Clinical Validation of a Real-Time Polymerase Chain Reaction Detection of Neisseria gonorrheae porA Pseudogene Versus Culture Techniques

Hjelmevoll, Stig Ove; Olsen, Merethe Elise; Sollid, Johanna U. Ericson; Haaheim, Håkon; Melby, K; Moi, Harald; Unemo, Magnus; Skogen, Vegard

doi:10.1097/olq.0b013e3181644bc9

Cited by 21 publications

(27 citation statements)

References 32 publications

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“…The low number of gonorrhea cases and the lack of large sexual networks may also suggest effective contact tracing, which is a main focus in Norway. Nevertheless, a previous study [25] and recent surveillance reports (unpublished data) have suggested that the culturebased diagnostics results in false-negative tests in Norway. This is especially true for rectal samples in MSM who practice exclusively receptive anal sex.…”

Section: Discussionmentioning

confidence: 99%

Phenotypic and genotypic properties of Neisseria gonorrhoeae isolates in Norway in 2009: antimicrobial resistance warrants an immediate change in national management guidelines

Hjelmevoll

Golparian

Dedi

et al. 2011

Eur J Clin Microbiol Infect Dis

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Despite rapidly diminishing treatment options for Neisseria gonorrhoeae and high levels of ciprofloxacin resistance worldwide, Norwegian guidelines still recommend ciprofloxacin as empirical treatment for gonorrhea. The present study aimed to characterize phenotypical and genotypical properties of N. gonorrhoeae isolates in Norway in 2009. All viable N. gonorrhoeae isolates (n = 114) from six university hospitals in Norway (2009) were collected, representing 42% of all notified gonorrhea cases. Epidemiological data were collected from the Norwegian Surveillance System for Communicable Diseases and linked to phenotypical and genotypical characteristics for each N. gonorrhoeae isolate. Resistance levels to the antimicrobials examined were: ciprofloxacin 78%, azithromycin 11%, cefixime 3.5%, ceftriaxone 1.8%, and spectinomycin 0%. The minimum inhibitory concentrations of gentamicin varied from 1.5 to 8 mg/L. Forty-one (36%) of the isolates were β-lactamase-producing, 17 displayed penA mosaic alleles, and 72 different N. gonorrhoeae multiantigen sequence types (ST; 37 novel) were identified. The most common ST was ST1407 (n = 11), containing penA mosaic allele. Four of these isolates displayed intermediate susceptibility/resistance to cefixime. The N. gonorrhoeae strains circulating in Norway were highly diverse. The level of ciprofloxacin resistance was high and the Norwegian management guidelines should promptly exclude ciprofloxacin as an empirical treatment option for gonorrhea.

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Section: Discussionmentioning

confidence: 99%

Phenotypic and genotypic properties of Neisseria gonorrhoeae isolates in Norway in 2009: antimicrobial resistance warrants an immediate change in national management guidelines

Hjelmevoll

Golparian

Dedi

et al. 2011

Eur J Clin Microbiol Infect Dis

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“…For elucidation of the spectinomycin resistance determinant, the full-length 16S rRNA gene (1,545 bp) was sequenced (51), which surprisingly revealed a wild-type gene. Consequently, the full-length rpsE gene (519 bp) was PCR amplified and sequenced with the primers 5S-F (5=-TGGCAAAACATGA AATTGAAG-3=) and 5S-R (5=-GCCATGGTTAACTCCCAAAA-3=), which were designed based on the genome sequence of the spectinomycin-susceptible gonococcal reference strain FA1090 (GenBank accession no.…”

Section: Figmentioning

confidence: 99%

Neisseria gonorrhoeae Strain with High-Level Resistance to Spectinomycin Due to a Novel Resistance Mechanism (Mutated Ribosomal Protein S5) Verified in Norway

Unemo

Golparian

Skogen

et al. 2013

Antimicrob Agents Chemother

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Gonorrhea may become untreatable, and new treatment options are essential. Verified resistance to spectinomycin is exceedingly rare. However, we describe a high-level spectinomycin-resistant (MIC, >1,024 g/ml) Neisseria gonorrhoeae strain from Norway with a novel resistance mechanism. The resistance determinant was a deletion of codon 27 (valine) and a K28E alteration in the ribosomal protein 5S. The traditional spectinomycin resistance gene (16S rRNA) was wild type. Despite this exceedingly rare finding, spectinomycin available for treatment of ceftriaxone-resistant urogenital gonorrhea would be very valuable.

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“…This is Ͻ100%, as is expected given the lower sensitivity of culture compared to that of NAAT, which is demonstrated by our direct culture positivity rate of 82% and in previous studies (4,(18)(19)(20)(21)(22)(23). The lower success of deferred cultures compared to that of direct cultures can be explained by the decreased viability of N. gonorrhoeae over time.…”

Section: Discussionmentioning

confidence: 44%

Successful Combination of Nucleic Acid Amplification Test Diagnostics and Targeted Deferred Neisseria gonorrhoeae Culture

et al. 2015

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g Nucleic acid amplification tests (NAATs) are recommended for the diagnosis of N. gonorrhoeae infections because of their superior sensitivity. Increasing NAAT use causes a decline in crucial antimicrobial resistance (AMR) surveillance data, which rely on culture. We analyzed the suitability of the ESwab system for NAAT diagnostics and deferred targeted N. gonorrhoeae culture to allow selective and efficient culture based on NAAT results. We included patients visiting the STI Clinic Amsterdam, The Netherlands, in 2013. Patient characteristics and urogenital and rectal samples for direct N. gonorrhoeae culture, standard NAAT, and ESwab were collected. Standard NAAT and NAAT on ESwab samples were performed using the Aptima Combo 2 assay for N. gonorrhoeae and C. trachomatis. Two deferred N. gonorrhoeae cultures were performed on NAAT-positive ESwab samples after storage at 4°C for 1 to 3 days. We included 2,452 samples from 1,893 patients. In the standard NAAT, 107 samples were N. gonorrhoeae positive and 284 were C. trachomatis positive. The sensitivities of NAAT on ESwab samples were 83% (95% confidence interval [CI], 75 to 90%) and 87% (95% CI, 82 to 90%), respectively. ESwab samples were available for 98 of the gonorrhea-positive samples. Of these, 82% were positive in direct culture and 69% and 56% were positive in the 1st and 2nd deferred cultures, respectively (median storage times, 27 and 48 h, respectively). Deferred culture was more often successful in urogenital samples or when the patient had symptoms at the sampling site. Deferred N. gonorrhoeae culture of stored ESwab samples is feasible and enables AMR surveillance. To limit the loss in NAAT sensitivity, we recommend obtaining separate samples for NAAT and deferred culture.A ntibiotic treatment for gonorrhea has existed for around 80 years, and Neisseria gonorrhoeae has developed mechanisms of antimicrobial resistance (AMR) ever since. Resistance to the last-resort empirical monotherapy, extended-spectrum cephalosporins, has now emerged, and this has been recognized as a major public health problem (1). In response, the World Health Organization recommended actions to extend the existing surveillance and, where lacking, develop new AMR surveillance worldwide (2, 3).In recent years, nucleic acid amplification tests (NAATs) have rapidly replaced the use of culture as a diagnostic test for gonorrhea (4). While the use of highly sensitive NAATs results in the detection of more infections, it also results in fewer cultures (5). This compromises AMR surveillance, as molecular methods to detect AMR in NAAT samples are still suboptimal, and culture remains essential (6, 7).Ideally, an NAAT to diagnose gonorrhea would be combined with targeted deferred culture of positive samples for AMR surveillance. Using a single sample for NAAT and deferred culture requires special conditions for the collection medium, i.e., maintained diagnostic NAAT sensitivity and N. gonorrhoeae survival until the NAAT result is available. The ESwab system (Copan Italia, Brescia, Ita...

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Clinical Validation of a Real-Time Polymerase Chain Reaction Detection of Neisseria gonorrheae porA Pseudogene Versus Culture Techniques

Abstract: The present porA pseudogene real-time PCR comprises a valuable supplement to the traditional culture techniques for diagnosis of N. gonorrheae, especially for samples from extragenital sites such as pharynx and rectum.

Cited by 21 publications

References 32 publications

Phenotypic and genotypic properties of Neisseria gonorrhoeae isolates in Norway in 2009: antimicrobial resistance warrants an immediate change in national management guidelines

Phenotypic and genotypic properties of Neisseria gonorrhoeae isolates in Norway in 2009: antimicrobial resistance warrants an immediate change in national management guidelines

Neisseria gonorrhoeae Strain with High-Level Resistance to Spectinomycin Due to a Novel Resistance Mechanism (Mutated Ribosomal Protein S5) Verified in Norway

Successful Combination of Nucleic Acid Amplification Test Diagnostics and Targeted Deferred Neisseria gonorrhoeae Culture

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