Molecular analysis of small nucleic acid molecules for the development of novel diagnostic methods of canine leishmaniasis

• Project/Area Number: 16H05023
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Hokkaido University
• Principal Investigator: KATAKURA KEN 北海道大学, 獣医学研究院, 特任教授 (10130155)
• Research Collaborator: NAKAO Ryo 北海道大学, (連合)獣医学研究院, 准教授 (50633955)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥16,770,000 (Direct Cost: ¥12,900,000、Indirect Cost: ¥3,870,000); Fiscal Year 2018: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2017: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2016: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
• Keywords: イヌ・リーシュマニア症 / 人獣共通感染症 / エキソソームRNA / セルフリーDNA / 早期血清診断法 / 次世代診断法 / 低分子核酸 / 次世代シーケンス / 犬リーシュマニア症

Outline of Final Research Achievements

Canine leishmaniasis is a chronic disease caused by infections with several Leishmania species. This study aims to develop more sensitive novel diagnostic methods of leishmaniasis based on analysis of small nucleic acids such as cell-free DNA and small RNA molecules in the peripheral blood. Cell-free DNAs of possible Leishmania-origin were detected by next generation sequence analysis from the blood of Leishmania donovani-infected stray dogs captured in the endemic area of human visceral leishmaniasis in Bangladesh. Meanwhile, small RNA molecules derived from possible Leishmania rRNAs and tRNAs were detected in the blood of mice 2 weeks after experimental infection with Leishmania donovani. These results suggest that small nucleic acids such as cell-free DNA and small RNA molecules are useful for novel diagnostic molecular markers of leishmaniasis.

Academic Significance and Societal Importance of the Research Achievements

この研究は、リーシュマニアという寄生虫に感染しているかどうかを、できるだけ早期にそして正確に判断するために、より精度の高い診断法を開発することを目的として行われました。リーシュマニアに感染した犬やマウスの血液を調べたところ、リーシュマニア由来の小さな核酸分子が、感染動物の血液中にごく微量含まれていることが分かりました。用いた次世代シーケンスデータ解析技術は、１つの検体から様々な情報を得ることができる技術です。今回、この方法が寄生虫感染の診断にも応用できることを実証することができました。この結果は、今後のさらなる診断技術の開発・改良に貢献するものです。

Research Products

• [Int'l Joint Research] Bangladesh Agricultural University(バングラデシュ)
• [Journal Article] Potential role of dogs as sentinels and reservoirs for piroplasms infecting equine and cattle in Riyadh City, Saudi Arabia (2019)
  • Author(s): Salim Bashir、Alanazi Abdullah D.、Omori Ryosuke、Alyousif Mohamed S.、Alanazi Ibrahim O.、Katakura Ken、Nakao Ryo
  • Journal Title: Acta Tropica Volume: 193 Pages: 78-83
  • DOI: 10.1016/j.actatropica.2019.02.029
  • NAID: 120006843068
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tick-borne haemoparasites and Anaplasmataceae in domestic dogs in Zambia (2018)
  • Author(s): Qiu Yongjin、Kaneko Chiho、Kajihara Masahiro、Ngonda Saasa、Simulundu Edgar、Muleya Walter、Thu May J、Hang’ombe Bernard、Katakura Ken、Takada Ayato、Sawa Hirofumi、Simuunza Martin、Nakao Ryo
  • Journal Title: Ticks and Tick-borne Diseases Volume: 印刷中 Issue: 4 Pages: 988-995
  • DOI: 10.1016/j.ttbdis.2018.03.025
  • NAID: 120006626572
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Potential of cell-free DNA as a screening marker for parasite infections in dog (2018)
  • Author(s): Akter Shirin、Nakao Ryo、Imasato Yuhei、Alam Mohammad Zahangir、Katakura Ken
  • Journal Title: Genomics Volume: - Issue: 4 Pages: 906-912
  • DOI: 10.1016/j.ygeno.2018.05.020
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular and serological evidence of Leishmania infection in stray dogs from visceral leishmaniasis-endemic areas of Bangladesh. (2016)
  • Author(s): Akter S, Alam MZ, Nakao R, Yasin MG, Kato H, Katakura K.
  • Journal Title: Am J Trop Med Hyg. Volume: 95 Issue: 4 Pages: 795-799
  • DOI: 10.4269/ajtmh.16-0151
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] An experimental challenge model of visceral leishmaniasis by Leishmania donovani promastigotes in mice. (2016)
  • Author(s): Katakura K
  • Journal Title: Parasitol Int Volume: 65 Issue: 5 Pages: 603-606
  • DOI: 10.1016/j.parint.2016.03.008
  • Peer Reviewed / Int'l Joint Research
• [Presentation] PCR detection and phylogenetic analysis of haemoprotozoan parasites in livestock in Myanmar by sequence analysis of selected target genes (2018)
  • Author(s): Ken Katakura, Saw Bawm, Hla Myet Chel, May June Thu, Lat Lat Htun, Yuhei Imasato, Ryo Nakao
  • Organizer: 14th International Conference on Molecular Epidemiology and Evolutionary Genetics of Infectious Diseases (MEEGID XIV)
  • Int'l Joint Research
• [Presentation] Molecular and serological evidence of Leishmania infection in stray dogs from visceral leishmaniasis endemic areas of Bangladesh (2017)
  • Author(s): Akter S, Nakao R, Alam MZ, Yasin G, Katakura K
  • Organizer: 6th World Congress on Leishmaniasis (Worldleish-6)
  • Int'l Joint Research
• [Presentation] Potential of cell-free DNA as a novel diagnostic biomarker for parasite infections in dog (2017)
  • Author(s): Akter S, Nakao R, Katakura K
  • Organizer: 26th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP)
  • Int'l Joint Research
• [Presentation] Molecular and serological evidence of Leishmania infection in stray dogs from visceral leishmaniasis endemic areas of Bangladesh. (2016)
  • Author(s): Shirin Akter, 中尾亮、Mohammad Zahangir Alam, Golam Yasin, 片倉 賢
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 日本大学（藤沢市）
  • Year and Date: 2016-09-06
• [Book] Echinococcosis (2017)
  • Author(s): Enkai S, Sakamoto K, Kaneko M, Kouguchi H, Irie T, Yagi K, Ishida Y, Matsumoto J, Oku Y, Katakura K, Fujita O, Nozaki T, Kita K
  • Publisher: InTechOpen
  • ISBN: 9789535135920
• [Remarks] 北海道大学大学院獣医学研究院・獣医学部 獣医学大学院研究室一覧 寄生虫学教室
  • URL: https://www.vetmed.hokudai.ac.jp/research/detail/parasitology/
• [Remarks] 北海道大学大学院獣医学研究院・獣医学部 寄生虫学教室ホームページ
  • URL: https://www.vetmed.hokudai.ac.jp/organization/parasitol/index.html
• [Remarks] 北海道大学大学院獣医学研究院・獣医学部 寄生虫学教室のHP
  • URL: http://www.vetmed.hokudai.ac.jp/organization/parasitol/index.html