Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium
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 anti...
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| Format: | Article |
| Language: | English |
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BMC
2019-09-01
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| Series: | BMC Infectious Diseases |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12879-019-4424-2 |
| _version_ | 1818273978800668672 |
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| author | Olivier Thellin Benaïssa Elmoualij Willy Zorzi Jorgen S. Jensen Renaud Close Valerie Deregowski Muriel Le Guern Fellous Pascale Quatresooz |
| author_facet | Olivier Thellin Benaïssa Elmoualij Willy Zorzi Jorgen S. Jensen Renaud Close Valerie Deregowski Muriel Le Guern Fellous Pascale Quatresooz |
| author_sort | Olivier Thellin |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-12T22:06:33Z |
| format | Article |
| id | doaj.art-23646d821d514a239ceadb2310a337c3 |
| institution | Directory Open Access Journal |
| issn | 1471-2334 |
| language | English |
| last_indexed | 2024-12-12T22:06:33Z |
| publishDate | 2019-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Infectious Diseases |
| spelling | doaj.art-23646d821d514a239ceadb2310a337c32022-12-22T00:10:21ZengBMCBMC Infectious Diseases1471-23342019-09-011911810.1186/s12879-019-4424-2Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitaliumOlivier Thellin0Benaïssa Elmoualij1Willy Zorzi2Jorgen S. Jensen3Renaud Close4Valerie Deregowski5Muriel Le Guern Fellous6Pascale Quatresooz7Department of Human Histology-CRPP, University of LiègeDepartment of Human Histology-CRPP, University of LiègeDepartment of Human Histology-CRPP, University of LiègeStatens Serum InstitutDiagenode s.a.Diagenode s.a.Diagenode s.a.Department of Human Histology-CRPP, University of LiègeAbstract 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.http://link.springer.com/article/10.1186/s12879-019-4424-2Real-time PCRMycoplasma genitaliumAzithromycin resistanceDiagnostic |
| spellingShingle | Olivier Thellin Benaïssa Elmoualij Willy Zorzi Jorgen S. Jensen Renaud Close Valerie Deregowski Muriel Le Guern Fellous Pascale Quatresooz Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium BMC Infectious Diseases Real-time PCR Mycoplasma genitalium Azithromycin resistance Diagnostic |
| title | Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium |
| title_full | Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium |
| title_fullStr | Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium |
| title_full_unstemmed | Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium |
| title_short | Four-color multiplex real-time PCR assay prototype targeting azithromycin resistance mutations in Mycoplasma genitalium |
| title_sort | four color multiplex real time pcr assay prototype targeting azithromycin resistance mutations in mycoplasma genitalium |
| topic | Real-time PCR Mycoplasma genitalium Azithromycin resistance Diagnostic |
| url | http://link.springer.com/article/10.1186/s12879-019-4424-2 |
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