1887

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Volume 58, Issue 7

Real-time PCR detection of the gene of unknown function of in men with and without non-gonococcal urethritis and their female partners in England Free

Abstract

Real-time PCR was employed to detect a region of the gene, a gene of unknown function, in clinical samples. Amplification of DNA and signal production from 15 other species of human mycoplasmas and 14 other bacteria and viruses did not occur. Using a panel of 208 genital and rectal samples, the sensitivity when compared to the modified gene (encoding the major surface protein MgPa) real-time PCR assay was found to be 100 % and the specificity of the assay 99.5 % with a positive predictive value of 80 % and a negative predictive value of 100 %. The gene was found to be in all strains of and was highly conserved. was detected in 3.9 % (11/280, 95 % CI 2.1–6.9) of all male specimens, in 7.7 % (10/130, 95 % CI 4.1–13.7) of patients with non-gonococcal urethritis (NGU) and in 0.7 % (1/150, 95 % CI <0.01–4.1) of patients without urethritis. The presence of was significantly associated with NGU ( ≤0.01; 95 % Cl 0.88–0.98) and non-chlamydial-non-gonococcal urethritis (=0.0005; 95 % Cl 0.84–0.97).

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Cp, crossing point threshold , IPC, internal processing control , NCNGU, non-chlamydial-non-gonococcal urethritis and NGU, non-gonococcal urethritis
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2009-07-01
2023-01-30
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References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410 [CrossRef]
     [Google Scholar]
  2. Baseman J. B., Dallo S. F., Tully J. G., Rose D. L. 1988; Isolation and characterization of Mycoplasma genitalium strains from the human respiratory tract. J Clin Microbiol 26:2266–2269
     [Google Scholar]
  3. Björnelius E., Lidbrink P., Jensen J. S. 2000; Mycoplasma genitalium in non-gonococcal urethritis: a study in Swedish male STD patients. Int J STD AIDS 11:292–296 [CrossRef]
     [Google Scholar]
  4. Blom N., Gammeltoft S., Brunak S. 1999; Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. J Mol Biol 294:1351–1362 [CrossRef]
     [Google Scholar]
  5. Burgos R., Pich O. Q., Querol E., Piñol J. 2007; Functional analysis of the Mycoplasma genitalium MG312 protein reveals a specific requirement of the MG312 N-terminal domain for gliding motility. J Bacteriol 189:7014–7023 [CrossRef]
     [Google Scholar]
  6. Chenna R., Sugawara H., Koike T., Lopez R., Gibson T. J., Higgins D. G., Thompson J. D. 2003; Multiple sequence alignment with the clustal series of programs. Nucleic Acids Res 31:3497–3500 [CrossRef]
     [Google Scholar]
  7. Clausen H. F., Fedder J., Drasbek M., Nielsen P. K., Toft B., Ingerslev H. J., Birkelund S., Christiansen G. 2001; Serological investigation of Mycoplasma genitalium in infertile women. Hum Reprod 16:1866–1874 [CrossRef]
     [Google Scholar]
  8. Deguchi T., Yoshida T., Yokoi S., Ito M., Tamaki M., Ishiko H., Maeda S.-I. 2002; Longitudinal quantitative detection by real-time PCR of Mycoplasma genitalium in first-pass urine of men with recurrent nongonococcal urethritis. J Clin Microbiol 40:3854–3856 [CrossRef]
     [Google Scholar]
  9. Dhandayuthapani S., Rasmussen W. G., Baseman J. B. 2002; Stability of cytadherence-related proteins P140/P110 in Mycoplasma genitalium requires MG218 and unidentified factors. Arch Med Res 33:1–5 [CrossRef]
     [Google Scholar]
  10. Edberg A., Jurstrand M., Johansson E., Wikander E., Höög A., Ahlqvist T., Falk L., Jensen J. S., Fredlund H. 2008; A comparative study of three different PCR assays for detection of Mycoplasma genitalium in urogenital specimens from men and women. J Med Microbiol 57:304–309 [CrossRef]
     [Google Scholar]
  11. Gnad F., Ren S., Cox J., Olsen J. V., Macek B., Oroshi M., Mann M. 2007; PHOSIDA (phosphorylation site database): management, structural and evolutionary investigation, and prediction of phosphosites. Genome Biol 8:R250 [CrossRef]
     [Google Scholar]
  12. Hjorth S. V., Björnelius E., Lidbrink P., Falk L., Dohn B., Berthelsen L., Ma L., Martin D. H., Jensen J. S. 2006; Sequence-based typing of Mycoplasma genitalium reveals sexual transmission. J Clin Microbiol 44:2078–2083 [CrossRef]
     [Google Scholar]
  13. Horner P. J., Gilroy C. B., Thomas B. J., Naidoo R. O. M., Taylor-Robinson D. 1993; Association of Mycoplasma genitalium with acute non-gonococcal urethritis. Lancet 342:582–585 [CrossRef]
     [Google Scholar]
  14. Jensen J. S. 2006; Mycoplasma genitalium infections. Diagnosis, clinical aspects, and pathogenesis. Dan Med Bull 53:1–27
     [Google Scholar]
  15. Jensen J. S., Björnelius E., Dohn B., Lidbrink P. 2004; Use of TaqMan 5′ nuclease real-time PCR for quantitative detection of Mycoplasma genitalium DNA in males with and without urethritis who were attendees at a sexually transmitted disease clinic. J Clin Microbiol 42:683–692 [CrossRef]
     [Google Scholar]
  16. Keane F. E., Thomas B. J., Gilroy C. B., Renton A., Taylor-Robinson D. 2000; The association of Chlamydia trachomatis and Mycoplasma genitalium with non-gonococcal urethritis: observations on heterosexual men and their female partners. Int J STD AIDS 11:435–459 [CrossRef]
     [Google Scholar]
  17. Lind K., Lindhardt B. O., Schütten H. J., Blom J., Christiansen C. 1984; Serological cross-reactions between Mycoplasma genitalium and Mycoplasma pneumoniae . J Clin Microbiol 20:1036–1043
     [Google Scholar]
  18. Longo M. C., Berninger M. S., Hartley J. L. 1990; Use of uracil DNA glycosylate to control carry-over in polymerase chain reaction. Gene 93:125–128 [CrossRef]
     [Google Scholar]
  19. Ma L., Taylor S., Jensen J. S., Myers L., Lillis R., Martin D. H. 2008; Short tandem repeat sequences in the Mycoplasma genitalium genome and their use in a multilocus genotyping system. BMC Microbiol 8:130 [CrossRef]
     [Google Scholar]
  20. Maeda S. I., Tamaki M., Kojima K., Yoshida T., Ishiko H., Yasuda M., Deguchi T. 2001; Association of Mycoplasma genitalium persistence in the urethra with recurrence of nongonococcal urethritis. Sex Transm Dis 28:472–476 [CrossRef]
     [Google Scholar]
  21. Moi H., Reinton N., Moghaddam A. 2009; Mycoplasma genitalium is associated with symptomatic and asymptomatic non-gonococcal urethritis in men. Sex Transm Infect 85:15–18 [CrossRef]
     [Google Scholar]
  22. Musatovova O., Dhandayuthapani S., Baseman J. B. 2003; Transcriptional starts for cytadherence-related operons of Mycoplasma genitalium . FEMS Microbiol Lett 229:73–81 [CrossRef]
     [Google Scholar]
  23. Pich O. Q., Burgos R., Ferrer-Navarro M., Querol E., Piñol J. 2008; Role of Mycoplasma genitalium MG218 and MG317 cytoskeletal proteins in terminal organelle organization, gliding motility and cytadherence. Microbiology 154:3188–3198 [CrossRef]
     [Google Scholar]
  24. Siew N., Azaria Y., Fischer D. 2004; The ORFanage: an ORFan database. Nucleic Acids Res 32:D281–D283 [CrossRef]
     [Google Scholar]
  25. Su H. C., Hutchison C. A. III, Giddings M. C. 2007; Mapping phosphoproteins in Mycoplasma genitalium and Mycoplasma pneumoniae . BMC Microbiol 7:63 [CrossRef]
     [Google Scholar]
  26. Taylor-Robinson D., Furr P. M., Hanna N. F. 1985; Microbiological and serological study of non-gonococcal urethritis with special reference to Mycoplasma genitalium . Genitourin Med 61:319–324
     [Google Scholar]
  27. Taylor-Robinson D., Gilroy C. B., Keane F. E. 2003; Detection of several Mycoplasma species at various anatomical sites of homosexual men. Eur J Clin Microbiol Infect Dis 22:291–293
     [Google Scholar]
  28. Tully J. G., Taylor-Robinson D., Cole R. M., Rose D. L. 1981; A newly discovered mycoplasma in the human urogenital tract. Lancet 1:1288–1291
     [Google Scholar]
  29. Tully J. G., Rose D. L., Baseman J. B., Dallo S. F., Lazzell A. L., Davis C. P. 1995; Mycoplasma pneumoniae and Mycoplasma genitalium mixture in synovial fluid isolate. J Clin Microbiol 33:1851–1855
     [Google Scholar]
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Real-time PCR detection of the mg219 gene of unknown function of Mycoplasma genitalium in men with and without non-gonococcal urethritis and their female partners in England
J. Med. Microbiol. 58, 895 (2009); https://doi.org/10.1099/jmm.0.009977-0
/content/journal/jmm/10.1099/jmm.0.009977-0
/content/journal/jmm/10.1099/jmm.0.009977-0
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