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1 Bulletin of the Veterinary Institute in Puławy

1 2000

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Application of PCR and nested-PCR for detection of Borrelia burgdorferi in experimentally infected dogs

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Mizak B. , Krol J. , Rzezutka A.

Bulletin of the Veterinary Institute in Puławy

2000

tom 44

nr 1

The aim of these studies was to attempt to infect dogs by their inoculation with cultured Borrelia burgdorferi isolated from ticks in Poland and to use PCR and N-PCR to evaluate the spread of infection. Polish strain 236/2 of B. burgdorferi isolated from Ixodes ricinus and cultured in BSK-H medium (passage 4 and 5) was used for dog inoculation. Throughout the experimental trial all dogs, including the control, remained healthy. None of them became hyperthermic nor demonstrated clinical signs of the disease, but seroconversion was observed. During our experiments it was demonstrated that the culture of spirochete in BSK-H medium is time consuming, does not always yield spirochetes and requires up to 4 weeks growth time. Application of PCR and N-PCR confirmed the replication of B. burgdorferi in infected dogs. Spirochetes were demonstrated almost in all internal organs of challenged animals as well as in skin, muscles, adipose tissue, joints and urine. Results of our studies indicated, that PCR and N-PCR can be used for detection of B. burgdorferi both in infected BSK-H medium or internal organs of challenged dogs. The high sensitivity and specificity of both tests enable tchem to be used for laboratory diagnosis of dog borreliosis and for monitoring of the spread of infection within the animal.