Please use this identifier to cite or link to this item: http://docs.prosentient.com.au/prosentientjspui/handle/1/10498
Title: Real-time PCR detection of Neisseria gonorrhoeae susceptibility to penicillin.
Authors: 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
Affiliation: The University of Queensland, UQ Centre for Clinical Research (UQCCR), Herston, Queensland 4029, Australia..
The University of Queensland, UQ Centre for Clinical Research (UQCCR), Herston, Queensland 4029, Australia e.trembizki@uq.edu.au..
Kirby Institute, UNSW Australia, Sydney, New South Wales 2052, Australia..
Melbourne Sexual Health Centre, Alfred Health, Carlton, Victoria 3053, Australia.. Central Clinical School, Monash University, Melbourne, Victoria 3181, Australia..
Microbiology Laboratory, Pathology Department, Royal Darwin Hospital, Darwin, Northern Territory, Australia..
Kirby Institute, UNSW Australia, Sydney, New South Wales 2052, Australia..
WHO Collaborating Centre for STD, Microbiology Department, South Eastern Area Laboratory Services, Prince of Wales Hospital, Sydney, New South Wales 2031, Australia..
WHO Collaborating Centre for STD, Microbiology Department, South Eastern Area Laboratory Services, Prince of Wales Hospital, Sydney, New South Wales 2031, Australia..
Kirby Institute, UNSW Australia, Sydney, New South Wales 2052, Australia..
Public Health Microbiology, Communicable Disease, Queensland Health Forensic and Scientific Services, Archerfield, Queensland, Australia..
The University of Queensland, UQ Centre for Clinical Research (UQCCR), Herston, Queensland 4029, Australia.. Pathology Queensland, Microbiology Department, Herston, Queensland, Australia..
Issue Date: 2016
Citation: The Journal of antimicrobial chemotherapy 2016; 71(11): 3090-3095
Abstract: 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). 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). 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. 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.
URI: http://docs.prosentient.com.au/prosentientjspui/handle/1/10498
DOI: 10.1093/jac/dkw291
Type: Evaluation Studies
Journal Article
Research Support, Non-U.S. Gov't
Subjects: Anti-Bacterial Agents
Australia
Genotyping Techniques
Humans
Microbial Sensitivity Tests
Neisseria gonorrhoeae
Oligonucleotide Probes
Penicillins
Porins
Predictive Value of Tests
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
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