Real-time PCR detection of Neisseria gonorrhoeae susceptibility to penicillin

Buckley, Cameron, Trembizki, Ella, Donovan, Basil, Chen, Marcus, Freeman, Kevin, Guy, Rebecca, Lahra, Monica M., Kundu, Ratan L., Regan, David G., Smith, Helen V. and Whiley, David M. (2016) Real-time PCR detection of Neisseria gonorrhoeae susceptibility to penicillin. Journal of Antimicrobial Chemotherapy, . doi:10.1093/jac/dkw291


Author Buckley, Cameron
Trembizki, Ella
Donovan, Basil
Chen, Marcus
Freeman, Kevin
Guy, Rebecca
Lahra, Monica M.
Kundu, Ratan L.
Regan, David G.
Smith, Helen V.
Whiley, David M.
Title Real-time PCR detection of Neisseria gonorrhoeae susceptibility to penicillin
Formatted title
Real-time PCR detection of Neisseria gonorrhoeae susceptibility to penicillin
Journal name Journal of Antimicrobial Chemotherapy   Check publisher's open access policy
ISSN 1460-2091
0305-7453
Publication date 2016-07-25
Year available 2016
Sub-type Article (original research)
DOI 10.1093/jac/dkw291
Open Access Status Not Open Access
Total pages 6
Place of publication Oxford, United Kingdom
Publisher Oxford University Press
Collection year 2017
Language eng
Formatted abstract
Objectives: The objective of this study was to develop a real-time PCR assay targeting the gonococcal porB gene (PorB-PCR) for predicting susceptibility of Neisseria gonorrhoeae to penicillin. This complements a previously described PCR assay for detecting penicillinase-producing N. gonorrhoeae (PPNG) developed by our laboratory (PPNG-PCR).

Methods: The PorB-PCR assay was designed using six probes to characterize various combinations of amino acids at positions 101 and 102 of the PorB1b class protein, including the WT G101/A102 and mutant G101K/A102D, G101K/A102N and G101K/A102G sequences, as well as the PorB1a sequence. The ability of these sequences to predict penicillin susceptibility was initially assessed using 2307 N. gonorrhoeae isolates from throughout Australia for which phenotypic susceptibility data were available. The assay was then applied to N. gonorrhoeae-positive clinical specimens (n = 70). Specificity was assessed by testing commensal Neisseria strains (n = 75) and N. gonorrhoeae-negative clinical specimens (n = 171).

Results: Testing of the 2307 N. gonorrhoeae isolates using PorB-PCR to detect G101/A102 and PorB1a sequences identified a total of 78.4% (61.2% and 17.2%, respectively) of penicillin-susceptible isolates with specificities of 97.4% and 99.3% and positive predictive values of 98.8% and 98.9%, where PPNG strains were simultaneously identified and excluded. Similar performance data were obtained when the PorB-PCR assay was applied to the N. gonorrhoeae-positive clinical specimens. No false-positive results were observed for the N. gonorrhoeae-negative samples and no cross-reactions were observed with the non-gonococcal species.

Conclusions: When used in parallel with the previously described PPNG-PCR, the PorB-PCR approach has the potential to facilitate individualized treatment of gonorrhoea using penicillin.
Keyword Neisseria gonorrhoeae
N. gonorrhoeae
Penicillin
Penicillinase-producing N. gonorrhoeae (PPNG)
Individualized treatment
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: UQ Centre for Clinical Research Publications
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Created: Mon, 01 Aug 2016, 10:09:48 EST by Ella Trembizki on behalf of UQ Centre for Clinical Research