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Comparison of gradient diffusion and molecular methods using Allplex™ NG&DR assay (Seegene®) for macrolide and fluoroquinolone screening resistance in Neisseria gonorrhoeae

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Abstract

Antimicrobial resistance in Neisseria gonorrhoeae (NG) is increasing worldwide. Second-line treatments with macrolides or fluoroquinolones are an option for NG infections in some cases following the STI guideline recommendations. In our study, we compared the gradient diffusion test using EUCAST 2024 breakpoints with a new molecular method using the Allplex™ NG&DR assay (Seegene®) including A2059G/C2611 mutations (23S rRNA) associated with high/moderate-level macrolide resistance and S91F mutation (gyrA) relationship with fluoroquinolone resistance in NG isolates (n = 100). We calculated the sensitivity, specificity, and correlation of the molecular test for fluoroquinolone using the gradient diffusion as the reference method. In twenty-three strains was not detected any mutation associated with macrolides or fluoroquinolone resistance. No A2059G/C2611T mutations were detected, and the S91F mutations were detected in 77 out of the 100 isolates screened. Twenty-three NG isolates were reported to be resistant to azithromycin (ECOFF: >1 mg/L), and 78 NG isolates were resistant to ciprofloxacin (MIC: >0.06 mg/L). The molecular method showed a sensitivity of 96.1% and, a specificity of 90.9% for fluoroquinolone susceptibility, but the statistical analysis between the molecular test and gradient diffusion test was not statistically significant for fluoroquinolone resistance (p = 1). Statistical analysis was not performed for macrolides because of the absence of positive RT-PCR results. According to our data, Allplex™ assay cannot replace the gradient diffusion test for macrolide resistance. However, the assay could be used to test fluoroquinolone resistance in NG isolates as a replacement for phenotypic methods.

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Acknowledgements

To my mentor and friend, Inmaculada Quiles Melero, for trusting me during these years and supporting me in my personal and professional life.

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Not funding.

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Author notes

  1. Alfredo Maldonado-Barrueco and Claudia Sanz-González contributed equally to this work.

Authors and Affiliations

  1. Clinical Microbiology Department, Hospital Universitario La Paz, 261, Madrid, 28046, Spain

    Alfredo Maldonado-Barrueco, Claudia Sanz-González, Iker Falces-Romero, Paloma García-Clemente, Juana Cacho-Calvo & Inmaculada Quiles-Melero

  2. CIBERINFEC, Instituto de Salud Carlos III, Madrid, Spain

    Iker Falces-Romero

Authors
  1. Alfredo Maldonado-Barrueco
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  2. Claudia Sanz-González
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  3. Iker Falces-Romero
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  4. Paloma García-Clemente
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  5. Juana Cacho-Calvo
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  6. Inmaculada Quiles-Melero
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Contributions

AMB, CSG: design, analysis of data, and writing of the manuscript. IFR: serological data, review of the manuscript. PGC: design, review of the manuscript. JCC: review of the manuscript. IQM: design, writing of the manuscript, and review of the manuscript.

Corresponding author

Correspondence to Alfredo Maldonado-Barrueco.

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Ethical review and approval were not required for the study on human participants in accordance with the local legislation and institutional requirements. The data used are from clinical samples that were necessary for the clinical STI diagnosis of the patients. The detection of resistance associated with macrolide and fluoroquinolone in Neisseria gonorrhoeae was not incorporated into the medical clinical reports of the patients.

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Maldonado-Barrueco, A., Sanz-González, C., Falces-Romero, I. et al. Comparison of gradient diffusion and molecular methods using Allplex™ NG&DR assay (Seegene®) for macrolide and fluoroquinolone screening resistance in Neisseria gonorrhoeae. Eur J Clin Microbiol Infect Dis 43, 1009–1012 (2024). https://doi.org/10.1007/s10096-024-04786-y

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