Differential Detection of Hemotropic Mycoplasma Species in Cattle by Melting Curve Analysis of the PCR Products
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概要
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A real-time PCR procedure followed by melting curve analysis, using the green fluorescence dye SYBR Green I, was developed for rapid detection and differentiation of hemplasmas in cattle. Analysis of the melting temperature (Tm) of the PCR products allowed to differentiation of the two bovine hemoplasmas, Mycoplasma wenyonii and a provisional species, ‘Candidatus Mycoplasma haemobovis’ (synonym of ‘Candidatus M. haemobos’). The Tm (mean±SE) of the PCR products from bovine hemoplasmas were 86.98±0.12℃ for M. wenyonii, and 82.04±0.27℃ for ‘Candidatus M. haemobovis’ in the melting experiments. The protocol described in the present study can decrease the time to results by simultaneous detection and differentiation of the two hemoplasmas in cattle. By using this protocol, we examined hemoplasma prevalence in 109 cattle in the Miyagi prefecture, and found that 67 (61.5%) were infected with M. wenyonii, 25 (22.9%) were with ‘Candidatus M. haemobovis’, and 14 (12.8%) were mixed infection.
著者
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SATO Makoto
Department of Applid Chemistry, Graduate School of Engineering, Tohoku University
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FUJIHARA Masatoshi
Department of Veterinary Microbiology, School of Veterinary Medicine, Faculty of Agriculture, Iwate
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Sato Shigeru
Department Of Gastroenterology Fukuoka University Chikushi Hospital
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Harasawa Ryo
Department Of Veterinary Medicine Faculty Of Agriculture Iwate University
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NISHIZAWA Ikuo
Department of Veterinary Microbiology, School of Veterinary Medicine, Faculty of Agriculture, Iwate
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Sato Makoto
Department Of Applid Chemistry Graduate School Of Engineering Tohoku University
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FUJIHARA Masatoshi
Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, Morioka 020–8550, Japan
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NISHIZAWA Ikuo
Department of Veterinary Microbiology, Faculty of Agriculture, Iwate University
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NISHIZAWA Ikuo
Department of Veterinary Microbiology, School of Veterinary Medicine, Faculty of Agriculture, Iwate University
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Sato Shigeru
Department of Chemistry, Faculty of Science, Tokyo Institute of Technology
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SATO Shigeru
Department of Clinical Veterinary Medicine, School of Veterinary Medicine, Faculty of Agriculture, Iwate University
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