Abstract
Urogenital tract infections can be caused by a number of pathogens, some of which, like the obligate intracellular Chlamydia trachomatis, are difficult to culture, or the cell wall-less mollicutes, like M. hominis or Ureaplasma spp. Real-time PCR (qPCR) has become an important diagnostic tool as it enables not only the species-specific detection of the organism but also the quantification essential to define the etiological relevance of a facultative pathogenic bacterium. We developed a set of TaqMan qPCRs for the detection of the species M. genitalium and M. hominis (Mh/Mg-duplex qPCR), U. parvum and U. urealyticum (Uu/Up duplex-PCR), and C. trachomatis (CT-qPCR), and for typing of lymphogranuloma venereum-associated L-serovars of C. trachomatis (LGV-qPCR) as well as a sub-typing of L1, L2, and L3. In addition, the human gap-gene was amplified as quality control of the specimen, and a cryptic plasmid co-amplified in CT-qPCR as an inhibition control. The present protocol focuses on the step-by-step description for the establishment of these TaqMan multiplex qPCRs.
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Acknowledgements
We would like to especially thank Servas Morré and Eberhard Straube for kindly providing the Chlamydia strains.
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Möbius, N., Brenneisen, W., Schaeffer, A., Henrich, B. (2012). Protocol for the Rapid Detection of the Urogenital Tract Mollicutes and Chlamydia with Concomitant LGV-(sub)typing. In: MacKenzie, C., Henrich, B. (eds) Diagnosis of Sexually Transmitted Diseases. Methods in Molecular Biology, vol 903. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-937-2_15
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DOI: https://doi.org/10.1007/978-1-61779-937-2_15
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Publisher Name: Humana Press, Totowa, NJ
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