased dogs prior to adding these dogs into a new home or kennel. B. canis is a reemerging infectious disease in the canine breeding industry. A better screening and detection method will be very useful to prevent further spread of this insidious disease. B. canis Omp25 qPCR may be this critical diagnostic component to decrease economic effects of canine brucellosis on the canine breeding industry and prevalence of canine brucellosis in the US.

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