- 著者
- Mai KISHIMOTO Shinobu TSUCHIAKA Sayed Samim RAHPAYA Ayako HASEBE Keiko OTSU Satoshi SUGIMURA Suguru KOBAYASHI Natsumi KOMATSU Makoto NAGAI Tsutomu OMATSU Yuki NAOI Kaori SANO Sachiko OKAZAKI-TERASHIMA Mami OBA Yukie KATAYAMA Reiichiro SATO Tetsuo ASAI Tetsuya MIZUTANI
- 出版者
- 公益社団法人 日本獣医学会
- 雑誌
- Journal of Veterinary Medical Science (ISSN:09167250)
- 巻号頁・発行日
- pp.16-0489, (Released:2017-01-08)
- 被引用文献数
- 56
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.
- 著者
- Karla Cristine C. DOYSABAS Mami OBA Tomoki ISHIBASHI Hideki SHIBATA Hitoshi TAKEMAE Hiroshi SHIMODA Tetsuya MIZUTANI Atsuo IIDA Eiichi HONDO
- 出版者
- JAPANESE SOCIETY OF VETERINARY SCIENCE
- 雑誌
- Journal of Veterinary Medical Science (ISSN:09167250)
- 巻号頁・発行日
- pp.20-0021, (Released:2020-02-12)
- 被引用文献数
- 2
Adenosine 5’-triphosphate (ATP), the major energy currency of the cell, is involved in many cellular processes, including the viral life cycle, and can be used as an indicator of early signs of cytopathic effect (CPE). In this study, we demonstrated that CPE can be analyzed using an FRET-based ATP probe named ATP indicator based on Epsilon subunit for Analytical Measurements (ATeam). The results revealed that as early as 3 hr, the virus infected cells showed a significantly different Venus/cyan fluorescent protein (CFP) ratio compared to the mock-infected cells. The ATeam technology is therefore useful to determine the early signs of ATP-based CPE as early as 3 hr without morphology-based CPE by light microscopy, and enables high throughput determination of the presence of microorganisms in neglected samples stored in laboratories.
- 著者
- Mami OBA Tsutomu OMATSU Ai TAKANO Hiromi FUJITA Kozue SATO Atsushi NAKAMOTO Mamoru TAKAHASHI Nobuhiro TAKADA Hiroki KAWABATA Shuji ANDO Tetsuya MIZUTANI
- 出版者
- 公益社団法人 日本獣医学会
- 雑誌
- Journal of Veterinary Medical Science (ISSN:09167250)
- 巻号頁・発行日
- pp.15-0536, (Released:2015-10-25)
- 被引用文献数
- 1 25
Soft ticks, Argas vespertilionis, were collected from feces of bats in Japan. Cytopathic effect (CPE) was observed after inoculating the homogenates of ticks to Vero cells. Sequencing of RNA extracted from the cell supernatant was performed by next generation sequencer. The contigs had identity to segments of Bunyaviruses, Issyk-Kul virus. The identities of segment L, M and S were only 77, 76 and 79% to Issyk-Kul virus, respectively. Therefore, we named this novel virus Soft tick bunyavirus (STBV). In the phylogenetic tree, segment L of STBV was closely related to a cluster consisting of the genus Nairovirus of the family Bunyaviridae.