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Development of DNA microarry-based molecular diagnosis system for male urethritis

Research Project

Project/Area Number 13671643
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Urology
Research InstitutionGIFU UNIVERSITY

Principal Investigator

DEGUCHI Takashi  SCHOOL OF MEDICINE, PROFESSOR, 医学部, 教授 (40163935)

Co-Investigator(Kenkyū-buntansha) YOKOI Shigeki  UNIVERSITY HOSPITAL, ASSISTANT PROFESSOR, 医学部附属病院, 助手 (20313900)
ISHIHARA Satoshi  SCHOOL OF MEDICINE, ASSOCIATE PROFESSOR, 医学部, 助教授 (00193342)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2001: ¥2,600,000 (Direct Cost: ¥2,600,000)
KeywordsUrethritis / PCR / Microarray / Mycoplasma / Ureaplasma / DNAチップ / 淋菌 / クラミジア
Research Abstract

In order to detect pathogens causing non-chlamydial non-gonococcal urethritis (NCNGU), we developed a molecular diagnosis system. First, we amplified the 16s rRNA genes from clinical specimens of men with NCNGU by the universal primers for mycoplasmas and ureaplasmas, and then sequenced the PCR products. We identified the pathogen based on the sequence of the amplified 16s rRNA gene. We detected only Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum and Ureaplasma parvum in the clinical specimens. Second, we fixed species-specific oligonucleotides of M. genitalium, M. hominis, U. urealyticum and U. parvum on the microtiter plate, and hybridized the PGR products with each of the oligonucleotides on the microtiter plate. Using this assay, we could identify only M. genitalium, M. hominis, U. urealyticum and U. parvum in the clinical specimens. Now we are preparing DNA microarrays, on which the DNAs of Neisseria gonorrohoeae, Chlamydia trachomatis, Bacteroides ureolyticum, Haemphilus influenzae, Haemphilus parainfluenzae, and other bacteria are fixed. Third, we developed a quantitative assay for M. genitalium. We determined the bacterial loads of the mycoplasma in the first-pass urine of men with NCNGU, and demonstrated that the bacterial loads were significantly related to the presence of urethritis symptoms and inflammatory signs. We are also developing a quantitative assay for U. urealyticum.
We aim for the establishment of a comprehensive diagnosis system for urethritis and for the quantitative assessment of bacterial loads in men with urethritis.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Maeda, S et al.: "Association of persistence of Mycoplamsa genitalium in the urethra with recurrence of nongonococcal urethritis"Sxually Transmitted Diseases. 28(8). 472-476 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Deguchi, T et al.: "Analysis of the gyrA and parC genes of Mycoplasma genitalium detected in first-pass urine of men with non-gonococcal urethritis before and after fluoroquinolone treatment"Journal of Antimicrobial Chemotherapy. 48(11). 742-744 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Deguchi, T et al.: "Mycoplasma genitalium, another important pathogen of non-gonococcal urethritis"The Journal of Urology. 167. 1210-1217 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida, T et al.: "Phylogeny-based rapid identification of mycoplasmas and ureaplasmas from urethritis patinets"Journal of Clinical Microbiology. 40(1). 105-110 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Li, Z et al.: "Rapid detection of quinolone resistance-associated gyrA mutations in Neisseria gonorrhoeae with a Light Cycler"Journal of Infection and Chemotherapy. 9(2). 145-150 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida, T et al.: "Quantitative detection of Mycoplasma genitalium from first-pass urine of men with urethritis and asymptomatic men by real-time PCR"Journal of Clinical Microbiology. 40(4). 1451-1455 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Deguchi, T et al.: "Longitudinal Quantitative Detection of Mycoplasma genitalium in First-Pass Urine of Men with Recurrent Non-Gonococcal Urethritis by Real-Time PCR"Journal of Clinical Microbiology. 40(10). 3854-3856 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida, T et al.: "Rapid Detection of Mycoplasma genitalium, Mycoplasuma hominis, Ureaplasma parvum and Ureaplasma urealyticum in Genitourinary Samples by PCR-Microtiter Plate Hybridization Assay"Journal of Clinical Microbiology. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Maeda, S., M. Tamaki, K. Kojima, T. Yoshida, H. Ishiko, M. Yasuda, T. Deguchi: "Association of persistence of Mycoplasma genitalium in the urethra with recurrence of nongonococcal urethritis"Sexually Transmitted Diseases. 28 (8). 472-476 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Deguchi, T., S-I. Maeda, M. Tamaki, T. Yoshida, H. Ishiko, M. Ito, S. Yokoi, Y. Takahashi and S. Ishihara: "Analysis of the gyrA and parC genes of Mycoplasma genitalium detected in first-pass urine of men with non-gonococcal urethritis before and after fluoroquinolone treatment"Journal of Antimicrobial Chemotherapy. 48 (11). 742-744 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Deguchi T., S-I. Maeda: "Mycoplasma genitalium, another important pathogen of non-gonococcal urethritis"The Journal of Urology. 167. 1210-1217 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida, T., S-I. Maeda T. Deguchi and H. Ishiko: "Phylogeny-based rapid identification of mycoplasmas and ureaplasmas from urethritis patinets"Journal of Clinical Microbiology. 40 (1). 105-110 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Li, Z., S. Yokoi, Y. Kawamura, S-I. Maeda, T. Ezaki, T. Deguchi: "Rapid detection of quinolone resistance-associated gyrA mutations in Neisseria gonorrhoeae with a Light Cycler."Journal of Infection and Chemotherapy. 9 (2). 145-150 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida, T., T. Deguchi, M. Ito, S-I. Maeda, M. Tamaki, H. Ishiko: "Quantitative detection of Mycoplasma genitalium from first-pass urine of men with urethritis and asymptomatic men by real-time PCR."Journal of Clinical Microbiology. 40 (4). 1451-1455 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Deguchi, T., T. Yoshida, S. Yokoi, M. Ito, M. Tamaki, H. Ishiko, S. Maeda: "Longitudinal Quantitative Detection of Mycoplasma genitalium in First-Pass Urine of Men with Recurrent Non-Gonococcal Urethritis by Real-Time PCR."Journal of Clinical Microbiology. 40 (10). 3854-3856 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida, T., S. Maeda, T. Deguchi, T. Miyazawa, H. Ishiko: "Rapid Detection of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum and Ureaplasma urealyticum in Genitourinary Samples by PCR-Microtiter Plate Hybridization Assay."Journal of Clinical Microbiology. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takashi Yoshida et al.: "Quantitative detection of mycoplasma genitalium from first-pass urine of men with urethritis and asymptomatic men by real-time PCR"Journal of Clinical Microbiology. 40(5). 1451-1455 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Takashi Deguchi et al.: "Longitudinal Quantitative Detection of Mycoplasma genitalium in First-Pass Urine of Men with Recurrent Non-Gonococcal Urethritis by Real-Time PCR"Journal of Clinical Microbiology. 40(10). 3854-3856 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Takashi Yoshida et al.: "Rapid Detection of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum in Genitourinary Samples by PCR-Microtiter Plate Hybridization Assay"Journal of Clinical Microbiology. (in press).

    • Related Report
      2002 Annual Research Report
  • [Publications] Takashi Deguchi et al.: "Analysis of the gyrA and parC genes of Mycoplasma genitalium detected in first-pass urine of men with non-gonococcal urethritis before and after fluoroquinolone treatment"Journal of Antimicrobial Chemotherapy. 48(11). 742-744 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] SHIN-ICHI MAEDA et al.: "Association of persistence of Mycoplasma genitalium in the urethra with recurrence of nongonococcal urethritis"Sexually Transmitted Diseases. 28(8). 472-476 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takashi Yoshida et al.: "Phylogeny-based rapid identification of mycoplasmas and ureaplasmas from urethritis patinets"Journal of Clinical Microbiology. 40(1). 105-110 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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