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- Title
Enhanced gonococcal antimicrobial surveillance in the era of ceftriaxone resistance: a real-time PCR assay for direct detection of the Neisseria gonorrhoeae H041 strain.
- Authors
Goire, Namraj; Ohnishi, Makoto; Limnios, Athena E.; Lahra, Monica M.; Lambert, Stephen B.; Nimmo, Graeme R.; Nissen, Michael D.; Sloots, Theo P.; Whiley, David M.
- Abstract
Objectives Recent emergence of the extensively drug-resistant Neisseria gonorrhoeae H041 strain in Japan raises concerns that gonorrhoea may soon become untreatable and emphasizes the need for enhanced surveillance. In this study we developed a real-time PCR assay for direct detection of the H041 strain. Methods Two real-time PCR assays for detection of the penA gene of the H041 strain, H041-PCR1 and H041-PCR2, were developed and evaluated in parallel. Assay performance was assessed using a panel of pathogenic and commensal Neisseria species (n = 167 strains) including the N. gonorrhoeae H041 strain and clinical specimens (n = 252) submitted for sexual health screening. The detection limits of the assays were compared with a standard N. gonorrhoeae real-time PCR method. Results Both the H041-PCR1 and H041-PCR2 assays correctly detected the N. gonorrhoeae H041 strain and provided negative results for all other N. gonorrhoeae strains. However, only the H041-PCR2 assay proved to be specific when applied to the non-gonococcal Neisseria species and clinical samples. False-positive results in the H041-PCR1 included cross-reactions with two Neisseria subflava isolates and eight clinical specimens. DNA sequencing of these N. subflava strains revealed the presence of the penicillin-binding protein 2 Ala328Thr alteration previously only observed in the N. gonorrhoeae H041 strain. Conclusions The H041-PCR2 assay is suitable for direct detection of the N. gonorrhoeae H041 ceftriaxone-resistant strain in cultured and non-cultured samples.
- Subjects
JAPAN; DRUG resistance in microorganisms; NEISSERIA gonorrhoeae; POLYMERASE chain reaction; MICROBIOLOGICAL assay
- Publication
Journal of Antimicrobial Chemotherapy (JAC), 2012, Vol 67, Issue 4, p902
- ISSN
0305-7453
- Publication type
Article
- DOI
10.1093/jac/dkr549