|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)
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.