Research on adaptive evolution and its genetic factors of Mycobacterium tuberculosis clinical strains towards development of genotyping methods

• Project/Area Number: 24689034
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Public health/Health science
• Research Institution: Nagasaki University
• Principal Investigator: WADA Takayuki 長崎大学, 熱帯医学研究所, 助教 (70332450)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2014: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2013: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2012: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000)
• Keywords: 感染症 / ゲノム / 公衆衛生 / 分子疫学 / 結核 / 社会医学 / 衛生

Outline of Final Research Achievements

To uncover uniqueness of Mycobacterium tuberculosis clinical isolates in Japan, we focused on two research targets; genetic diversities of the two unique sublineages, G3 and G4 of the Beijing lineage and complete genome sequences of representative 12 strains isolated in Japan. Various mutations of G3/G4 isolates from geographically broad areas could be determined successfully. Phylogenetic analysis based on the results shows historical dissemination of the sublineages which might cause their endemic situation in Japan. The data also reveals unreliability of genotyping methods to discriminate clinical isolates for G4 sublineage. The complete genome sequences of clinical strains showed high similarity to referential strain, H37Rv, but they can provide appropriate references to approach genetic diversity among paralogous genes.

Research Products

• [Journal Article] Clonality and micro-diversity of a nationwide spreading genotype of Mycobacterium tuberculosis in Japan. (2015)
  • Author(s): Takayuki Wada, Tomotada Iwamoto, Aki Tamaru, Junji Seto, Tadayuki, Ahiko, Kaori Yamamoto, Atushi Hase, Shinji Maeda, and Taro Yamamoto.
  • Journal Title: PLoS One Volume: 10(3) Issue: 3 Pages: e0118495-e0118495
  • DOI: 10.1371/journal.pone.0118495
  • NAID: 120006987070
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of Mycolic Acids in Total Fatty Acid Methyl Ester Fractions from <i>Mycobacterium</i> Species by High Resolution MALDI-TOFMS (2015)
  • Author(s): Teramoto K, Suga M, Sato T, Wada T, Yamamoto A, Fujiwara N
  • Journal Title: Mass Spectrometry Volume: 4 Issue: 1 Pages: A0035-A0035
  • DOI: 10.5702/massspectrometry.A0035
  • NAID: 130005095680
  • ISSN: 2186-5116, 2187-137X
  • Peer Reviewed / Open Access
• [Journal Article] Second worldwide proficiency study on variable number of tandem repeats typing of Mycobacterium tuberculosis complex (2014)
  • Author(s): de Beer JL, Kodmon C, van Ingen J, Supply P, van Soolingen D; Global Network for Molecular Surveillance of Tuberculosis 2010.
  • Journal Title: International Journal of Tuberculosis and Lung Diseases Volume: 18(5) Issue: 5 Pages: 594-600
  • DOI: 10.5588/ijtld.13.0531
  • Peer Reviewed
• [Journal Article] Pathological Findings of an outbreak of Mycobacterium marinum infection in Japanese Forest Green tree frogs, Rhacophorus arboreus (2014)
  • Author(s): Haridy M, Tachikawa Y, Yoshida S, Maeda S, Wada T, Inoue S, Sakai H, Yanai T.
  • Journal Title: Journal of Comparative Pathology Volume: In press
  • Peer Reviewed
• [Journal Article] Multiplex agarose gel electrophoresis system for variable number of tandem repeats genotyping: Analysis example using Mycobacterium tuberculosis. (2013)
  • Author(s): Wada T, Maeda S.
  • Journal Title: Electrophoresis Volume: 34 Issue: 8 Pages: 1171-1174
  • DOI: 10.1002/elps.201200471
  • Peer Reviewed
• [Journal Article] 複数自治体をまたぐ広域的結核分子疫学の基盤構築―JATA(12)-VNTR型別に基づくクラスター形成とその傾向― (2013)
  • Author(s): 和田崇之、田丸亜貴、岩本朋忠、有川健太郎、中西典子、小向潤、松本健二、長谷篤
  • Journal Title: 結核 Volume: 88 Pages: 393-398
  • NAID: 10031162884
  • Peer Reviewed
• [Journal Article] JATA(12)-VNTR型別による結核集団発生事例の菌株異同調査 (2013)
  • Author(s): 田丸亜貴、和田崇之、岩本朋忠、長谷篤
  • Journal Title: 結核 Volume: 88 Pages: 399-403
  • NAID: 10031162885
  • Peer Reviewed
• [Journal Article] Whole-genome Sequence of the Potentially Hyper-transmissible Multi-DrugResistant Mycobacterium tuberculosis Beijing Strain, OM-V02_005. (2013)
  • Author(s): Yoshitaka Tateishi, Aki Tamaru, Yoshitoshi Ogura, Mamiko Niki, TakayukiWada, Taro Yamamoto, Kazuto Hirata, Tetsuya Hayashi and Sohkichi Matsumoto.
  • Journal Title: Genome Announcement Volume: 8 Issue: 4 Pages: 608-613
  • DOI: 10.1128/genomea.00608-13
  • Peer Reviewed
• [Journal Article] 23) Evolutionary robust SNPs reveal the misclassification of Mycobacterium tuberculosis Beijing family strains into sublineages. (2013)
  • Author(s): Noriko Nakanishi, Takayuki Wada, Kentaro Arikawa, Julie Millet, Nalin Rastogi, and Tomotada Iwamoto
  • Journal Title: Infect. Genet. Evol. Volume: 16 Pages: 174-177
  • DOI: 10.1016/j.meegid.2013.02.007
  • Peer Reviewed
• [Journal Article] Dominant Incidence of Multidrug and Extensively Drug-Resistant Specific Mycobacterium tuberculosis Clones in Osaka Prefecture, Japan. (2012)
  • Author(s): Tamaru, A., C. Nakajima, T. Wada, Y. Wang, M. Inoue, R. Kawahara, R. Maekura, Y. Ozeki, H. Ogura, K. Kobayashi, Y. Suzuki, and S. Matsumoto.
  • Journal Title: PLoS One Volume: 7 Issue: 8 Pages: e42505-e42505
  • DOI: 10.1371/journal.pone.0042505
  • Peer Reviewed
• [Presentation] ゲノム解析が解き明かす結核の拡散経緯とこれから (2015)
  • Author(s): 和田 崇之
  • Organizer: 第20回国際結核セミナー
  • Place of Presentation: ヤクルトホール（東京都港区）
  • Year and Date: 2015-03-05
  • Invited
• [Presentation] Phylogenetic uniqueness and its historical backgrounds of Mycobacterium tuberculosis in Japan (2015)
  • Author(s): Wada T, Iwamoto T, Maeda S, Seto J, Tamaru A, Hase A, Murakami K, Takara T, Ahiko T, Yamamoto T.
  • Organizer: International Symposium on Genome Science 2015 "Expanding Frontiers of Genome Science II"
  • Place of Presentation: Hitotsubashi-Hall (Minato-ku, Tokyo, Japan)
  • Year and Date: 2015-01-20 – 2015-01-21
• [Presentation] 抗酸菌属に潜む分類学と臨床現場の乖離 (2015)
  • Author(s): 和田 崇之
  • Organizer: 日本分類学会連合第14回総会
  • Place of Presentation: 国立科学博物館（東京都港区）
  • Year and Date: 2015-01-11
  • Invited
• [Presentation] Opportunistic and pathogenic Mycobacteria: importance of systematic surveillance of mycobacteriosis based on pathological diagnosis (2014)
  • Author(s): Wada T, Yoshida S, Yamamoto T, Yanai T.
  • Organizer: Asian Society of Zoo and Wildlife Medicine (2014 meeting in Vietnam)
  • Place of Presentation: Tam dao National Park (Tam Dao, Vietnam)
  • Year and Date: 2014-10-14
• [Presentation] M株の広域的分離の原因究明─比較ゲノム解析に基づく「結核ゲノム疫学」の導入 (2014)
  • Author(s): 和田崇之、岩本朋忠、瀬戸順二、田丸亜貴、前田伸司、長谷篤、山本太郎
  • Organizer: 第89回日本結核病学会総会
  • Place of Presentation: 長良川国際会議場（岐阜）
  • Year and Date: 2014-05-10
• [Presentation] 日本株BCGに含まれるバリアントの比率変化の検討 (2014)
  • Author(s): 大原直也、趙娜、和田崇之、藤原永年、前田伸司、山本三郎、中山真彰、井上哲圭、瀧井猛将、前山順一
  • Organizer: 第89回日本結核病学会総会
  • Place of Presentation: 長良川国際会議場（岐阜）
  • Year and Date: 2014-05-09
• [Presentation] 超高分解能 MALDI Spiral-TOFMS によるミコール酸の簡易迅速分析法 (2014)
  • Author(s): 藤原永年、和田崇之、前田伸司
  • Organizer: 第89回日本結核病学会総会
  • Place of Presentation: 長良川国際会議場（岐阜）
  • Year and Date: 2014-05-09
• [Presentation] 結核菌国内臨床株の比較ゲノム解析：大規模拡散株と国内定着系統群を追跡する (2014)
  • Author(s): 和田 崇之
  • Organizer: 第84回実験結核研究会
  • Place of Presentation: 長良川国際会議場（岐阜）
  • Year and Date: 2014-05-08
• [Presentation] 結核菌北京型1,054株を用いた遺伝的系統解析法の評価 (2013)
  • Author(s): 中西典子、和田崇之、有川健太郎、岩本朋忠
  • Organizer: 第86回日本細菌学会総会
  • Place of Presentation: 幕張メッセ(千葉)
• [Presentation] 反復配列多型法による型別結果比較における問題点とその対策 (2012)
  • Author(s): 前田伸司、和田崇之
  • Organizer: 第87回日本結核病学会総会
  • Place of Presentation: 広島国際会議場(広島)
• [Presentation] 結核菌国内臨床株の比較ゲノム解析：大規模拡散株と国内定着系統群を追跡する
  • Author(s): 和田崇之
  • Organizer: 第84回実験結核研究会
  • Place of Presentation: 長良川国際会議場（岐阜）
• [Presentation] 結核菌における遺伝型別一致株の比較ゲノム解析
  • Author(s): 和田崇之、岩本朋忠、瀬戸順二、田丸亜貴、前田伸司、長谷篤、山本太郎
  • Organizer: 第87回日本細菌学会総会
  • Place of Presentation: タワーホール船堀（東京）
• [Presentation] 結核分子疫学研究の現在と未来
  • Author(s): 和田崇之
  • Organizer: 第19回国際結核セミナー
  • Place of Presentation: ヤクルトホール（東京）
  • Invited
• [Presentation] Pathological findings of Mycobacteriosis in Japanese forest green tree frogs, Rhacophorus arboreus.
  • Author(s): Haridy M, Tachikawa Y, Wada T, Yoshida S, Maeda S, Inoue S, Sakai H, Yanai T
  • Organizer: Combined Conference 2013, One Health in Asia Pacific.
  • Place of Presentation: Institute of Technical Education (Singapore)
• [Presentation] Mycobcterium kansasii: its epidemiological background in humans and a case of infection in black bearded Sakis in a zoo.
  • Author(s): Wada T, Yoshida S, Takata M, Tomita M, Tsuyuguchi K, Yanai T, Yamamoto T
  • Organizer: Combined Conference 2013, One Health in Asia Pacific.
  • Place of Presentation: Institute of Technical Education (Singapore)
• [Presentation] 反復配列多型(VNTR)分析法による結核菌型別―精度管理と考え方,使い方―
  • Author(s): 和田崇之
  • Organizer: 第72回日本公衆衛生学会総会
  • Place of Presentation: 三重県総合文化センター（三重）
  • Invited
• [Presentation] わが国における結核分子疫学と伝搬経路推定:結核中蔓延国としての役割について
  • Author(s): 和田崇之，山本太郎
  • Organizer: 第54回日本熱帯医学会大会
  • Place of Presentation: 長崎ブリックホール（長崎）
• [Presentation] Genotyping of Mycobacterium tuberculosis (MTB), especially Beijing genotype MTB, in Japan by using the variable-number of tandem repeats (VNTR) and a next generation sequencer (NGS).
  • Author(s): Maeda S, Wada T, Naka T, Shibata M, Fujiwara N
  • Organizer: The 5th EMBO meeting 2013
  • Place of Presentation: Amsterdam RAI (Netherlands)
• [Presentation] 結核菌の遺伝的多様性がもたらす研究展開：公衆衛生から自然史まで
  • Author(s): 和田崇之
  • Organizer: H25年度長崎県職臨床技師会研修会
  • Place of Presentation: 長崎県央保健所（長崎）
  • Invited
• [Remarks] 結核菌VNTRハンドブック
  • URL: http://www.chieiken.gr.jp/