Relation between Genetic Markers of Drug Resistance and Susceptibility Profile of Clinical
            <i>Neisseria gonorrhoeae</i>
            Strains

Ilina, Elena N.; Vereshchagin, V. A.; Borovskaya, Alexandra D.; Malakhova, Maja V.; Сидоренко, С. В.; Al-Khafaji, Nazar; Кубанова, А. А.; Govorun, Vadim M.

doi:10.1128/aac.01420-07

Cited by 34 publications

(33 citation statements)

References 30 publications

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“…Thus, for our study, nucleic acid extraction was performed directly from specimens submitted for Aptima testing and from suspensions of cultures prepared in saline using the QIAamp DNA minikit. These extracts were used for amplification and sequencing of the penA, por, and tbpB gene targets performed using previously described primers (7,16,17) for direct molecular characterizations. In a preliminary pilot study, mutations in the ponA gene, coding for PBP 1, pilQ coding for the pilus secretin protein, bla TEM-1 coding for ␤-lactamase, and the Mtr promoter and repressor regions in addition to the penA, por, and tbpB genes were also monitored using samples with elevated cefixime MIC values; however, a reproducible association was found between changes in penA, por, and tbpB and decreased susceptibility for the limited number of samples tested in this study (data not shown).…”

Section: Methodsmentioning

confidence: 99%

Utility of Specimens Positive for Neisseria gonorrhoeae by the Aptima Combo 2 Assay for Assessment of Strain Diversity and Antibiotic Resistance

et al. 2013

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In our jurisdiction, the Aptima Combo 2 assay (Gen-Probe, Inc.) is used to detect Neisseria gonorrhoeae from specimens collected at clinics for sexually transmitted infections (STI) and from select community patients. In addition, swabs are also collected for N. gonorrhoeae culture, susceptibility testing, and sequence typing (ST). Since only a small proportion of samples from provincial cases undergo culture, the available trends in antimicrobial susceptibility and predominant strain types may not be representative of all N. gonorrhoeae infections. Due to the limitations facing the use of N. gonorrhoeae culture to understand these trends in the general community, we performed a molecular analysis for markers of cephalosporin resistance and ST determination by using nucleic acid extracts of specimens sent for Aptima testing. Thirty-four samples submitted for both Aptima testing and N. gonorrhoeae culture from the same anatomic location (within 24 h) were included in the study. Sequence type was determined based on the sequence of the por and tbpB genes, and amino acid changes in the PBP 2 protein, encoded by the penA gene, were considered representative for the assessment of antimicrobial susceptibility. Sequence identity of 100% was observed between the sequences obtained from Aptima-analyzed samples and culture samples. Sequencing results showed an association between decreased susceptibility to extended-spectrum cephalosporins (ESC ds ), tbp allele 110, ST 1407, and amino acid changes (G545S, I312M, and V316T) in the PBP 2 protein. Our data, generated based on a few representative genes, suggest that gonococcal samples positive by Aptima testing can be used to determine single nucleotide polymorphisms associated with ESC ds and the sequence type based on molecular strain typing. Confirmation of these findings may obviate the need for gonorrhea culture in the future. N eisseria gonorrhoeae is a common sexually transmitted infection that affects mucosal surfaces and causes a spectrum of conditions, including urethritis, endocervicitis, pelvic inflammatory disease, and infertility (1). Over the years, gonococci have developed resistance to multiple classes of antibiotics, including penicillins, tetracyclines, macrolides, and quinolones (2). Of recent concern is a steady increase in the MICs to extended-spectrum cephalosporins, including cefixime and ceftriaxone, in several countries including Canada, with reported levels at 0.12 g/ml, one doubling dilution away from the Clinical and Laboratory Standards Institute (CLSI) limit of 0.25 g/ml for susceptibility (3-7; CLSI standard M100-S23, January 2013). According to a recent U.S. Centers for Disease Control and Prevention (CDC) update, untreatable gonorrhea could soon be a reality in the United States (http://www.cdc.gov/mmwr/preview /mmwrhtml/mm6131a3.htm). The agency noted that N. gonorrhoeae seems to be developing resistance to the oral antibiotic cefixime, and it no longer recommends cefixime at any dose as a firstline regimen for treatment of gonococcal infec...

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Section: Methodsmentioning

confidence: 99%

Utility of Specimens Positive for Neisseria gonorrhoeae by the Aptima Combo 2 Assay for Assessment of Strain Diversity and Antibiotic Resistance

et al. 2013

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“…N. gonorrhoeae clinical isolates collected in different regions of Russia from 2004 to 2005 were obtained from the Central Research Institute of Dermatology and Venereology. All isolates were identified as N. gonorrhoeae and tested for serotype and for susceptibility to penicillin G (PEN), tetracycline (TET), ciprofloxacin (CIP), spectinomycin, and ceftriaxone as a part of a previous investigation (11).…”

Section: Methodsmentioning

confidence: 99%

“…In Russia, strains that are resistant to penicillin, tetracycline, and fluoroquinolones are common, reaching up to 60% prevalence in some regions (14). As is generally known, most resistance mechanisms in N. gonorrhoeae are linked to mutations in genomic DNA, and a wide dissemination of such mutations has been demonstrated in subjects with gonorrhea in the Russian population (11). In fact, chromosomally mediated drug resistance expands clonally in a bacterial population.…”

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confidence: 99%

Molecular Surveillance of Clinical Neisseria gonorrhoeae Isolates in Russia

Ilina¹,

Oparina²,

Shitikov³

et al. 2010

J Clin Microbiol

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The choice of adequate methods for epidemiological purposes remains a challenging problem in Neisseria gonorrhoeae molecular monitoring. In this study, the collection of geographically unrelated gonococci (n ‫؍‬ 103) isolated in Russian clinics was comparably tested by (i) a traditional serotyping scheme, (ii) por typing, (iii) Neisseria gonorrhoeae multiantigen sequence typing (NG-MAST), and (iv) multilocus sequence typing (MLST). It is shown that, according to sequencing data, a third of the strains carried new porB1 alleles, as well as tbpB ones, and more than half of the samples had new sequence types (STs) as determined by NG-MAST or MLST. The discriminatory power for each typing method was calculated by using the Hunter-Gaston discriminatory index, D. Commonly, modern nucleic acid-based typing methods (por typing, NG-MAST, and MLST) appeared to be more efficient than the classical serotyping scheme. While the traditional serotyping gave a D value of 0.82, the por typing, NG-MAST, and MLST approaches yielded D values of 0.97, 0.98, and 0.91, respectively. Each typing technique revealed the distribution of gonococci slightly correlated with their geographical sources. However, only the MLST method STs were highly associated with certain phenotypes. Although ST1594, ST1892, and ST6720 were typical for susceptible gonococci, ST1901 and ST6716 were undoubtedly associated with a multidrug-resistant phenotype. We conclude that every tested nucleic acid-based typing method is suitable for N. gonorrhoeae molecular surveillance. However, the MLST method seems to serve large-scale epidemiological purposes, whereas the NG-MAST and por typing approaches are more appropriate for the investigation of local outbreaks.

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“…Unfortunately, no commercially available gonococcal NAATs detect any AMR determinants. However, in-house molecular assays exist for detection of one or more genetic AMR resistance determinants involved in plasmid-mediated penicillin resistance (29)(30)(31), chromosomally mediated penicillin resistance (26,(32)(33)(34)(35), plasmid-and chromosomally mediated tetracycline resistance (36), resistance to macrolides (32, 37-42), fluoroquinolone resistance (43-49), ESC resistance (50)(51)(52)(53), and multidrug resistance (54)(55)(56). Unfortunately, for most AMR determinants, the sensitivity and specificity of these molecular AMR assays for determination of AMR are often low.…”

Section: Detection Of Antimicrobial Resistance In N Gonorrhoeaementioning

confidence: 99%

Antimicrobial Resistance in Neisseria gonorrhoeae in the 21st Century: Past, Evolution, and Future

2014

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SUMMARY Neisseria gonorrhoeae is evolving into a superbug with resistance to previously and currently recommended antimicrobials for treatment of gonorrhea, which is a major public health concern globally. Given the global nature of gonorrhea, the high rate of usage of antimicrobials, suboptimal control and monitoring of antimicrobial resistance (AMR) and treatment failures, slow update of treatment guidelines in most geographical settings, and the extraordinary capacity of the gonococci to develop and retain AMR, it is likely that the global problem of gonococcal AMR will worsen in the foreseeable future and that the severe complications of gonorrhea will emerge as a silent epidemic. By understanding the evolution, emergence, and spread of AMR in N. gonorrhoeae , including its molecular and phenotypic mechanisms, resistance to antimicrobials used clinically can be anticipated, future methods for genetic testing for AMR might permit region-specific and tailor-made antimicrobial therapy, and the design of novel antimicrobials to circumvent the resistance problems can be undertaken more rationally. This review focuses on the history and evolution of gonorrhea treatment regimens and emerging resistance to them, on genetic and phenotypic determinants of gonococcal resistance to previously and currently recommended antimicrobials, including biological costs or benefits; and on crucial actions and future advances necessary to detect and treat resistant gonococcal strains and, ultimately, retain gonorrhea as a treatable infection.

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Relation between Genetic Markers of Drug Resistance and Susceptibility Profile of Clinical Neisseria gonorrhoeae Strains

Cited by 34 publications

References 30 publications

Utility of Specimens Positive for Neisseria gonorrhoeae by the Aptima Combo 2 Assay for Assessment of Strain Diversity and Antibiotic Resistance

Utility of Specimens Positive for Neisseria gonorrhoeae by the Aptima Combo 2 Assay for Assessment of Strain Diversity and Antibiotic Resistance

Molecular Surveillance of Clinical Neisseria gonorrhoeae Isolates in Russia

Antimicrobial Resistance in Neisseria gonorrhoeae in the 21st Century: Past, Evolution, and Future

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