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

[en] 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. © 2019 The Author(s).

Disciplines :

Immunology & infectious disease

Author, co-author :

Thellin, Olivier ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Histologie

Elmoualij, Benaïssa ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Histologie

Zorzi, Willy ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Histologie

Jensen, J. S.; Statens Serum Institut, Artillerivej 5, Copenhagen S, 2300, Denmark

Close, R.; Diagenode S.a., Rue du Bois Saint-Jean 3, Liège, 4102, Belgium

Deregowski, V.; Diagenode S.a., Rue du Bois Saint-Jean 3, Liège, 4102, Belgium

Le Guern Fellous, M.; Diagenode S.a., Rue du Bois Saint-Jean 3, Liège, 4102, Belgium

Quatresooz, Pascale ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Histologie

Language :

English

Title :

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

Publication date :

2019

Journal title :

BMC Infectious Diseases

eISSN :

1471-2334

Publisher :

BioMed Central Ltd.

Volume :

Issue :

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 07 August 2020

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Bibliography

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