You Chunlin, Hamasuna Ryoichi, Ogawa Midori, Fukuda Kazumasa, Hachisuga Toru, Matsumoto Tetsuro, Taniguchi Hatsumi
Department of Microbiology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.
Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.
Microb Pathog. 2016 Jun;95:95-100. doi: 10.1016/j.micpath.2016.02.022. Epub 2016 Mar 22.
To analyse the bacterial flora of urine from patients with male urethritis using the clone library method.
Urine specimens from patients with urethritis were used. The bacterial flora was analysed according to the 16S ribosomal RNA gene-based clone library method. In addition, Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum or Ureaplasma parvum were detected by the conventional PCR methods (TMA or real-time PCR) and data from the clone library and conventional PCR methods were compared.
Among 58 urine specimens, 38 were successfully analysed using the clone library method. From the specimens, 2427 clones were evaluated and 95 bacterial phylotypes were detected. N. gonorrhoeae was detected from 6 specimens and as the predominant bacterial species in 5 specimens. M. genitalium was detected from 5 specimens and as the predominant bacterial species in 3 specimens. C. trachomatis was detected from 15 specimens using the TMA method, but was detected from only 1 specimen using the clone library method. U. parvum was detected from only 2 specimens using the clone library method. In addition, Haemophilus influenzae and Neisseria meningitidis were also detected in 8 and 1 specimens, respectively. Gardnerella vaginalis, which is a potential pathogen for bacterial vaginitis in women, was detected in 10 specimens.
The clone library method can detect the occupancy rate of each bacteria species among the bacterial flora and may be a new method for bacterial analyses in male urethritis.
采用克隆文库法分析男性尿道炎患者尿液中的细菌菌群。
使用尿道炎患者的尿液标本。根据基于16S核糖体RNA基因的克隆文库法分析细菌菌群。此外,通过传统PCR方法(转录介导扩增法或实时PCR)检测淋病奈瑟菌、沙眼衣原体、生殖支原体、人型支原体、解脲脲原体或微小脲原体,并比较克隆文库法和传统PCR方法的数据。
在58份尿液标本中,38份成功采用克隆文库法进行分析。从这些标本中,评估了2427个克隆,检测到95种细菌系统型。6份标本中检测到淋病奈瑟菌,其中5份标本中该菌为主要细菌种类。5份标本中检测到生殖支原体,其中3份标本中该菌为主要细菌种类。采用转录介导扩增法从15份标本中检测到沙眼衣原体,但采用克隆文库法仅从1份标本中检测到。采用克隆文库法仅从2份标本中检测到微小脲原体。此外,分别在8份和1份标本中检测到流感嗜血杆菌和脑膜炎奈瑟菌。在10份标本中检测到阴道加德纳菌,该菌是女性细菌性阴道炎的潜在病原体。
克隆文库法可检测细菌菌群中各细菌种类的占有率,可能是男性尿道炎细菌分析的一种新方法。