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  • Enhanced gonococcal antimicrobial surveillance in the era of ceftriaxone resistance: a real-time PCR assay for direct detection of the Neisseria gonorrhoeae H041 strain

    Author(s)
    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
    Griffith University Author(s)
    Nimmo, Graeme R.
    Year published
    2012
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    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 ...
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    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.
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    Journal Title
    Journal of Antimicrobial Chemotherapy
    Volume
    67
    Issue
    4
    DOI
    https://doi.org/10.1093/jac/dkr549
    Subject
    Microbiology
    Medical microbiology
    Pharmacology and pharmaceutical sciences
    Publication URI
    http://hdl.handle.net/10072/45629
    Collection
    • Journal articles

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