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Detection of gene mutations in kanamycin-resistant Mycobacterium tuberculosis and development of the method for rapid diagnosis

Research Project

Project/Area Number 08670470
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Public health/Health science
Research InstitutionOsaka Prefectural Institute of Public Health

Principal Investigator

KATSUKAWA Chihiro  Osaka Prefectural Institute of Public Health, Department of Public Health, Senior Research Scientist, 公衆衛生部, 主任研究員 (20183725)

Co-Investigator(Kenkyū-buntansha) TAMARU Aki  Osaka Prefectural Institute of Public Health, Department of Public Health, Resea, 公衆衛生部, 研究員 (70270767)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
KeywordsMycobacterium tuberculosis / kanamycin / kanamycin-resistant strains / rrs gene / PCR-RFLP analysis / efg遺伝子 / rpsG遺伝子
Research Abstract

In Mycobacterium smegmatis and a limited number of Mycobacterium tuberculosis strains, the involvement of alterations of the 16S rRNA gene (rrs) in resistance to kanamycin has been shown. To investigate the extent to which mutations in a specific region of the n-s gene and the kanamycin-resistant phenotype in clinically isolated M.tuberculosis swains were correlated, 43 kanamycin-resistant swains (MICs, >=200 mu g/ml), 71 kanamycin-susceptible strains, and 4 type strains were examined. The 300-bp DNA fragments carrying the rrs gene and the intervening sequence between the rrs gene and 23S rRNA (rrl) gene fragments were amplified by PCR and were subjected to PCR-based direct sequencing. By comparing the nucleotide sequences, substitutions were found in 29 of 43 (67.4%) kanamycin-resistant clinical isolates at position 1400, 1401 and 1483 but in none of the 71 sensitive isolates or the 4 type strains. The most frequent substitution, from A to G, occurred at position 1400. A substitution from C to T at position 1401 was found once. Two clinical isolates carried the double mutation from C to A at position 1401 and from G to T at position 1483. In addition, we found that these mutations can be distinguished from wild-type strains by digestion with the restriction endonucleases Tai I and Tsp 451. Furthermore, we found that the genotypes of kanamycin-resistant swains can be discriminated from each other by digestion with a restriction endonuclease, Bst UI or Dde I.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Yasuhiko Suzuki: "Detection of kanamycin-resistant Mycobacterium tuberculosis by identifying mutations in the 16S rRNA gene" Journal of Clinical Microbiology. 36. 1220-1225 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Chihiro Katsukawa: "Characterization of the rpsL and rrs genes of streptomycin-resistant clinical isolates of Mycobacterium tuberculosis in Japan." Journal of Applied Microbiology. 83. 634-640 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasuhiko Suzuki: "Immunological characterization of five recombinant Mycobacterium leprae proteins produced in Escherichia coli" Proceeding of 33rd US-Japan Tuberculosis and Leprosy Symposium. 33. 58-61 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Aki Tamaru: "Detection of mutations on the genome of drug resistant clinical isolates of Mycobacterium tuberculosis in Japan" Proceeding of 33rd US-Japan Tuberculosis and Leprosy Symposium. 33. 190-190 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 鈴木定彦: "結核菌の薬剤耐性検査 薬剤耐性に関与する遺伝子と迅速診断への応用の可能性" 臨床と微生物. 25. 795-801 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 勝川千尋: "結核の現状" 防菌防黴. 27. 315-335 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasuhiko Suzuki, Chihiro Katsukawa, Aki Tamaru, Chiyoji Abe, Masanao Makino, Yasuo Mizuguchi, and Hatsumi, Taniguchi: "Detection of kanamycin-resistan Mycobacterium tuberculosis by identifyingmutations in the 16S rRNA gene." J.Clin.Microbiol.36. 1220-1225 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasuhiko Suzuki, Chihiro Katsukawa, Aki Tamaru, and Masanao Makino: "Application of recombinant 85 complex of Mycobacterium leprae for sero-diagnosis of leprosy : Over expression of MPT51 like protein(MPT51) of M.leprae in Escherichia coli." Proceeding of 31st US-Japan Tuberculosis and Leprosy Symposium.76-80 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Chihiro Katsukawa, Aki Tamaru, Yoshihito Miyata, Chiyoji Abe, Masanao Makino, and Yasuhiko Suzuki: "Characterization of the rpsL and rrs genes of streptomycin-resistant clinical isolate of Mycobacterium tuberculosis in Japan." J.Appl.Microbiol.83. 634-640 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasuhiko Suzuki, Chihiro Katsukawa, Aki Tamaru, Masanao Makino, and Masanori Matsuoka: "immunological characterization of five recombinant Mycobacterium leprae proteins produced in Escherichia coli." proceeding of 33rd US-Japan Tuberculosis and Leprosy Symposium. 58-61 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Aki Tamaru, Chihiro Katsukawa, Yasuhiko Suzuki, Chiyoji Abe, Masanao Makino, Yasuo Mizuguchi, and Hatsumi Taniguchi: "Detection of mutations on the genome of drug resistant clinical isolates of Mycobacterium tuberculosis in Japan." Proceeding of 33rd US-Japan Tuberculosis and Leprosy Symposium. 190 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Chihiro Katsukawa: "A comprehensive report of the state of tuberculosis in Japan." Journal of Antibacterial and Antifungal Agents. 27. 315-335 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasuhiko Suzuki: "Detection of kanamycin-resistant Mycobacterium tuberculosis identifying mutations in the 16S rRNA gene" Journal of Clinical Microbiology. 36. 1220-1225 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Chihiro Katsukawa: "A comprebensive report of the state of tuberculosis in Japan" Journal of Antibacterial and Antifungal Agents. 27. 315-335 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yasuhiko Suzuki: "Immunological characterization of five recombinant Mycobacterium leprae proteins produced in Escherichia coli" Proceeding of 33rd US-Japan Tuberculosis and Leprosy Symposium. 33. 58-61 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Aki Tamaru: "Detection of mulations on the genome of drug resistant clinical isolates of Mycobacterium tuberculosis in Japan" Proceeding of 33rd US-Japan Tuberculosis and Leprosy Symposium. 33. 190-190 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 鈴木定彦: "結核菌の薬剤耐性検査 薬剤耐性に関与する遺伝子と迅速診断への応用の可能性" 臨床と微生物. 25. 795-801 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Chihiro Katsukawa,Aki Tamaruet al.: "Characterization of the rpsL and rrs genes of streptomycin-resistant clinical isolates of Mycobacterium tuberculosis in Japan." Jouranl of Applied Microbiology.83. 634-640 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yoshishige Inagaki,Chihiro Katsukawa et al.: "Serotyping of Streptococcus pyogenes isolated from common and severe invasive infections in Japan,1990-5:implication of the T3 serotype strain-expansion in TSLS." Epidemiology and Infection.119. 41-48 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yasuhiko Suzuki,Chihiro Katsukawa,Aki Tamaru and Masanao Makino: "Application of Recombinant 85 Complex of Nycobacterium leprae for serodiagnosis of Leprosy : Over expression of MPT51 like protein (MPL51) of M.leprae in Escherichia coli." Thirty-First Research Conference on Tuberculosis and Leprosy. The U.S.-Japan Cooperative Medical Science Program.31. 76-80 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kyongsu Hong and Chihiro Katsukawa: "Expression and Characterization of a Recombinant M3 Ploypeptide encoding Amino acids 42 through 517 of M3 protein" Immunology and Infectious Diseases. 6. 207-214 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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