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01.12.2019 | Research article | Ausgabe 1/2019 Open Access

BMC Infectious Diseases 1/2019

Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium

Zeitschrift:
BMC Infectious Diseases > Ausgabe 1/2019
Autoren:
Olivier Thellin, Benaïssa Elmoualij, Willy Zorzi, Jorgen S. Jensen, Renaud Close, Valerie Deregowski, Muriel Le Guern Fellous, Pascale Quatresooz
Wichtige Hinweise

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Abstract

Background

The worldwide expansion of macrolide-resistant Mycoplasma genitalium (MG) in cases of genital infections has led to an increased recurrence rate of these infections after first-line azithromycin treatment. By detecting the presence of azithromycin-resistant MG, the patient’s antibiotic treatment can be targeted and the spread of resistance prevented. With this aim in mind, macrolide-resistance detection kits are helpful tools for the physician.

Methods

Azithromycin resistance mutations in MG are targeted using a four-color multiplex real-time RT-PCR assay. Tested targets include plasmid DNA (as positive controls) as well as macrolide-sensitive and macrolide-resistant genomic DNA from characterized cell lines and clinical samples.

Results

The analytical data presented here were generated from plasmid DNA and genomic RNA/DNA and include adaptation to an internal control, specificity between targets, specificity vs non-MG species, limit of detection (LoD) and interference studies (co-infection and endogenous substances). The clinical data were based on the application of the assay to clinical samples characterized by sequencing.

Conclusions

A new NAAT (nucleic acid amplification test) prototype has been developed in collaboration with the Diagenode s.a. company, this prototype targets MG and azithromycin-resistance mutations in that pathogen.
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