1887

Abstract

The aim of this study was to determine whether a patient's endocervical swab specimen can be transported in first void urine (FVU) as combined specimens for the detection of by real-time PCR. The study also compared two different DNA extraction methods for observation of possible PCR inhibition. Three specimens, one endocervical swab specimen transported in 2-SP medium, one endocervical swab specimen transported in FVU and a FVU specimen, were collected from 329 women. All sample types underwent manual DNA extraction whereas in the DNA extraction study, 329 endocervical swab specimens transported in FVU were subjected to both manual Chelex and automated BioRobot M48 DNA extraction. A total of 100 endocervical swab specimens transported in FVU from patients PCR-negative for in the study were used in the PCR inhibition analysis. was detected in 25/329 (7.6 %) women. The endocervical swab specimens transported in 2-SP medium and transported in FVU were positive for in 17/25 (68 %) and 24/25 (96 %) women, respectively. The FVU specimens alone were positive for in 22/25 (88 %) women. In the DNA extraction study, DNA was detected in 24/329 (7.3 %) and 28/329 (8.5 %) of endocervical swab specimens transported in FVU subjected to manual Chelex extraction and automated BioRobot M48 extraction, respectively. Partial PCR inhibition was detected in 6 % of samples subjected to manual Chelex extraction whereas no inhibition was detected with the automated BioRobot M48 extraction. Thus endocervical swab specimens transported in FVU demonstrate higher sensitivity than FVU specimens only and have considerably increased sensitivity compared with endocervical swab specimens transported in 2-SP medium for detection of DNA. Moreover, automated BioRobot M48 extraction was shown to be superior to a crude manual Chelex extraction, leaving no PCR inhibition and giving a slightly higher DNA yield and/or better sensitivity.

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2009-01-01
2024-03-28
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References

  1. Airell Å., Ottosson L., Bygdeman S. M., Carlberg H., Lidbrink P., Rudén A.-K., Elfgren K. 2000; Chlamydia trachomatis PCR (Cobas Amplicor) in women: endocervical specimen transported in a specimen of urine versus endocervical and urethral specimens in 2-SP medium versus urine specimen only. Int J STD AIDS 11:651–658 [CrossRef]
    [Google Scholar]
  2. Cohen C. R., Manhart L. E., Bukusi E. A., Astete S., Brunham R. C., Holmes K. K., Sinei S. K., Bwayo J. J., Totten P. A. 2002; Association between Mycoplasma genitalium and acute endometritis. Lancet 359:765–766 [CrossRef]
    [Google Scholar]
  3. Deguchi T., Maeda S. 2002; Mycoplasma genitalium : another important pathogen of nongonococcal urethritis. J Urol 167:1210–1217 [CrossRef]
    [Google Scholar]
  4. 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]
  5. Falk L., Fredlund H., Jensen J. S. 2004; Symptomatic urethritis is more prevalent in men infected with Mycoplasma genitalium than with Chlamydia trachomatis . Sex Transm Infect 80:289–293 [CrossRef]
    [Google Scholar]
  6. Falk L., Fredlund H., Jensen J. S. 2005; Signs and symptoms of urethritis and cervicitis among women with or without Mycoplasma genitalium or Chlamydia trachomatis infection. Sex Transm Infect 81:73–78 [CrossRef]
    [Google Scholar]
  7. Jensen J. S. 2004; Mycoplasma genitalium : the aetiological agent of urethritis and other sexually transmitted diseases. J Eur Acad Dermatol Venereol 18:1–11
    [Google Scholar]
  8. Jensen J. S., Björnelius E., Dohn B., Lidbrink P. 2004a; Comparison of first void urine and urogenital swab specimens for detection of Mycoplasma genitalium and Chlamydia trachomatis by polymerase chain reaction in patients attending a sexually transmitted disease clinic. Sex Transm Dis 31:499–507 [CrossRef]
    [Google Scholar]
  9. Jensen J. S., Björnelius E., Dohn B., Lidbrink P. 2004b; 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]
  10. Jurstrand M., Jensen J. S., Fredlund H., Falk L., Mölling P. 2005; Detection of Mycoplasma genitalium in urogenital specimens by real-time PCR and by conventional PCR assay. J Med Microbiol 54:23–29 [CrossRef]
    [Google Scholar]
  11. Taylor-Robinson D. 2002; Mycoplasma genitalium – an up-date. Int J STD AIDS 13:145–151 [CrossRef]
    [Google Scholar]
  12. 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]
  13. Uusküla A., Kohl P. K. 2002; Genital mycoplasmas, including Mycoplasma genitalium , as sexually transmitted agents. Int J STD AIDS 13:79–85 [CrossRef]
    [Google Scholar]
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