Sensitive and specific assay for the simultaneous detection of Mycoplasma genitalium and macrolide resistance-associated mutations
Autor: | Tim T Severs, Yevgeniy S. Belousov, Walt Mahoney, Joyce F Braam, Sebastian van Marm, Johannes G Kusters |
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Přispěvatelé: | Integrative Neurophysiology |
Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Microbiology (medical) Azithromycin resistance medicine.medical_specialty Routine testing 030106 microbiology Mycoplasma genitalium Azithromycin 03 medical and health sciences 0302 clinical medicine Medical microbiology SDG 3 - Good Health and Well-being 23S ribosomal RNA medicine 030212 general & internal medicine Gene A2058T biology Macrolide resistant 23S rRNA General Medicine biology.organism_classification Molecular biology qPCR Infectious Diseases Macrolide resistance medicine.drug |
Zdroj: | European Journal of Clinical Microbiology and Infectious Diseases, 37(11), 2137-2144. Springer Verlag Braam, J F, van Marm, S, Severs, T T, Belousov, Y, Mahoney, W & Kusters, J G 2018, ' Sensitive and specific assay for the simultaneous detection of Mycoplasma genitalium and macrolide resistance-associated mutations ', European Journal of Clinical Microbiology and Infectious Diseases, vol. 37, no. 11, pp. 2137-2144 . https://doi.org/10.1007/s10096-018-3350-3 |
ISSN: | 0934-9723 |
Popis: | Patients infected by Mycoplasma genitalium are often treated empirically with the macrolide azithromycin. Macrolide resistance is becoming quite common; empirical treatment is compromised. Sequencing was initially used to detected azithromycin resistance-associated mutations. As this was laborious, qPCRs have been developed for their detection. In the present study, we describe a fast, sensitive, and specific qPCR assay that enables routine testing of M. genitalium and macrolide resistance-associated mutations in a single assay. M. genitalium positive clinical samples were used to compare (i) the commonly used MgPa assay for the detection of M. genitalium infections (MgPa qPCR), (ii) a combined 23S rRNA gene PCR/sequencing assay (Mg23S qPCR/Sequencing) to identify macrolide resistance-associated mutations, and (iii) our newly developed probe-based melt curve qPCR for simultaneous detection of M. genitalium and macrolide resistance-associated mutations (Macrolide-R/MG ELITe MGB Kit, Elitech Bothel USA in short Mg MacrolideR qPCR). Specificity of the qPCR was tested using urogenital samples that were tested positive for a range of other micro-organisms. M. genitalium was detected in 196/236 (83.1%) samples by the MgPa qPCR, versus 172/236 (72.9%) by the combined Mg23S qPCR/Sequencing, and 202/236 (85.6%) by the Mg MacrolideR qPCR. The Mg MacrolideR qPCR showed high concordance to the Mg23S qPCR/Sequencing assay (201 vs 202 could be genotyped, respectively) for the detection of the macrolide resistant mutations. None of the other urogenital pathogens were tested positive in the Mg MacrolideR qPCR, indicating specificity. The Mg MacrolideR qPCR is fast, sensitive, specific, and can easily be implemented in the routine diagnostics. |
Databáze: | OpenAIRE |
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