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Selective whole genome amplification for mitogenome analysis of disease vectors

Research Project

Project/Area Number 20K21358
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 42:Veterinary medical science, animal science, and related fields
Research InstitutionHokkaido University

Principal Investigator

Nakao Ryo  北海道大学, 獣医学研究院, 准教授 (50633955)

Project Period (FY) 2020-07-30 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2021: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2020: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Keywords選択的ゲノム増幅法 / 節足動物 / マダニ / ミトゲノム / SWGA
Outline of Research at the Start

マダニや蚊などの節足動物は様々な病原体を保有し人や動物に伝播する。節足動物によって媒介される感染症を制御するためには、節足動物を遺伝的に同定でき、さらにその集団遺伝構造を解析する手法が有用である。本研究では節足動物のミトコンドリアゲノム(ミトゲノム)配列を簡便かつ安価に解読できる手法の開発に挑戦する。ミトゲノム配列情報を活用することにより、病原体の保有・伝播に関わる節足動物側素因の解析や病原体を保有する節足動物集団の分布解析が可能となる。

Outline of Final Research Achievements

The purpose of this study was to develop a method to sequence complete mitochondrial genomes (mitogenomes) of ticks by using a selective whole genome amplification (SWGA) method. The tick mitogenome-specific primers were selected based on the alignment of tick mitogenome sequences available in the public database. The tick DNA was incubated with specific primers and phi29 DNA polymerase and the resulting products were sequenced on a MiSeq platform. The results indicated that SWGA could increase the relative abundance of mitochondrial DNA by more than 10,000 holds in comparison to chromosomal DNA. The developed method enables us to sequence complete tick mitogenomes in an easy and low-cost way. We proved that the method is applicable to many tick species, of which mitogenome sequences have not yet been reported.

Academic Significance and Societal Importance of the Research Achievements

開発した選択的ミトゲノム増幅技術により、簡便かつ安価にマダニのミトゲノム情報を取得することができることとなった。このことにより、世界中でマダニのミトゲノムの解読が進み、マダニの系統進化、集団遺伝構造を理解するための遺伝子情報の集積が期待できる。また、得られたゲノム情報を活用することで、精度の高いマダニの遺伝子同定技術の開発が可能となり、マダニが媒介する感染症の対策に貢献できる。また、本研究で確立した手法は、他の生物群にも応用可能であり、生物を対象とした広い研究分野への波及効果が期待できる。

Report

(3 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • Research Products

    (24 results)

All 2022 2021 2020 Other

All Int'l Joint Research (11 results) Journal Article (11 results) (of which Int'l Joint Research: 11 results,  Peer Reviewed: 11 results,  Open Access: 9 results) Presentation (2 results)

  • [Int'l Joint Research] サウスバリー大学/マンスーラ大学(エジプト)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] リロングウェ農業天然資源大学(マラウイ)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] ザンビア大学(ザンビア)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] クイーンズランド大学(オーストラリア)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] ハルツーム大学(スーダン)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] サウスバリー大学/マンスーラ大学(エジプト)

    • Related Report
      2020 Research-status Report
  • [Int'l Joint Research] リロングウェ農業天然資源大学(マラウイ)

    • Related Report
      2020 Research-status Report
  • [Int'l Joint Research] ザンビア大学(ザンビア)

    • Related Report
      2020 Research-status Report
  • [Int'l Joint Research] クイーンズランド大学(オーストラリア)

    • Related Report
      2020 Research-status Report
  • [Int'l Joint Research] ハルツーム大学(スーダン)

    • Related Report
      2020 Research-status Report
  • [Int'l Joint Research]

    • Related Report
      2020 Research-status Report
  • [Journal Article] Reconstruction of mitochondrial genomes from raw sequencing data provides insights on the phylogeny of Ixodes ticks and cautions for species misidentification2022

    • Author(s)
      Mohamed Wessam Mohamed Ahmed、Moustafa Mohamed Abdallah Mohamed、Kelava Samuel、Barker Dayana、Matsuno Keita、Nonaka Nariaki、Shao Renfu、Mans Ben J.、Barker Stephen C.、Nakao Ryo
    • Journal Title

      Ticks and Tick-borne Diseases

      Volume: 13 Issue: 1 Pages: 101832-101832

    • DOI

      10.1016/j.ttbdis.2021.101832

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Description of the female, nymph and larva and mitochondrial genome, and redescription of the male of Ixodes barkeri Barker, 2019 (Acari: Ixodidae), from the short-beaked echidna, Tachyglossus aculeatus, with a consideration of the most suitable subgenus for this tick2022

    • Author(s)
      Barker Dayana、Kelava Samuel、Shao Renfu、Seeman Owen D.、Jones Malcolm K.、Nakao Ryo、Barker Stephen C.、Apanaskevich Dmitry A.
    • Journal Title

      Parasites & Vectors

      Volume: 15 Issue: 1 Pages: 117-117

    • DOI

      10.1186/s13071-022-05165-2

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] High infection rate of tick-borne protozoan and rickettsial pathogens of cattle in Malawi and the development of a multiplex PCR for Babesia and Theileria species identification2022

    • Author(s)
      Chatanga Elisha、Maganga Emmanuel、Mohamed Wessam Mohamed Ahmed、Ogata Shohei、Pandey Gita Sadaula、Abdelbaset Abdelbaset Eweda、Hayashida Kyoko、Sugimoto Chihiro、Katakura Ken、Nonaka Nariaki、Nakao Ryo
    • Journal Title

      Acta Tropica

      Volume: 231 Pages: 106413-106413

    • DOI

      10.1016/j.actatropica.2022.106413

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Rediscovery of Ixodes confusus in Australia with the first description of the male from Australia, a redescription of the female and the mitochondrial (mt) genomes of five species of Ixodes2022

    • Author(s)
      Barker Dayana、Kelava Samuel、Seeman Owen D.、Shao Renfu、Seaniger James R.、Jones Malcolm K.、Apanaskevich Maria A.、Nakao Ryo、Apanaskevich Dmitry A.、Barker Stephen C.
    • Journal Title

      International Journal for Parasitology: Parasites and Wildlife

      Volume: 18 Pages: 1-11

    • DOI

      10.1016/j.ijppaw.2022.03.006

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Screening of tick-borne pathogens in argasid ticks in Zambia: Expansion of the geographic distribution of Rickettsia lusitaniae and Rickettsia hoogstraalii and detection of putative novel Anaplasma species.2021

    • Author(s)
      Qiu Y, Simuunza M, Kajihara M, Chambaro H, Harima H, Eto Y, Simulundu E, Squarre D, Torii S, Takada A, Hangmbe BM, Sawa H, Sugimoto C, Nakao R
    • Journal Title

      Ticks Tick Borne Dis

      Volume: 12 Issue: 4 Pages: 101720-101720

    • DOI

      10.1016/j.ttbdis.2021.101720

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Applications of Blocker Nucleic Acids and Non-Metazoan PCR Improves the Discovery of the Eukaryotic Microbiome in Ticks2021

    • Author(s)
      Taya Yurie、Kinoshita Gohta、Mohamed Wessam Mohamed Ahmed、Moustafa Mohamed Abdallah Mohamed、Ogata Shohei、Chatanga Elisha、Ohari Yuma、Kusakisako Kodai、Matsuno Keita、Nonaka Nariaki、Nakao Ryo
    • Journal Title

      Microorganisms

      Volume: 9 Issue: 5 Pages: 1051-1051

    • DOI

      10.3390/microorganisms9051051

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Molecular Detection and Genotyping of Coxiella-Like Endosymbionts in Ticks Collected from Animals and Vegetation in Zambia2021

    • Author(s)
      Kobayashi Toshiya、Chatanga Elisha、Qiu Yongjin、Simuunza Martin、Kajihara Masahiro、Hang’ombe Bernard Mudenda、Eto Yoshiki、Saasa Ngonda、Mori-Kajihara Akina、Simulundu Edgar、Takada Ayato、Sawa Hirofumi、Katakura Ken、Nonaka Nariaki、Nakao Ryo
    • Journal Title

      Pathogens

      Volume: 10 Issue: 6 Pages: 779-779

    • DOI

      10.3390/pathogens10060779

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Molecular Survey of Babesia and Anaplasma Infection in Cattle in Bolivia2021

    • Author(s)
      Ogata Shohei、Pereira Juan Antonio Cristian、Jhonny Loza Vega Ariel、Carolina Herbas Perez Gladys、Matsuno Keita、Orba Yasuko、Sawa Hirofumi、Kawamori Fumihiko、Nonaka Nariaki、Nakao Ryo
    • Journal Title

      Veterinary Sciences

      Volume: 8 Issue: 9 Pages: 188-188

    • DOI

      10.3390/vetsci8090188

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Spiroplasma Infection among Ixodid Ticks Exhibits Species Dependence and Suggests a Vertical Pattern of Transmission2021

    • Author(s)
      Ogata Shohei、Mohamed Wessam Mohamed Ahmed、Kusakisako Kodai、Thu May June、Qiu Yongjin、Moustafa Mohamed Abdallah Mohamed、Matsuno Keita、Katakura Ken、Nonaka Nariaki、Nakao Ryo
    • Journal Title

      Microorganisms

      Volume: 9 Issue: 2 Pages: 333-333

    • DOI

      10.3390/microorganisms9020333

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Exploring Prokaryotic and Eukaryotic Microbiomes Helps in Detecting Tick-Borne Infectious Agents in the Blood of Camels2021

    • Author(s)
      Mohamed Wessam Mohamed Ahmed、Ali Alsagher O.、Mahmoud Hassan Y. A. H.、Omar Mosaab A.、Chatanga Elisha、Salim Bashir、Naguib Doaa、Anders Jason L.、Nonaka Nariaki、Moustafa Mohamed Abdallah Mohamed、Nakao Ryo
    • Journal Title

      Pathogens

      Volume: 10 Issue: 3 Pages: 351-351

    • DOI

      10.3390/pathogens10030351

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Molecular detection and characterization of tick-borne hemoparasites and Anaplasmataceae in dogs in major cities of Malawi2020

    • Author(s)
      Chatanga Elisha、Kainga Henson、Razemba Tinotenda、Ssuna Richard、Swennen Lieza、Hayashida Kyoko、Sugimoto Chihiro、Katakura Ken、Nonaka Nariaki、Nakao Ryo
    • Journal Title

      Parasitology Research

      Volume: 120 Issue: 1 Pages: 267-276

    • DOI

      10.1007/s00436-020-06967-y

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] ミトゲノムおよびゲノムワイド一塩基多型を用いたフタトゲチマダニの分子遺伝学的解析2021

    • Author(s)
      尾針由真、Wessam Mohamed Ahmed Mohamed、Mohamed Abdallah Mohamed Moustafa、Elisha Chatanga、松野啓太、Stephen Barker、野中成晃、中尾亮
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Application of selective whole genome amplification (SWGA) for tick mitochondrial genome sequencing2021

    • Author(s)
      Elisha Chatanga, Tateki Matsumoto, Mohamed Abdallah Mohamed Moustafa, Yuma Ohari, Yongjin Qiu, Ken Katakura, Nariaki Nonaka, Ryo Nakao
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report

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Published: 2020-08-03   Modified: 2023-01-30  

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