Sensitive and specific assay for the simultaneous detection of Mycoplasma genitalium and macrolide resistance-associated mutations

Collection Location Koleksi E-book & E-Journal Perpustakaan Pusat Unila
Edition Vol. 37, Issue. 10
Call Number
ISBN/ISSN 1435-4373
Author(s) Braam, Joyce F...[et al.]
Subject(s) Biomedicine
Classification NONE
Series Title
GMD E-Journal
Language English
Publisher Springer
Publishing Year 2018
Publishing Place Switzerland
Collation
Abstract/Notes Abstract
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 resistanceassociated
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.
Keywords Azithromycin resistance . qPCR . Mycoplasma genitalium . 23S rRNA . A2058T
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