Multiplex real-time PCR assay targeting macrolide resistance mutations in Mycoplasma genitalium

2016 ◽  
Author(s):  
Olivier Thellin
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Mycoplasma Genitalium ◽  
Macrolide Resistance ◽  
Resistance Mutations ◽  
Pcr Assay

scholarly journals Evaluation of a real-time PCR assay for detection of Mycoplasma genitalium and macrolide resistance-mediating mutations from clinical specimens

2018 ◽  
Vol 91 (2) ◽  
pp. 123-125 ◽  
Author(s):  
Li Xiao ◽  
Ken B. Waites ◽  
Hong Wang ◽  
Barbara Van Der Pol ◽  
Amy E. Ratliff ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Mycoplasma Genitalium ◽  
Macrolide Resistance ◽  
Pcr Assay ◽  
Clinical Specimens

scholarly journals Performance of Three Commercial Molecular Diagnostic Assays for the Simultaneous Detection of Mycoplasma genitalium and Macrolide Resistance

2021 ◽  
Author(s):  
Chloé Le Roy ◽  
Cécile Bébéar ◽  
Sabine Pereyre
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Mycoplasma Genitalium ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Molecular Diagnostic ◽  
Handling Characteristics ◽  
Significant Difference ◽  
Commercial Kit ◽  
Resistance Detection

The increasing frequency of macrolide resistance is an emerging issue in the treatment of Mycoplasma genitalium infection. Because evaluation of new commercial kit detecting M. genitalium and macrolide resistance is needed, we evaluated the performance and handling characteristics of the Allplex MG & AziR (Seegene), the Macrolide-R/MG ELITe MGB (ELITechGroup), and the ResistancePlus MG FleXible kits (SpeeDx-Cepheid) in comparison with an in-house real-time PCR and 23S rRNA gene sequencing used as reference. A total of 239 urogenital specimens (135 M. genitalium-positive and 104 M. genitalium-negative specimens) collected between April and December 2019 at the French National Reference Center for bacterial Sexually Transmitted Infections were assessed. The overall agreement for M. genitalium detection of the three commercial kits compared with the in-house real-time PCR was 94.6–97.6%, and there was no significant difference. A total of 97 specimens were found M. genitalium-positive with the three kits and were used to assess macrolide resistance detection. The clinical sensitivity for resistance detection was 74.5% (95% confidence interval 61.7–84.2%), 96.2% (87.2–99.0%), and 92.8% (82.7–97.1%) for the Allplex MG & AziR, Macrolide-R/MG ELITe MGB, and ResistancePlusMG FleXible kits, respectively. The sensitivity of the Macrolide-R/MG ELITe MGB kit was significantly higher than that of the Allplex MG & AziR kit. The clinical specificity for resistance detection of the three kits was 97.4–97.6%. The random-access possibility, input sample volume, and DNA extract availability for detecting resistance to other antibiotics may also influence the selection of a commercial kit by diagnostic laboratories.

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

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Olivier Thellin ◽  
Benaïssa Elmoualij ◽  
Willy Zorzi ◽  
Jorgen S. Jensen ◽  
Renaud Close ◽  
...  
Keyword(s):  
Real Time ◽  
Internal Control ◽  
Plasmid Dna ◽  
Limit Of Detection ◽  
Analytical Data ◽  
Mycoplasma Genitalium ◽  
Clinical Samples ◽  
Resistance Mutations ◽  
Pcr Assay ◽  
Azithromycin Resistance

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.

Evaluation of the new AmpliSens multiplex real-time PCR assay for simultaneous detection ofNeisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, andTrichomonas vaginalis

Apmis ◽  
2015 ◽  
Vol 123 (10) ◽  
pp. 879-886 ◽  
Author(s):  
Tatiana Rumyantseva ◽  
Daniel Golparian ◽  
Christian S. Nilsson ◽  
Emma Johansson ◽  
My Falk ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Mycoplasma Genitalium ◽  
Pcr Assay ◽  
Ofneisseria Gonorrhoeae
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