Development of a one-run real-time PCR detection system for pathogens associated with bovine respiratory disease complex

DOI Web Site PubMed 被引用文献10件 参考文献29件
  • KISHIMOTO Mai
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan
  • TSUCHIAKA Shinobu
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagito, Gifu 501-1193, Japan
  • RAHPAYA Sayed Samim
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagito, Gifu 501-1193, Japan
  • HASEBE Ayako
    Gifu Central Livestock Hygiene Service Center, 4-2-22 Imamine, Gifu, Gifu 500-8388, Japan
  • OTSU Keiko
    Gifu Central Livestock Hygiene Service Center, 4-2-22 Imamine, Gifu, Gifu 500-8388, Japan
  • SUGIMURA Satoshi
    Institute of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183-8509, Japan
  • KOBAYASHI Suguru
    Institute of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183-8509, Japan
  • KOMATSU Natsumi
    Institute of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183-8509, Japan
  • NAGAI Makoto
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan Faculty of Bioresources and Environmental Sciences, Ishikawa prefectural University, 1-308, Suematsu, Nonoichi-shi, Ishikawa 921-8836, Japan
  • OMATSU Tsutomu
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagito, Gifu 501-1193, Japan
  • NAOI Yuki
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan
  • SANO Kaori
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan
  • OKAZAKI-TERASHIMA Sachiko
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan
  • OBA Mami
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan
  • KATAYAMA Yukie
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan
  • SATO Reiichiro
    Azabu University, School of Veterinary Medicine, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan
  • ASAI Tetsuo
    The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagito, Gifu 501-1193, Japan
  • MIZUTANI Tetsuya
    Research and Education Center for Prevention of Global Infectious Diseases of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183–8509, Japan The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagito, Gifu 501-1193, Japan

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<p>Bovine respiratory disease complex (BRDC) is frequently found in cattle worldwide. The etiology of BRDC is complicated by infections with multiple pathogens, making identification of the causal pathogen difficult. Here, we developed a detection system by applying TaqMan real-time PCR (Dembo respiratory-PCR) to screen a broad range of microbes associated with BRDC in a single run. We selected 16 bovine respiratory pathogens (bovine viral diarrhea virus, bovine coronavirus, bovine parainfluenza virus 3, bovine respiratory syncytial virus, influenza D virus, bovine rhinitis A virus, bovine rhinitis B virus, bovine herpesvirus 1, bovine adenovirus 3, bovine adenovirus 7, Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, Trueperella pyogenes, Mycoplasma bovis and Ureaplasma diversum) as detection targets and designed novel specific primer-probe sets for nine of them. The assay performance was assessed using standard curves from synthesized DNA. In addition, the sensitivity of the assay was evaluated by spiking solutions extracted from nasal swabs that were negative by Dembo respiratory-PCR for nucleic acids of pathogens or synthesized DNA. All primer-probe sets showed high sensitivity. In this study, a total of 40 nasal swab samples from cattle on six farms were tested by Dembo respiratory-PCR. Dembo respiratory-PCR can be applied as a screening system with wide detection targets.</p>

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