Hydrogel Droplet Microarray for Genotyping Antimicrobial Resistance Determinants in <i>Neisseria gonorrhoeae</i> Isolates
A multiplex assay based on a low-density hydrogel microarray was developed to identify genomic substitutions in <i>N. gonorrhoeae</i> that determine resistance to the currently recommended treatment agents ceftriaxone and azithromycin and the previously used drugs penicillin, tetracyclin...
Guardado en:
| Autores principales: | , , , , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/afb8f554e1b64dafaf39ff1d0981580b |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| Sumario: | A multiplex assay based on a low-density hydrogel microarray was developed to identify genomic substitutions in <i>N. gonorrhoeae</i> that determine resistance to the currently recommended treatment agents ceftriaxone and azithromycin and the previously used drugs penicillin, tetracycline, and ciprofloxacin. The microarray identifies 74 drug resistance determinants in the <i>N. gonorrhoeae</i> <i>penA</i>, <i>ponA</i>, <i>porB</i>, <i>gyrA</i>, <i>parC</i>, <i>rpsJ</i>, <i>mtrR</i>, <i>bla</i><sub>TEM</sub>, <i>tetM</i>, and 23S rRNA genes. The hydrogel elements were formed by automated dispensing of nanoliter-volume droplets followed by UV-induced copolymerization of NH<sub>2</sub>-containing oligonucleotides with gel-forming monomers. Polybutylene terephthalate plates without special modifications were used as microarray substrates. Sequences and concentrations of immobilized oligonucleotides, gel composition, and hybridization conditions were carefully selected, and the median discrimination ratio ranged from 2.8 to 29.4, allowing unambiguous identification of single-nucleotide substitutions. The mutation identification results in a control sample of 180 <i>N. gonorrhoeae</i> isolates were completely consistent with the Sanger sequencing results. In total, 648 clinical <i>N. gonorrhoeae</i> isolates obtained in Russia during the last 5 years were analyzed and genotyped using these microarrays. The results allowed us to draw conclusions about the present situation with antimicrobial susceptibility of <i>N. gonorrhoeae</i> in Russia and demonstrated the possibility of using hydrogel microarrays to control the spread of antibiotic resistance. |
|---|