Typing and antimicrobial susceptibility of 134 Neisseria gonorrhoeae strains from Southern Spain.

OBJECTIVE: Last guidelines have recommended the introduction of dual antimicrobial therapy in order to avoid treatment failure. In the present report, the susceptibility to some antibiotics was evaluated, and the typing of Neisseria gonorrhoeae strains was performed. METHODS: Gonococcal isolates were tested for susceptibility according to the recommendations of both CLSI and EUCAST. A total of 134 isolates were typed by the NG-MAST technique. RESULTS: Seventy-two different N. gonorrhoeae types were found, and the most frequent types obtained were ST 1407, ST 14958, ST 7192, ST 13251 and ST 5405. If CLSI/EUCAST criteria were applied, a ST 9807 type was found nonsusceptible to ceftriaxone and cefixime (MIC 0.5 mg/L), and a ST 12800 type was found nonsusceptible only to cefixime (MIC 0.25 mg/L). When only EUCAST breakpoints were taken into account, three strains were also resistant to cefixime (MIC 0.25 mg/L) and three isolates were resistant to ceftriaxone (MIC 0.19, 0.16 and 0.25 mg/L, respectively). The majority of strains were resistant to ciprofloxacin (68.6%), and all N. gonorrhoeae strains were susceptible to spectinomycin; 9.7% of isolates were resistant to azithromycin. CONCLUSIONS: Molecular typing may be a useful tool to predict antimicrobial resistance. High rates of resistance to penicillin, tetracycline and ciprofloxacin were found in this area. It is highly recommended to carry out antimicrobial susceptibility in all gonorrhoea cases and to identify treatment failures to verify emerging resistance.

Implementation of a Rapid Genotypic Assay to Promote Targeted Ciprofloxacin Therapy of Neisseria gonorrhoeae in a Large Health System.

Multidrug-resistant Neisseria gonorrhoeae is a top threat to public health. In November 2015, UCLA Health introduced a rapid gyrase A (gyrA) genotypic assay for prediction of Neisseria gonorrhoeae susceptibility to ciprofloxacin. We found a significant reduction in ceftriaxone use with a concomitant increase in targeted therapy.

Ceftibuten resistance and treatment failure of Neisseria gonorrhoeae infection.

Neisseria gonorrhoeae infections have been empirically treated in Hong Kong with a single oral 400-mg dose of ceftibuten since 1997. Following anecdotal reports of the treatment failure of gonorrhea with oral extended-spectrum cephalosporins, the current study was undertaken to determine the antimicrobial susceptibility pattern and molecular characteristics of isolates of N. gonorrhoeae among patients with putative treatment failure in a sexually transmitted disease clinic setting. Between October 2006 and August 2007, 44 isolates of N. gonorrhoeae were studied from patients identified clinically to have treatment failure with empirical ceftibuten. The ceftibuten MICs for three strains were found to have been 8 mg/liter. These strains were determined by N. gonorrhoeae multiantigen sequence typing to belong to sequence type 835 (ST835) or the closely related ST2469. The testing of an additional eight archived ST835 strains revealed similarly elevated ceftibuten MICs. The penA gene sequences of these 11 isolates all had the mosaic pattern previously described as pattern X. Of note is that the ceftriaxone susceptibility results of these strains all fell within the susceptible range. It is concluded that ceftibuten resistance may contribute to the empirical treatment failure of gonorrhea caused by strains harboring the mosaic penA gene, which confers reduced susceptibility to oral extended-spectrum cephalosporins. Screening for such resistance in the routine clinical laboratory may be undertaken by the disk diffusion test. The continued monitoring of antimicrobial resistance and molecular characteristics of N. gonorrhoeae isolates is important to ensure that control and prevention strategies remain effective.

Molecular characterization of Neisseria gonorrhoeae identifies transmission and resistance of one ciprofloxacin-resistant strain.

A highly discriminative and objective genetic characterization of N. gonorrhoeae, which increases our knowledge of strain populations in different geographic areas, is crucial for the development of improved control measures. In the present study, conventional phenotypic characterization and genetic characterization by means of pulsed-field gel electrophoresis (PFGE), sequencing of the entire porB gene, N. gonorrhoeae multiantigen sequence typing (NG-MAST), and pyrosequencing of a quinolone resistance determining region (QRDR) of the gyrA gene of Swedish ciprofloxacin-resistant N. gonorrhoeae serovar IB-10 isolates (n=45) were performed. The genetic characterization identified one widely spread ciprofloxacin-resistant N. gonorrhoeae ST147 strain. In addition, isolates with slightly different genetic characteristics, which presumably reflect the ongoing evolution only, were also identified. All the isolates contained single nucleotide polymorphisms in QRDR of the gyrA gene that are highly correlated with ciprofloxacin resistance. Consequently, comprehensive characterization identified the first confirmed large domestic transmission, mainly among young heterosexuals, of one ciprofloxacin-resistant N. gonorrhoeae strain in Swedish society during 2002-2003. In conclusion, a precise, i.e. genetic, characterization for identification of individual strains is a very valuable support to the crucial active surveillance of the epidemiological characteristics and the antibiotic susceptibility of N. gonorrhoeae in the effective treatment of gonorrhoea.

[Increasing incidence of ciprofloxacin resistant gonorrhea in Sweden. Choose a correct antibiotic and follow up the treatment!]. / Ciprofloxacinresistent gonorré ökar kraftigt i Sverige. Välj rätt antibiotikum och följ upp behandlingen!

The incidence of gonorrhoea has increased in Sweden and is now three times higher than in the middle of the 1990's. A remarkable increase of ciprofloxacin resistant gonorrhoea has been reported in Stockholm and other parts of Sweden during 2003. Among men attending a clinic for homosexual men in Stockholm the ciprofloxacin resistant cases have increased from a low level to over 50% during the last year. Most of the homosexual men are exposed in Stockholm and one serotype is dominant. Also in the county of Gävleborg there has been an outbreak of ciprofloxacin resistant gonorrhoea among young heterosexual men and women. No resistance to cefixime, ceftriaxone and spectinomycin has been noted and these antibiotics are then a better first choice of treatment in a Swedish context.

Failure of azithromycin therapy in gonorrhea and discorrelation with laboratory test parameters.

BACKGROUND: Azithromycin is efficacious in the treatment of chlamydial genital tract infection but less so in gonorrhea. However, MICs of azithromycin for gonococci from previously reported azithromycin treatment failures were consistently below the 'susceptible' MIC level of 2 mg/L. GOAL OF THIS STUDY: To examine gonococci not eliminated with 1 g azithromycin therapy to establish treatment outcome/MIC correlates in gonorrhea. STUDY DESIGN: The MICs and phenotypes of gonococci isolated from five cases of treatment failure after 1 g azithromycin therapy were determined and compared with the MICs of a systematic sample of routine isolates. RESULTS: Azithromycin MICs of gonococci from five cases of failed 1 g azithromycin treatment were 0.125 or 0.25 mg/L, well within the current 'susceptible' MIC range. None of the isolates were of the mtr phenotype. The MIC90 of a systematic sample of 219 gonococcal isolates was 0.25 mg/L. CONCLUSION: The antibiotic MIC/treatment outcome correlates that are usually found in gonorrhea do not apply for azithromycin. Current MIC criteria do not accurately define susceptibility or resistance of gonococci to azithromycin and by themselves do not predict the likely outcome of therapy. Pharmacokinetic factors may decrease the predictive value of MIC data.

Increasing resistance of Neisseria gonorrhoeae in west and central Africa. Consequence on therapy of gonococcal infection.

BACKGROUND AND OBJECTIVES: Antimicrobial resistant strains of Neisseria gonorrhoeae have spread with remarkable rapidity in many African countries. Chromosomal resistance to penicillin, tetracycline, and thiamphenicol is frequent now, and reported prevalences of penicillinase-producing N. gonorrhoeae isolates vary between 15% and 80%. Plasmid-mediated tetracycline-resistant N. gonorrhoeae isolates have been observed in several African countries. GOALS: To characterize gonococcal isolates from three sites in West and Central Africa, to determine antimicrobial susceptibility patterns, to document the spread of plasmid-mediated resistance to penicillin and tetracycline in these three sites, and to discuss the consequences of rising antimicrobial resistance on the management of gonococcal infection in Africa. STUDY DESIGN: Over time, a total of 2,288 gonococcal isolates were obtained from Abidjan, Ivory Coast (1992-1993, n = 251), from Kigali, Rwanda (1988-1993, n = 952), and from Kinshasa, Zaire (1988-1990, n = 1,085). The isolates were characterized by auxotyping and serotyping. Plasmid-mediated resistance to penicillin and to tetracycline was determined. Antimicrobial susceptibility testing to ceftriaxone, ciprofloxacin, penicillin, spectinomycin, tetracycline, and thiamphenicol was performed with an agar dilution method. RESULTS: The prevalence of penicillinase-producing N. gonorrhoeae increased significantly over time from 44% to 57% in Kigali and remained stable at a high level in Abidjan (73%) and in Kinshasa (67%). The frequency of tetracycline-resistant N. gonorrhoeae increased significantly during the observation periods in all three sites: from 20% to 65% in Abidjan, from 0% to 64% in Kigali, and from 14% to 41% in Kinshasa. Chromosomal resistance to penicillin was common in Kigali and Kinshasa, and chromosomal resistance to tetracycline and thiamphenicol was frequent in all three sites. All gonococcal isolates were susceptible to ceftriaxone, ciprofloxacin, and spectinomycin. Prototrophic and proline requiring strains were predominant, and IA-6 was the most common serovar in the three sites. IB-specific serovars were more common among penicillinase-producing N. gonorrhoeae and IA-specific serovars were more frequent among tetracycline-resistant N. gonorrhoeae, but there was no evidence for a clonal spread of resistant strains. CONCLUSIONS: This study illustrates the high frequency of resistant gonococci in Africa and shows that tetracycline-resistant N. gonorrhoeae have become highly endemic in different geographic areas of the continent. The use of effective drugs is essential to reduce gonorrhea transmission. Surveillance of temporal changes in antimicrobial resistance in gonococcal strain populations should be part of sexually transmitted diseases control programs.

A one-year survey of Neisseria gonorrhoeae isolated from patients attending an east London genitourinary medicine clinic: antibiotic susceptibility patterns and patients' characteristics.

OBJECTIVES: To collect epidemiological data on gonococcal infection in an east London genitourinary medicine (GUM) clinic; to perform antibiotic susceptibility testing on Neisseria gonorrhoeae isolates and relate results to patient data; to assess the efficacy of current first-line antibiotic therapy for treating gonorrhoea. METHODS: Gonococcal isolates were collected from 113 patients attending the GUM clinic at Newham General Hospital over a one year period. Isolates (104) were tested for susceptibility to various antibiotics. Plasmid profiles were obtained for penicillinase producing gonococci (PPNG) and isolates exhibiting high-level tetracycline resistance (TRNG). Epidemiological information was collected from clinic attenders by routine note-taking. RESULTS: PPNG (16) accounted for 15% of isolates tested, only three being acquired outside the United Kingdom (U.K.). Plasmid typing showed three types of beta-lactamase-encoding plasmids were represented (2.9 MDa, 3.2 MDa and 4.4 MDa). Amongst the non-PPNG isolates, high-level chromosomal resistance to penicillin (CMRNG) was found in 3.5%, intermediate resistance in 57.5% and full susceptibility in 39%. One isolate showed decreased susceptibility to ciprofloxacin (MIC = 0.06 mg/l). Three PPNG isolates also possessed a 25.2 MDa plasmid and expressed high-level tetracycline resistance encoded by tetM. All isolates were susceptible to cefixime, cefotaxime, azithromycin and spectinomycin. Most gonorrhoea (90%) was seen in local residents. The male:female case ratio was 2:1 with homosexually-acquired gonorrhoea accounting for only 3.5% of the total. Most patients (96%) had acquired gonorrhoea in the U.K.. A past history of gonorrhoea was more frequent in male patients. Concurrent chlamydial infection was seen in 31% females and 16% males. CONCLUSIONS: The high PPNG rate supports a previous decision to change first-line therapy from amoxycillin with probenecid to ciprofloxacin. There was no evidence of clinical treatment failure with ciprofloxacin. Cefixime, cefotaxime, azithromycin and spectinomycin all appear to be suitable alternative therapies. Acquisition of gonorrhoea abroad was associated with isolates exhibiting penicillin resistance but such isolates were also obtained from patients infected locally and without a history of foreign travel.

Antimicrobial susceptibility testing of Neisseria gonorrhoeae and implications for epidemiology and therapy.

Antimicrobial susceptibility testing (AST) of Neisseria gonorrhoeae has been under development since the early days of antimicrobial agents. However, it is rarely applied to clinical isolates today. The history of the various in vitro tests to determine the susceptibility of N. gonorrhoeae to antibiotics is rich with evidence that these results predict response to therapy for almost all agents tested. Further, AST is a useful and important aspect of strain characterization and disease epidemiology in conjunction with the more specific but laborious techniques of auxotyping, serotyping, and plasmid analysis. Current technology has overcome many of the objections to AST for N. gonorrhoeae with standardization of test media and the development of an accurate disk diffusion AST method that is suited to most clinical laboratories regardless of volume or level of technical expertise. Ironically, the very low level of resistance to the current primary treatment strategy in the United States, ceftriaxone or another potent cephalosporin, makes the use of AST somewhat superfluous.

Detection and differentiation of iron-responsive avirulent mutants on Congo red agar.

Agar medium containing Congo red dye differentiates virulent and avirulent colonies of Shigella, Vibrio cholerae, Escherichia coli, and Neisseria meningitidis. Like virulent plague bacilli, wild-type cells of these species absorb the dye and produce red colonies. Mutants or colonial variants have been isolated that fail to absorb the dye and produce colorless colonies. These mutants exhibit reduced virulence in the chicken embryo model, but their virulence is enhanced by supplementation with iron. Of those species tested, only Neisseria gonorrhoeae isolates failed to grow in the presence of this dye. Inhibition of growth by Congo red may thus provide a simple means for differentiating gonococci from other Neisseria.

Rapid method for auxotyping multiple strains of Neisseria gonorrhoeae.

A rapid method for auxotyping strains was developed that uses microtiter plates. This miniplate technique enables rapid identification of major auxotypes present in clinical strains. Additional growth requirements can be identified by adding individual amino acid supplements to complete gonococcal genetic medium. Analysis of 8 clones from 40 patients revealed that 10 had more than 1 auxotype. Deoxyribonucleic acid-mediated transformation can be used to establish whether the strains with apparently more than one auxotype are defective in the same locus in each of the involved biosynthetic pathways. Selection of more than one clone is required in precise epidemiological studies.