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KMID : 1023520230460010015
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Korean Journal of Veterinary Service
2023 Volume.46 No. 1 p.15 ~ p.27
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A triplex real-time PCR assay for simultaneous and differential detection of Bordetella bronchiseptica, Mycoplasma cynos, and Mycoplasma canis in respiratory diseased dogs
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Jeon Gyu-Tae
Kim Jong-Min
Park Jeong-Hyun
Kim Hye-Ryung
Baek Ji-Su
Lee Hyo-Ji
Shin Yeun-Kyung
Kwon Oh-Kyu
Kang Hae-Eun
Kim Soong-Koo
Kim Jung-Hwa
Kim Young-Hwan
Park Choi-Kyu
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Abstract
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Bordetella (B.) bronchiseptica, Mycoplasma (M.) cynos, and M. canis are the major bacterial pathogens that cause canine infectious respiratory disease complex (CIRDC). In this study, we developed a triplex real-time polymerase chain reaction (tqPCR) assay for the differential detection of these bacteria in a single reaction. The assay specifically amplified three bacterial genes with a detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra- and inter-assay variations of less than 1%. The diagnostic results of the assay using 94 clinical samples from household dogs with CIRDC clinical signs, the prevalence of B. bronchiseptica, M. cynos, and M. canis was 22.3%, 18.1%, and 20.2%, respectively, indicating that the diagnostic sensitivity was comparable to those of previously reported qPCR assays. The dual infection rate of B. bronchiseptica and M. cynos, B. bronchiseptica and M. canis, and M. cynos and M. canis was 5.3%, 7.4%, and 3.1%, respectively. Moreover, the triple infection rate of B. bronchiseptica, M. cynos, and M. canis was 2.1%. These results indicate that coinfections with B. bronchiseptica, M. cynos, and M. canis have frequently occurred in the Korean dog population. The newly developed tqPCR assay in the present study will be a useful tool for etiological and epidemiological studies on these three CIRDC-associated bacterial pathogens. The prevalence and coinfection data revealed through this study will contribute to expanding knowledge on the epidemiology of CIRDC in the recent Korean dog population.
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KEYWORD
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Triplex real-time PCR, Dogs, Bordetella bronchiseptica, Mycoplasma cynos, Mycoplasma canis
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