Development of serological assays for determining MERS coronavirus neutralizing antibody responses

• Project/Area Number: 26450418
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: FUKUSHI SHUETSU 国立感染症研究所, ウイルス第一部, 主任研究官 (80373398)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: MERSコロナウイルス / 血清疫学 / モノクローナル抗体 / 競合ELISA / MERS-CoV / 血清疫学研究 / 抗体検出 / VSVシュードタイプ / 抗体測定

Outline of Final Research Achievements

Since discovering the MERS coronavirus (MERS-CoV) as a causative agent of severe respiratory illness in the middle east in 2012, a serological antibody testing has been conducted not only for assessing antibody responses of infected patients but also for investigating zoonotic origins of MERS-CoV. In this study, we developed a high-throughput virus neutralization assay using pseudotyped vesicular stomatitis virus (VSV)-bearing MERS-CoV S protein (MERSpv). In addition, we generated monoclonal antibody（MAb）against S protein, and developed an MAb-based competitive ELISA (cELISA) to detect MERS-CoV antibody. Both VSV pseudotype-based and cELISA are useful for the epidemiological investigation on MERS-CoV infection.

Research Products

• [Journal Article] Characterization of novel monoclonal antibodies against the MERS-coronavirus spike protein and their application in species-independent antibody detection by competitive ELISA (2018)
  • Author(s): Fukushi S, Fukuma A, Kurosu T, Watanabe S, Shimojima M, Shirato K, Iwata-Yoshikawa N, Nagata N, Ohnishi K, Ato M, Melaku SK, Sentsui H, Saijo M
  • Journal Title: J Virol Methods Volume: 251 Pages: 22-29
  • DOI: 10.1016/j.jviromet.2017.10.008
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Non Susceptibility of Neonatal and Adult Rats against the Middle East Respiratory Syndrome Coronavirus. (2016)
  • Author(s): Iwata-Yoshikawa N, Fukushi S, Fukuma A, Suzuki T, Takeda M, Tashiro M, Hasegawa H, Nagata N.
  • Journal Title: Jpn J Infect Dis. Volume: 69 Pages: 510-516
  • Peer Reviewed / Open Access
• [Journal Article] Severe Fever with Thrombocytopenia Syndrome Virus Antigen Detection Using Monoclonal Antibodies to the Nucleocapsid Protein. (2016)
  • Author(s): Fukuma A, Fukushi S, Yoshikawa T, Tani H, Taniguchi S, Kurosu T, Egawa K, Suda Y, Singh H, Nomachi T, Gokuden M, Ando K, Kida K, Kan M, Kato N, Yoshikawa A, Kitamoto H, Sato Y, Suzuki T, Hasegawa H, Morikawa S, Shimojima M, Saijo M
  • Journal Title: PLOS Neglected Tropical Diseases Volume: 10(4) Issue: 4 Pages: e0004595-e0004595
  • DOI: 10.1371/journal.pntd.0004595
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Inability of rat DPP4 to allow MERS-CoV infection revealed by using a VSV pseudotype bearing truncated MERS-CoV spike protein (2015)
  • Author(s): Aiko Fukuma, Hideki Tani, Satoshi Taniguchi, Masayuki Shimojima, Masayuki Saijo, Shuetsu Fukushi
  • Journal Title: Inability of rat DPP4 to allow MERS-CoV infection revealed by using a VSV pseudotype bearing truncated MERS-CoV spike protein. Volume: 160(9) Issue: 9 Pages: 2293-2300
  • DOI: 10.1007/s00705-015-2506-z
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] VSV pseudotype and monoclonal antibody-based assays for determining MERS coronavirus neutralizing antibody responses (2017)
  • Author(s): Shuetsu Fukushi, Aiko Fukuma, Hideki Tani, Takeshi Kurosu, Satoshi Taniguchi, Kazutaka Egawa, Masayuki Shimojima, Kazuya Shirato, Shutoku Matsuyama, Hanako Sekimukai, Naoko Iwata-Yoshikawa, Noriyo Nagata, Kazuo Ohnishi, Hiroshi Sentsui, and Masayuki Saijo
  • Organizer: XIVth International Nidovirus Symposium
  • Int'l Joint Research
• [Presentation] Characterization of novel monoclonal antibodies against MERS-coronavirus (MERS-CoV) spike protein and their application in competitive ELISA for detecting antibodies to MERS-CoV (2017)
  • Author(s): Shuetsu Fukushi, Takeshi Kurosu, Shumpei Watanabe, Masayuki Shimojima, Kazuya Shirato, Shutoku Matsuyama, Naoko Iwata-Yoshikawa, Noriyo Nagata, Kazuo Ohnishi, Manabu Ato, Hiroshi Sentsui, and Masayuki Saijo
  • Organizer: 第65回日本ウイルス学会学術集会
• [Presentation] Vesicular stomatitis virus pseudotype-based assay for detecting MERS-CoV neutralizing antibody responses. (2016)
  • Author(s): Shuetsu Fukushi, Aiko Fukuma, Hideki Tani, Takeshi Kurosu, Satoshi Taniguchi, Kazutaka Egawa, Masayuki Shimojima, Kazuya Shirato, Shutoku Matsuyama, Hiroshi Sentsui, Masayuki Saijo.
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 札幌
  • Year and Date: 2016-10-25
• [Presentation] Inability of rat DPP4 to allow MERS-CoV infection revealed by using a VSV pseudotype bearing truncated MERS-CoV spike protein (2015)
  • Author(s): Shuetsu Fukushi, Aiko Fukuma, Hideki Tani, Satoshi Taniguchi, Masayuki Shimojima, Masayuki Saijo
  • Organizer: 第６３回 日本ウイルス学会学術集会
  • Place of Presentation: 福岡
  • Year and Date: 2015-11-22
• [Presentation] Development of antigen-capture ELISA for the detection of sever fever with thrombocytopenia syndrome virus nucleoprotein (2014)
  • Author(s): Shuetsu Fukushi
  • Organizer: The 10th China-Japan International Conference of Virology
  • Place of Presentation: Changchun, China
  • Year and Date: 2014-08-25 – 2014-08-28
• [Patent(Industrial Property Rights)] MERS コロナウイルスのS 蛋白質に対するモノクローナル抗体を利用 したMERS コロナウイルス特異抗体検出法 (2017)
  • Inventor(s): 福士秀悦,大西和夫,松山州徳,白戸憲也,岩田奈織子,永田典代
  • Industrial Property Rights Holder: 国立感染症研究所
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-107132
  • Filing Date: 2017