Development of Quick Real-time PCR Diagnostic Method for Canine Infectious Dermatitis using GeneChecker

Abstract

Canine dermatitis is one of the most important diseases, affecting about 10% of dogs. Infectious dermatitis in dogs is caused by a primary or secondary infection with bacteria, fungi, including yeast, and ectoparasites. Diagnosis of this infectious dermatitis consists of physical examination, clinical symptoms, microscopic examination of samples from the lesion site, culture and identification examination, and molecular biological diagnosis. This study was performed to develop a quick real-time PCR kit using a chip for the four main pathogens of infectious dermatitis (Staphylococcus pseudintermedius, Microsporum canis, Malassezia sp., and Demodex canis). Skin and hair samples from canine dermatitis patients who visited the Veterinary Medical Teaching Hospital at Jeju National University were collected and pathogens were identified through microscopic examination, Diff-Quik staining, and culture isolation. As a result of comparing conventional PCR and quick real-time PCR tests using nuc, dp5, nad4 and chs gene-specific primers for S. pseudintermedius, M. canis, Malassezia sp. and D. canis, respectively. It was possible to detect the causative pathogen being detected without cross-reacting with other pathogens. In particular, the quick real-time PCR test was more convenient and quicker than the conventional PCR. In this study, through the development of the chip-based quick real-time PCR diagnostic method, it is thought that will be a useful point-of-care test(POCT) that can be applied to easily and quickly diagnose the causative bacteria of skin diseases in general veterinary hospitals, not laboratories, and to check the response to treatment.