Ciprofloxacin-resistant Neisseria Gonorrhoeae Research Articles

Genomic analyses of ciprofloxacin-resistant Neisseria gonorrhoeae isolates recovered from the largest South American metropolitan area.

We sequenced 13Neisseria gonorrhoeae isolates exhibiting distinct susceptibility profiles and which were recovered over 12years in the metropolitan region of São Paulo, Brazil. Whole Genome Sequencing (WGS) was performed on an Illumina MiSeq™2×300bp paired-end reads. Bioinformatics analyses were carried out using CGE, PATRIC, and BLAST databases for manual curation of obtained genomes. Multilocus sequence typing (MLST) analysis identified seven STs, namely ST1580, ST1590, ST1901, ST1902, ST8161, ST9363, and ST15640. Moreover, a diversity of mutations was observed in MtrR/G45D-A39T, PIB/G120K-A121S, and PBP1/L421P. Mutations associated with sulfonamides (DHPS/R228S) and rifampicin (RNAP/H552N) were also detected, as well as tetracycline resistance determinants, namely rpsJ/V57M and tet(M). The results presented herein can contribute to the knowledge of N. gonorrhoeae strains circulating in Sao Paulo, Brazil.

1438. Dissecting the Multifaceted Nature of Antibiotic Resistance in Clinical Isolates of Neisseria gonorrhoeae by Natural Transformation

Background Neisseria gonorrhoeae (NG) causes the sexually transmitted disease gonorrhea. It has developed resistance to every antibiotic introduced for gonorrhea treatment such that NG clinical isolates with multidrug resistance (MDR) are increasingly common. We hypothesize that natural transformation could be used to transfer genetic determinants of antibiotic resistance from drug-resistant NG clinical isolates without pre-knowledge of the genetic determinants to a new background under antibiotic selection to generate isogenic transformants for further characterization.MethodsNatural transformation, PCR amplification and DNA sequencing, and antibiotic susceptibility testing were used in the studies.ResultsWe have validated the hypothesis using genomic DNA from an MDR including ciprofloxacin-resistant NG clinical isolate as a donor and a ciprofloxacin-susceptible NG isolate as a recipient under the selective pressure of ciprofloxacin. This led to a series of transformants that contain single or multiple genetic resistance determinants being generated depending on the resistance levels and transformation frequencies. Antibiotic susceptibility testing and genetic characterization of the transformants allowed us to (i) identify ciprofloxacin resistance determinants including efflux mutation mtrR-79 and target mutations GyrA S91F D95G and ParC D86N, (ii) quantify the contribution of each genetic determinant responsible for the ciprofloxacin resistance and (iii) regenerate the ciprofloxacin resistance phenotype of the donor isolate to detect multiple paths of possible resistance development for ciprofloxacin. Furthermore, we also validated the hypothesis with a novel antibiotic gepotidacin to identify pre-existing genetic determinants contributing to varying susceptibility to this antibiotic. Finally, we envision that, along with whole genome sequencing, natural transformation could be used to identify and quantify novel genetic resistance determinants to current or novel antibiotics in drug-resistant NG clinical isolates.ConclusionWe demonstrated the utility of natural transformation in dissecting the multifaceted nature of antibiotic resistance in NG clinical isolates.DisclosuresJianzhong Huang, PhD, GlaxoSmithKline (Employee, Shareholder) Karen Ingraham, MS, GlaxoSmithKline (Employee, Shareholder) Pan Chan, PhD, GlaxoSmithKline (Employee, Shareholder) Steve Rittenhouse, PhD, GlaxoSmithKline (Employee, Shareholder)

Sharp increase in ciprofloxacin resistance of Neisseria gonorrhoeae in Yaounde, Cameroon: analyses of a laboratory database period 2012-2018.

A previous study conducted in Yaoundé, Cameroon, showed a significant increase in ciprofloxacin-resistant Neisseria gonorrhoeae. Still, ciprofloxacin is the recommended first-line medication for gonorrhoea in Cameroon. We aimed to provide further evidence of increasing resistance of N. gonorrhoeae to ciprofloxacin. We conducted a retrospective study (2012–2018) and retrieved coded patient and specimen real-world data from the Centre Pasteur du Cameroun’s laboratory information system and laboratory worksheets. We included disc diffusion method results for tetracycline, azithromycin, and spectinomycin, and minimal inhibitory concentrations obtained by E-test for ciprofloxacin, ceftriaxone, and benzylpenicillin. A total of 449 N. gonorrhoeae were isolated; a higher proportion of them presented resistance to ciprofloxacin (64.4%), benzylpenicillin (80.1%), and tetracycline (58.4%). The resistance to ciprofloxacin increased significantly (p < 0.0001) from 15.0% in 2012 to 79.5% in 2018. Since 2016 a significant decrease (p = 0.002) in resistance to benzylpenicillin occurred and the resistance to tetracycline remained stable. The level of resistance to ceftriaxone (1.8%), azithromycin (2.1%), and spectinomycin (2.0%) was low. Our study confirms that ciprofloxacin is not recommended as a first-line treatment for gonorrhoea. It also illustrates the urgent need to establish a robust and sustainable gonococcal antimicrobial surveillance programme to guide national treatment guidelines in order to control gonorrhoea in Cameroon.

Can Ciprofloxacin be Used for Precision Treatment of Gonorrhea in Public STD Clinics? Assessment of Ciprofloxacin Susceptibility and an Opportunity for Point-of-Care Testing.

Background: Given the lack of new antimicrobials to treat Neisseria gonorrhoeae (NG) infections, reusing previously recommended antimicrobials has been proposed as a strategy to control the spread of multi-drug-resistant NG. We assessed ciprofloxacin susceptibility in a large sample set of NG isolates and identified correlates associated with ciprofloxacin-resistant NG infections. Methods: NG isolates collected in Baltimore, Maryland between 2014 and 2016 were evaluated by Gyrase A (gyrA) PCR and E-test for susceptibility to ciprofloxacin. Clinical characteristics and demographics were evaluated by multivariate regression analysis to identify correlates of ciprofloxacin-resistant NG infections. Results: 510 NG isolates from predominately African American (96.5%), heterosexual (85.7%), and HIV-negative (92.5%) male subjects were included in the study. The overall percentage of isolates with mutant gyrA sequences, indicative of ciprofloxacin resistance, was 32.4%, and significantly increased from 24.7% in 2014 to 45.2% in 2016 (p < 0.001). Participants older than 35 years of age were 2.35 times more likely to have a gyrA mutant NG infection than younger participants (p < 0.001). Race, sexual orientation, symptomology, or co-infection the HIV or syphilis were not associated with a particular NG gyrA genotype. Conclusions: Resistance to ciprofloxacin in Baltimore is lower than other regions and indicates that in this environment, use of ciprofloxacin may be appropriate for targeted treatment provided utilization of enhanced surveillance tools. The targeted use of ciprofloxacin may be more beneficial for individuals under 35 years of age. Point-of-care tests for NG diagnosis and susceptibility testing are urgently needed to identify individuals who can be treated with this targeted approach.

Emergence and spread of ciprofloxacin-resistant Neisseria gonorrhoeae in New South Wales, Australia: lessons from history.

Our aim was to investigate the emergence and spread of ciprofloxacin resistance in clinical Neisseria gonorrhoeae isolates in New South Wales, Australia, from the first reported case in 1991 until ciprofloxacin resistance was sustained at or above the WHO threshold for treatment change of 5% (1999), to inform future strategies for controlling gonococcal antimicrobial resistance. The index isolate and all subsequent clinical isolates of ciprofloxacin-resistant N. gonorrhoeae in New South Wales from 1991 to 1999 were genotyped using a previously described method on the Agena MassARRAY iPLEX platform. Region of acquisition data, where available, were used to determine whether cases were travel associated. In New South Wales, of the 325 ciprofloxacin-resistant N. gonorrhoeae isolates reported from 1991 to 1999, 98% (320/325) were able to be recovered and 100% (320/320) were genotyped. There were 66 different genotypes, comprising 1-99 isolates each. Notably no single clone was found to account for ciprofloxacin resistance being sustained in the population, with considerable variability in genotype prevalence observed throughout the study period. A total of 65% (209/320) of genotyped isolates had information regarding the likely place of acquisition; of these, 44% (93/209) were associated with overseas travel or sexual contact with an overseas visitor. The first ciprofloxacin-resistant N. gonorrhoeae in New South Wales was associated with travel to Thailand. Index cases of each resistant genotype were significantly more likely to have been acquired overseas (51.5%), predominantly in Asia (45%, 30/66). The continued importation of multiple genotypes, rather than the expansion of a single genotype, led to ciprofloxacin-resistant N. gonorrhoeae being established in New South Wales.

First description of a cefixime- and ciprofloxacin-resistant Neisseria gonorrhoeae isolate with mutations in key antimicrobial susceptibility-determining genes from the country of Georgia.

Antimicrobial resistance in Neisseria gonorrhoeae is a global health problem. Enhanced international collaborative surveillance and disease control are needed to reduce the global burden of this important pathogen. Currently the antimicrobial resistance properties and molecular mechanisms of multidrug-resistant N. gonorrhoeae in the Republic of Georgia represent a significant knowledge gap. Here we report the isolation of a strain of N. gonorrhoeae exhibiting resistance to cefixime and ciprofloxacin with reduced susceptibility to penicillin and tetracycline from a patient being treated at a Georgian medical centre. Notably, this isolate was found to contain a mosaic penA allele and to harbour mutations in genes conferring susceptibility to the β-lactam, cephalosporin, fluoroquinolone, macrolide and penicillin classes of antibiotic. To our knowledge, this is the first report to describe the key mutations conferring the antimicrobial resistance properties of an isolate of N. gonorrhoeae from Georgia.

P1-S1.10 Microbiological surveillance for sexually transmitted infections in Windhoek and Oshakati, Namibia (2007)

BackgroundA microbiological survey was undertaken to enable review of Namibia's syndromic management-based STI treatment guidelines (1999).MethodsThis Government-approved survey took place at Katutura Health Centre (Windhoek) and Oshakati Intermediate Hospital over...

Escalation in the relative prevalence of ciprofloxacin-resistant gonorrhoea among men with urethral discharge in two South African cities: association with HIV seropositivity

Objectives:The objectives of this study were to assess the prevalence of ciprofloxacin-resistant gonorrhoea in two South African cities and to investigate the association between the isolation of ciprofloxacin-resistant Neisseria gonorrhoeae...

Mutations in gyrA and parC in ciprofloxacin-resistant Neisseria gonorrhoeae in Kuala Lumpur, Malaysia

We read with interest the recent report on quinolone-resistant Neisseria gonorrhoeae in Shanghai.1 Ciprofloxacin resistance in N. gonorrhoeae is a serious problem worldwide, especially in Southeast Asia.2 In Malaysia, resistance rates to ciprofloxacin, penicillin and tetracycline have previously been reported to be . 50%.2 At our teaching hospital in Kuala Lumpur, Malaysia, ciprofloxacin resistance rates were 41.4% in 2006... (extract)

Optimizing Treatment of Antimicrobial-resistantNeisseria gonorrhoeae

The increasing prevalence of ciprofloxacin-resistant Neisseria gonorrhoeae has required replacing inexpensive oral ciprofloxacin treatment with more expensive injectable ceftriaxone. Further, monitoring antimicrobial resistance requires culture testing, but nonculture gonorrhea tests are rapidly replacing culture. Since the strategies were similar in effectiveness (> 99%), we evaluated, from the healthcare system perspective, cost-minimizing strategies for both diagnosis (culture followed by antimicrobial susceptibility tests versus nonculture-based tests) and treatment (ciprofloxacin versus ceftriaxone) of gonorrhea in women. Our results indicate that switching from ciprofloxacin to ceftriaxone is cost-minimizing (i.e., optimal) when the prevalence of gonorrhea is > 3% and prevalence of ciprofloxacin resistance is > 5%. Similarly, culture-based testing and susceptibility surveillance are optimal when the prevalence of gonorrhea is < 13%; nonculture-based testing is optimal (cost-minimizing) when gonorrhea prevalence is > or = 13%.

Fluoroquinolone Resistance Among Neisseria gonorrhoeae Isolates in Hawaii, 1990–2000

In 1999, an increase in ciprofloxacin-resistant Neisseria gonorrhoeae isolates was identified in Hawaii, prompting initiation of investigative studies. The goal of this study was epidemiologic evaluation of this increase. The authors conducted a review of laboratory data; case-series and case-control studies based on medical record review; and a prospective case-control study based on patient interviews. A total of 10.4% (21 of 201) of gonococcal isolates from Hawaii in 2000 were ciprofloxacin-resistant compared with <1.5% per year from 1990 to 1997. From medical record review for patients diagnosed with ciprofloxacin-resistant N. gonorrhoeae infection from 1990 to 1999, 59% were Asian/Pacific Islanders and 91% were heterosexual. From review of 1998 and 1999 sexually transmitted disease (STD) clinic medical records, patients with ciprofloxacin-resistant N. gonorrhoeae were more likely to report recent foreign travel or a sex partner with recent foreign travel than patients with ciprofloxacin-susceptible N. gonorrhoeae (6 of 12 vs. 10 of 117, P <0.001), but 50% (6 of 12) acquired a ciprofloxacin-resistant strain locally from a partner with no recent travel. In 2000, 70% (7 of 10) of STD clinic patients with ciprofloxacin-resistant N. gonorrhoeae acquired their infection locally from partners with no reported recent travel. Infections with ciprofloxacin-resistant N. gonorrhoeae are increasing and evolving in Hawaii.

Molecular epidemiology of endemic ciprofloxacin-resistant Neisseria gonorrhoeae in Liverpool.

Ciprofloxacin-resistant Neisseria gonorrhoeae had been rarely detected on Merseyside and when found was associated with beta-lactamase producing strains, imported from abroad. However, in August 2000, two cases of infection with ciprofloxacin-resistant beta-lactamase-negative strains occurred in sexually unrelated patients with no history of foreign travel. Over the next 18 months a total of 120 patients presented with ciprofloxacin-resistant gonococci, from which 99 patient strains were available for study. Gonococcal DNA was subjected to molecular fingerprinting by polymerase chain reaction amplification followed by Taq1 digestion of their opa genes. Twelve differing opa-types were found, but 79 patients were infected with a single genotype, opa-type 1. The sexual histories of the majority of this group indicated acquisition in Merseyside. This endemic strain was further characterized by having the same amino acid substitutions on gyrA and parC genes. An endemic clone of ciprofloxacin-resistant N. gonorrhoeae has been established on Merseyside necessitating the introduction of ceftriaxone as first-line treatment. Despite the presence of 11 other clones in the city, opa type-1 strains have not yet been displaced, raising the possibility that this strain is endowed with added virulence/endemicity traits or that a number of source patients have not yet been found.

Investigation of an outbreak of ciprofloxacin-resistant Neisseria gonorrhoeae using a simplified opa-typing method.

Ciprofloxacin-resistant gonococci have been isolated from patients in the United Kingdom since 1993. Until recently, evidence has suggested that the majority of infections are not endemic but have been acquired abroad. In October 1999, increasing numbers of ciprofloxacin resistant isolates of the non-requiring auxotype were reported in Oldham and Rochdale (Northwest England). These and similar isolates from elsewhere in England and Wales were genetically characterized using a simplified opa-typing method (a non-radioactive PCR-RFLP method targeting the opa family of genes). Of 73 isolates studied, 24 had unique opa-types (10 from infections acquired abroad), whilst the remaining 49 were indistinguishable (none were known to be acquired abroad). This cluster included 31 isolates from Oldham and Rochdale, 16 from elsewhere in the north of England, and 2 from Southern England and South Wales with known epidemiological links to cases from Manchester and Rochdale respectively. This study illustrates the potential for spread of an antibiotic resistant clone of N. gonorrhoeae both locally and nationally and demonstrates that endemic acquisition of ciprofloxacin-resistant gonococci is now a significant problem in the United Kingdom.