The influence of target population on nonculture-based detection of markers of Neisseria gonorrhoeae antimicrobial resistance

Sexual Health ◽  
2012 ◽  
Vol 9 (5) ◽  
pp. 422 ◽  
Author(s):  
Namraj Goire ◽  
Kevin Freeman ◽  
Stephen B. Lambert ◽  
Graeme R. Nimmo ◽  
Athena E. Limnios ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Direct Detection ◽  
Treatment Options ◽  
Target Population ◽  
Penicillin Binding Protein ◽  
Highly Sensitive ◽  
Genetic Mechanisms ◽  
Resistant Strains ◽  
Pcr Assays

Background With treatment options for gonorrhoea (Neisseria gonorrhoeae) diminishing, strengthening antimicrobial resistance (AMR) surveillance is paramount. Methods: In this study, we investigated polymerase chain reaction (PCR) based methods, in parallel with N. gonorrhoeae multi-antigen sequence typing (NG-MAST), for direct detection of four N. gonorrhoeae chromosomal mechanisms associated with emerging resistance to extended spectrum cephalosporins using noncultured samples: an adenine deletion in the mtrR promoter, a mosaic penicillin-binding protein (PBP) 2, an A501V PBP2 mutation, and alterations at positions 120 and 121 of the porB protein. The PCR assays were validated using a panel of characterised N. gonorrhoeae isolates (n = 107) and commensal Neisseria (n = 100) species. These PCR assays with NG-MAST were then applied to noncultured clinical specimens from distinct populations in Australia with differing levels of N. gonorrhoeae AMR: the Northern Territory (NT), where resistance has a low population prevalence, and Queensland (Qld), with higher AMR prevalence. Results: The real-time PCR assays proved highly sensitive and specific. When applied to the noncultured samples, only 1 out of 50 (2%) samples from NT harboured a resistant mechanism, whereas the Qld samples (n = 129) collected over different periods showed progressive acquisition of resistant mechanisms, and these were associated with specific NG-MAST types, including Type 225. Conclusions: The results suggest that our PCR-based methods could be used to rapidly pinpoint incursion of resistant strains into previously unaffected populations. Likewise, our results show that for molecular AMR surveillance, the population being investigated is as important as the genetic mechanisms being targeted.

scholarly journals Genomic analysis and antimicrobial resistance of Neisseria gonorrhoeae isolates from Vietnam in 2011 and 2015–16

2020 ◽  
Vol 75 (6) ◽  
pp. 1432-1438 ◽  
Author(s):  
Pham Thi Lan ◽  
Daniel Golparian ◽  
Johan Ringlander ◽  
Le Van Hung ◽  
Nguyen Van Thuong ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Reproductive Health ◽  
Neisseria Gonorrhoeae ◽  
Molecular Epidemiology ◽  
Genomic Analysis ◽  
Illumina Miseq ◽  
Phylogenomic Analysis ◽  
Asian Countries ◽  
Resistant Strains ◽  
A Minor

Abstract Objectives Antimicrobial resistance (AMR) in Neisseria gonorrhoeae, compromising gonorrhoea treatment, is a threat to reproductive health globally. South-East and East Asia have been major sources of emergence and subsequent international spread of AMR gonococcal strains during recent decades. We investigated gonococcal isolates from 2011 and 2015–16 in Vietnam using AMR testing, WGS and detection of AMR determinants. Methods Two hundred and twenty-nine gonococcal isolates cultured in 2015–16 (n = 121) and 2011 (n = 108) in Vietnam were examined. AMR testing was performed using Etest and WGS with Illumina MiSeq. Results Resistance among the 2015–16 isolates was as follows: ciprofloxacin, 100%; tetracycline, 79%; benzylpenicillin, 50%; cefixime, 15%; ceftriaxone, 1%; spectinomycin, 0%; and 5% were non-WT to azithromycin. Eighteen (15%) isolates were MDR. The MIC range for gentamicin was 2–8 mg/L. Among the 2015–16 isolates, 27% (n = 33) contained a mosaic penA allele, while no isolates had a mosaic penA allele in 2011. Phylogenomic analysis revealed introduction after 2011 of two mosaic penA-containing clones (penA-10.001 and penA-34.001), which were related to cefixime-resistant strains spreading in Japan and Europe, and a minor clade (eight isolates) relatively similar to the XDR strain WHO Q. Conclusions From 2011 to 2015–16, resistance in gonococci from Vietnam increased to all currently and previously used antimicrobials except ceftriaxone, spectinomycin and tetracycline. Two mosaic penA-containing clones were introduced after 2011, explaining the increased cefixime resistance. Significantly increased AMR surveillance, antimicrobial stewardship and use of WGS for molecular epidemiology and AMR prediction for gonococcal isolates in Vietnam and other Asian countries are crucial.

scholarly journals Antimicrobial Resistance and Molecular Typing of Neisseria gonorrhoeae Isolates in Kyoto and Osaka, Japan, 2010 to 2012: Intensified Surveillance after Identification of the First Strain (H041) with High-Level Ceftriaxone Resistance

2013 ◽  
Vol 57 (11) ◽  
pp. 5225-5232 ◽  
Author(s):  
Ken Shimuta ◽  
Magnus Unemo ◽  
Shu-ichi Nakayama ◽  
Tomoko Morita-Ishihara ◽  
Misato Dorin ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Molecular Typing ◽  
Multilocus Sequence Typing ◽  
Binding Protein ◽  
Sporadic Case ◽  
Penicillin G ◽  
Penicillin Binding Protein ◽  
Content Type ◽  
High Level

ABSTRACTIn 2009, the first high-level ceftriaxone-resistantNeisseria gonorrhoeaestrain (H041) was isolated in Kyoto, Japan. The present study describes an intensified surveillance (antimicrobial resistance and molecular typing) ofNeisseria gonorrhoeaeisolates in Kyoto and its neighboring prefecture Osaka, Japan, in 2010 to 2012, which was initiated after the identification of H041. From April 2010 to March 2012, 193N. gonorrhoeaeisolates were collected and the MICs (μg/ml) to six antimicrobials, including ceftriaxone, were determined. All isolates showed susceptibility to ceftriaxone and cefixime (MIC values, <0.5 μg/ml), and spectinomycin. The rates of resistance (intermediate susceptibility) to azithromycin, penicillin G, and ciprofloxacin were 3.6% (19.7%), 24.4% (71.0%), and 78.2% (0.5%), respectively. Multilocus sequence typing (MLST) showed that 40.9%, 19.2%, and 17.1% of isolates belonged to ST1901, ST7359, and ST7363, respectively. Furthermore,N. gonorrhoeaemultiantigen sequence typing (NG-MAST) revealed that 12 (63%) of the 19 isolates with decreased susceptibility to ceftriaxone (MIC > 0.064 μg/ml) were of ST1407. NG-MAST ST1407 was also the most prevalent ST (16.1%; 31 of 193 isolates). In those NG-MAST ST1407 strains, several mosaic typepenAalleles were found, including SF-A type (penicillin binding protein 2 allele XXXIV) and its derivatives. These were confirmed using transformation of thepenAmosaic alleles as critical determinants for enhanced cefixime and ceftriaxone MICs. The intensified surveillance in Kyoto and Osaka, Japan, did not identify any dissemination of the high-level ceftriaxone-resistantN. gonorrhoeaestrain H041, suggesting that H041 might have caused only a sporadic case and has not spread further.

scholarly journals Antimicrobial Resistance of Neisseria gonorrhoeae and Emerging Ciprofloxacin Resistance in The Netherlands, 1991 to 1998

2000 ◽  
Vol 44 (11) ◽  
pp. 3184-3185 ◽  
Author(s):  
Albert J. de Neeling ◽  
Marga van Santen-Verheuvel ◽  
Joke Spaargaren ◽  
Rob J. L. Willems
Keyword(s):  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
The Netherlands ◽  
Neisseria Gonorrhoeae ◽  
Far East ◽  
The Far East ◽  
Resistant Strains ◽  
Ciprofloxacin Resistance ◽  
High Level

ABSTRACT Surveillance of antibiotic resistance in Neisseria gonorrhoeae showed a decrease in the percentage of β-lactamase-producing isolates but an increase in intermediately penicillin-resistant strains and strains resistant to a high level of tetracycline. MICs for the ciprofloxacin-resistant isolates that emerged increased, and these isolates had mutations in gyrAand parC similar to those observed in the Far East.

scholarly journals A Novel Mechanism of High-Level, Broad-Spectrum Antibiotic Resistance Caused by a Single Base Pair Change in Neisseria gonorrhoeae

mBio ◽  
2011 ◽  
Vol 2 (5) ◽  
Author(s):  
Elizabeth A. Ohneck ◽  
Yaramah M. Zalucki ◽  
Paul J. T. Johnson ◽  
Vijaya Dhulipala ◽  
Daniel Golparian ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Base Pair ◽  
Health Concern ◽  
Content Type ◽  
Single Base ◽  
Single Base Pair ◽  
Resistant Strains ◽  
High Level

ABSTRACTThe MtrC-MtrD-MtrE multidrug efflux pump ofNeisseria gonorrhoeaeconfers resistance to a diverse array of antimicrobial agents by transporting these toxic compounds out of the gonococcus. Frequently in gonococcal strains, the expression of themtrCDEoperon is differentially regulated by both a repressor, MtrR, and an activator, MtrA. ThemtrRgene lies 250 bp upstream of and is transcribed divergently from themtrCDEoperon. Previous research has shown that mutations in themtrRcoding region and in themtrR-mtrCDEintergenic region increase levels of gonococcal antibiotic resistance andin vivofitness. Recently, a C-to-T transition mutation 120 bp upstream of themtrCstart codon, termedmtr120, was identified in strain MS11 and shown to be sufficient to confer high levels of antimicrobial resistance when introduced into strain FA19. Here we report that this mutation results in a consensus −10 element and that its presence generates a novel promoter formtrCDEtranscription. This newly generated promoter was found to be stronger than the wild-type promoter and does not appear to be subject to MtrR repression or MtrA activation. Although rare, themtr120mutation was identified in an additional clinical isolate during sequence analysis of antibiotic-resistant strains cultured from patients with gonococcal infections. We propose thatcis-acting mutations can develop in gonococci that significantly alter the regulation of themtrCDEoperon and result in increased resistance to antimicrobials.IMPORTANCEGonorrhea is the second most prevalent sexually transmitted bacterial infection and a worldwide public health concern. As there is currently no vaccine againstNeisseria gonorrhoeae, appropriate diagnostics and subsequent antibiotic therapy remain the primary means of infection control. However, the effectiveness of antibiotic treatment is constantly challenged by the emergence of resistant strains, mandating a thorough understanding of resistance mechanisms to aid in the development of new antimicrobial therapies and genetic methods for antimicrobial resistance testing. This study was undertaken to characterize a novel mechanism of antibiotic resistance regulation inN. gonorrhoeae. Here we show that a single base pair mutation generates a second, stronger promoter formtrCDEtranscription that acts independently of the known efflux system regulators and results in high-level antimicrobial resistance.

A multiplex molecular assay for detection of six penA codons to predict decreased susceptibility to cephalosporins in Neisseria gonorrhoeae

2022 ◽  
Author(s):  
Yamei Li ◽  
Lulu Zhang ◽  
Leshan Xiu ◽  
Di Wang ◽  
Yaling Zeng ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
High Throughput ◽  
High Resolution Melting ◽  
Clinical Samples ◽  
Nucleotide Polymorphisms ◽  
Rapid Testing ◽  
Single Nucleotide ◽  
Specificity And Sensitivity ◽  
Resistant Strains

The emerging cephalosporin-resistant Neisseria gonorrhoeae poses an urgent threat to the continued efficacy of the last-line monotherapy for gonorrhea. Consequently, high-throughput, accurate, and reasonable molecular assays are urgently needed for strengthening antimicrobial-resistance surveillance in N. gonorrhoeae . In this study, we designed a high-throughput multiplex method that incorporates high-resolution melting technology and is based on a 6-codon assay (among the most parsimonious assays) developed following comprehensive and systematic reviews. The results showed that our method can precisely distinguish specific single-nucleotide polymorphisms in resistance-associated genes with a specificity and sensitivity of 100% and a detection limit as low as 10 copies per reaction. This method can be directly applied to clinical samples without cumbersome culture and successfully predicted all cephalosporin-resistant isolates (sensitivity: 100%). The method presented here represents a technique for rapid testing of antimicrobial resistance and will serve as a valuable tool for tailor-made antimicrobial therapy and for monitoring the transmission of cephalosporin-resistant strains.

scholarly journals Effect of Variants of Penicillin-Binding Protein 2 on Cephalosporin and Carbapenem Susceptibilities in Neisseria gonorrhoeae

2015 ◽  
Vol 59 (8) ◽  
pp. 5003-5006 ◽  
Author(s):  
Amrita Bharat ◽  
Walter Demczuk ◽  
Irene Martin ◽  
Michael R. Mulvey
Keyword(s):  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Binding Protein ◽  
Penicillin Binding Protein ◽  
Wild Type ◽  
Molecular Surveillance ◽  
Content Type ◽  
Extended Spectrum ◽  
The Relationship

ABSTRACTTo characterize the relationship between penicillin-binding protein 2 (PBP2/penA) and susceptibility to extended-spectrum cephalosporins (ESCs) and carbapenem antibiotics, we compared 17 PBP2 variants inNeisseria gonorrhoeae. Nonmosaic and mosaic variants of PBP2 caused decreased susceptibility to ESCs and, to a lesser extent, to carbapenems. An A501P substitution in mosaic XXXIV_A501P conferred decreased susceptibility to ESCs but restored carbapenem susceptibility to wild-type levels. These results could aid the molecular surveillance of antimicrobial resistance to these agents.

scholarly journals Multitarget PCR Assay for Direct Detection of Penicillinase-Producing Neisseria gonorrhoeae for Enhanced Surveillance of Gonococcal Antimicrobial Resistance

2015 ◽  
Vol 53 (8) ◽  
pp. 2706-2708 ◽  
Author(s):  
Cameron Buckley ◽  
Ella Trembizki ◽  
Robert W. Baird ◽  
Marcus Chen ◽  
Basil Donovan ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Direct Detection ◽  
False Negative ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Content Type ◽  
Negative Results ◽  
False Negative Results ◽  
Sequence Variations

A multitarget PCR was developed for the direct detection of penicillinase-producing Neisseria gonorrhoeae (PPNG). The assay was validated by testing 342 PPNG isolates and 415 clinical samples. The method is suitable for routine detection of PPNG strains. Its multitarget approach reduces the potential for false-negative results caused by sequence variations.

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