A real-time PCR assay for detecting a penA mutation associated with ceftriaxone resistance in Neisseria gonorrhoeae. (December 2019)

• Record Type: Journal Article
• Authors: Shimuta, Ken; Igawa, Gene; Yasuda, Mitsuru; Deguchi, Takashi; Nakayama, Shu-ichi; Ohnishi, Makoto
• Is Part Of: Journal of global antimicrobial resistance. Volume 19(2019)
• Page Start: 46
• Page End: 49
• Publication Date: 2019-12
• Subjects: Ceftriaxone (CRO) -- Neisseria gonorrhoeae -- penA; Drug resistance -- Periodicals; Drug resistance -- Periodicals; Drug resistance; Periodicals; 616.9041
• DOI: 10.1016/j.jgar.2019.02.011
• Languages: English
• ISSNs: 2213-7165

Abstract

Highlights: A PCR assay was developed for targeting a penA mutation associated with ceftriaxone resistance. The genomic DNA of Neisseria gonorrhoeae strains harboring mosaic penA -59.001, penA -60.001, and penA -64.001 alleles was reliably detected. Ninety-five and 201 Neisseria spp. and N. gonorrhoeae isolates, respectively, were negative. This assay may be useful for molecular surveillance using culture-negative specimens. The proposed assay can make surveillance of antimicrobial resistance in N. gonorrhoeae more effective. Abstract: 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.