A real-time PCR assay for detecting a penA mutation associated with ceftriaxone resistance in Neisseria gonorrhoeae
| Autor: | Ken Shimuta, Shu-ichi Nakayama, Makoto Ohnishi, Mitsuru Yasuda, Takashi Deguchi, Gene Igawa |
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| Rok vydání: | 2019 |
| Předmět: |
DNA
Bacterial 0301 basic medicine Microbiology (medical) 030106 microbiology Immunology Microbial Sensitivity Tests Biology Real-Time Polymerase Chain Reaction medicine.disease_cause Microbiology Gonorrhea 03 medical and health sciences 0302 clinical medicine Antibiotic resistance Drug Resistance Bacterial medicine Humans Penicillin-Binding Proteins Immunology and Allergy 030212 general & internal medicine Gene Alleles Mutation Ceftriaxone Sequence Analysis DNA biology.organism_classification Neisseria gonorrhoeae Anti-Bacterial Agents genomic DNA Real-time polymerase chain reaction Genes Bacterial Neisseria Primer (molecular biology) Sequence Alignment |
| Zdroj: | Journal of Global Antimicrobial Resistance. 19:46-49 |
| ISSN: | 2213-7165 |
| Popis: | Objectives Ceftriaxone (CRO) resistance is spreading worldwide, and hindering the effective treatment of gonococcal infections. This study developed a detection system for the genomic DNA of CRO-resistant Neisseria gonorrhoeae (N. gonorrhoeae) strains, in order to improve the surveillance of antimicrobial resistance. Methods A real-time PCR assay targeting the penA gene of recently isolated CRO-resistant N. gonorrhoeae strains was designed. Primer and probe sequence information was obtained from sequence comparisons between penA of Neisseria spp. and penA of CRO-resistant N. gonorrhoeae strains. Results Using this assay, a positive reaction was observed using the genomic DNA of three strains (GU140106, FC428, and A8806). The assay was evaluated using genomic DNA of 204 N. gonorrhoeae and 95 Neisseria spp. isolates with known minimum inhibitory concentrations of CRO. Following PCR assays for these strains, three FC428-related strains were positively identified, which possessed penA-60.001, whereas the remaining 201 N. gonorrhoeae strains and 95 Neisseria spp. strains were negative. Conclusions A real-time PCR-based assay was designed to detect the genomic DNA of strains harbouring mosaic penA-59.001 (GU140106), penA-60.001 (FC428), and penA-64.001 (A8806) alleles and to discriminate them from N. gonorrhoeae and Neisseria spp. strains harbouring other genes. |
| Databáze: | OpenAIRE |
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