Background: Rapid and accurate laboratory diagnoses of viral infections are crucial for the management and treatment of patients with viral infections. Conventional methods for virus detection are labourious, time consuming, and only a single virus can be analysed in one assay. Objectives: The objective of this study was to develop a novel real-time PCR method for multiple virus detection by melting curve analysis using Taqman probes in a single reaction. Study design: As a model, six respiratory viruses were detected in one tube using three fluorophores. The specificity was assessed by cross-reaction tests with other common respiratory pathogens. The analytical sensitivity was assessed by testing the limit of detection of the assay using artificial plasmids as the positive template. The clinical evaluation of the established assay was evaluated for the detection of respiratory viruses in clinical samples, and the results were compared with direct fluorescent antibody testing (DFA). Results: The six respiratory viruses were clearly distinguished by their respective melting temperature values in the corresponding fluorescence detection channels. No cross reactions were observed by cross reaction tests. The detection limits of this assay were 2 to 2 x 10(3) copies per reaction for each virus. The clinical evaluation of this assay was demonstrated by analysing 352 clinical samples, and 67(19.0%) samples were positive for at least one virus. The accordance rate between the established PCR and DFA testing was high, and ranged from 94.57% to 100%. Conclusions: Taqman probe-based melting curve analysis is well suited for detection of multiple viruses in clinical and research laboratories because of its high throughput, reliability, and cost savings.
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[1]
Botezatu I V, 2017, Mol Biol (Mosk), V51, P50, DOI 10.7868/S0026898417010062
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Boyd, Victoria
Smith, Ina
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CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Smith, Ina
Crameri, Gary
论文数: 0引用数: 0
h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Crameri, Gary
Burroughs, Amy L.
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Burroughs, Amy L.
Durr, Peter A.
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Durr, Peter A.
White, John
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
White, John
Cowled, Christopher
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Cowled, Christopher
Marsh, Glenn A.
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Marsh, Glenn A.
Wang, Lin-Fa
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, Australia
Duke NUS Grad Med Sch Singapore, Program Emerging Infect Dis, Singapore, SingaporeCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Kishimoto, Mai
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Tsuchiaka, Shinobu
Rahpaya, Sayed Samim
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h-index: 0
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Rahpaya, Sayed Samim
Hasebe, Ayako
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h-index: 0
机构:
Gifu Cent Livestock Hyg Serv Ctr, 4-2-22 Imarnine, Gifu 5008388, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Hasebe, Ayako
Otsu, Keiko
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h-index: 0
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Gifu Cent Livestock Hyg Serv Ctr, 4-2-22 Imarnine, Gifu 5008388, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Otsu, Keiko
Sugimura, Satoshi
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h-index: 0
机构:
Tokyo Univ Agr & Technol, Inst Agr, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Sugimura, Satoshi
Kobayashi, Suguru
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Tokyo Univ Agr & Technol, Inst Agr, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Kobayashi, Suguru
Komatsu, Natsumi
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h-index: 0
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Tokyo Univ Agr & Technol, Inst Agr, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Komatsu, Natsumi
Nagai, Makoto
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h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Ishikawa Prefectural Univ, Fac Bioresources & Environm Sci, 1-308 Suematsu, Nonoichi, Ishikawa 9218836, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Nagai, Makoto
Omatsu, Tsutomu
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Omatsu, Tsutomu
Naoi, Yuki
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Naoi, Yuki
Sano, Kaori
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Sano, Kaori
Okazaki-Terashima, Sachiko
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Okazaki-Terashima, Sachiko
Oba, Mami
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Oba, Mami
Katayama, Yukie
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Katayama, Yukie
Sato, Reiichiro
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h-index: 0
机构:
Azabu Univ, Sch Vet Med, Chuo Ku, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 2525201, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Sato, Reiichiro
Asai, Tetsuo
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Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Asai, Tetsuo
Mizutani, Tetsuya
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Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Boyd, Victoria
Smith, Ina
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Smith, Ina
Crameri, Gary
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Crameri, Gary
Burroughs, Amy L.
论文数: 0引用数: 0
h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Burroughs, Amy L.
Durr, Peter A.
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Durr, Peter A.
White, John
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
White, John
Cowled, Christopher
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h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Cowled, Christopher
Marsh, Glenn A.
论文数: 0引用数: 0
h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, AustraliaCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Marsh, Glenn A.
Wang, Lin-Fa
论文数: 0引用数: 0
h-index: 0
机构:
CSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
Australian Anim Hlth Lab, Geelong, Vic 3220, Australia
Duke NUS Grad Med Sch Singapore, Program Emerging Infect Dis, Singapore, SingaporeCSIRO Biosecur Flagship, Geelong, Vic 3220, Australia
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Kishimoto, Mai
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h-index:
机构:
Tsuchiaka, Shinobu
Rahpaya, Sayed Samim
论文数: 0引用数: 0
h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Rahpaya, Sayed Samim
Hasebe, Ayako
论文数: 0引用数: 0
h-index: 0
机构:
Gifu Cent Livestock Hyg Serv Ctr, 4-2-22 Imarnine, Gifu 5008388, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Hasebe, Ayako
Otsu, Keiko
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h-index: 0
机构:
Gifu Cent Livestock Hyg Serv Ctr, 4-2-22 Imarnine, Gifu 5008388, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Otsu, Keiko
Sugimura, Satoshi
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h-index: 0
机构:
Tokyo Univ Agr & Technol, Inst Agr, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Sugimura, Satoshi
Kobayashi, Suguru
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h-index: 0
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Tokyo Univ Agr & Technol, Inst Agr, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Kobayashi, Suguru
Komatsu, Natsumi
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h-index: 0
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Tokyo Univ Agr & Technol, Inst Agr, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Komatsu, Natsumi
Nagai, Makoto
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h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Ishikawa Prefectural Univ, Fac Bioresources & Environm Sci, 1-308 Suematsu, Nonoichi, Ishikawa 9218836, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Nagai, Makoto
Omatsu, Tsutomu
论文数: 0引用数: 0
h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Omatsu, Tsutomu
Naoi, Yuki
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h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Naoi, Yuki
Sano, Kaori
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h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Sano, Kaori
Okazaki-Terashima, Sachiko
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h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Okazaki-Terashima, Sachiko
Oba, Mami
论文数: 0引用数: 0
h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Oba, Mami
Katayama, Yukie
论文数: 0引用数: 0
h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Katayama, Yukie
Sato, Reiichiro
论文数: 0引用数: 0
h-index: 0
机构:
Azabu Univ, Sch Vet Med, Chuo Ku, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 2525201, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Sato, Reiichiro
Asai, Tetsuo
论文数: 0引用数: 0
h-index: 0
机构:
Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Asai, Tetsuo
Mizutani, Tetsuya
论文数: 0引用数: 0
h-index: 0
机构:
Tokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan
Gifu Univ, United Grad Sch Vet Sci, 1-1 Yanagito, Gifu 5011193, JapanTokyo Univ Agr & Technol, Res & Educ Ctr Prevent Global Infect Dis Anim, 3-5-8 Saiwai, Fuchu, Tokyo 1838509, Japan