Background Sexually transmitted infections are caused by a relatively well defined group of pathogens. Their individual detection using cultural and molecular techniques is time-consuming and costly. Multiplex real-time PCR is a rapid and more cost-effective alternative and allows the detection of multiple infections.
Methods We have validated the Anyplex IITM STI-7, a semiquantitative, highly multiplexed real-time PCR kit (Seegene), using a selection of specimens positive by routine methods (culture, PCR, cytology) for at least one of the 7 different targets. Specimens were assumed to be negative for those parameters not previously tested. DNA was isolated using the easyMAG® (bioMérieux) followed by melting curve analysis-based PCR on a CFX96TM thermocycler (BioRad) and automatic data interpretation with the Seegene Viewer software. Discrepant results were resolved with independent molecular tests.
Results Resolved results showed 100% sensitivity and specificity for Chlamydia trachomatis (17 positive/73 negative specimens), Neisseria gonorrhoeae (13/71), Trichomonas vaginalis (6/84), Mycoplasma genitalium (18/72) and Mycoplasma hominis (30/60). For Ureaplasma species (57/30) 100% sensitivity and 93.3% specificity were observed. The STI-7 test (the only test capable of separating the two major species) revealed that among the 57 Ureaplasma-positive specimens 7 (12.3%) were positive for U. urealyticum, 39 (68.4%) for U. parvum and 11 (19.3%) for both species. Often, 2 or more targets were detected, e.g. of the 17 C. trachomatis-positive specimens 8 were positive for 1 and 7 for 2 additional organisms. Ureaplasmas were the most prevalent species being present in about 2/3 of the specimens.
Conclusion We conclude that the STI-7 multiplex PCR is a rapid and reliable test for the simultaneous detection of the most important sexually transmitted pathogens providing an efficient means for a more thorough evaluation of the clinical significance of the various organisms.
- multiple infections
- multiplex detection
- Real-time PCR
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.