Prediction of ceftriaxone MIC in<i>Neisseria gonorrhoeae using</i>DNA microarray technology and regression analysis

Shaskolskiy, Boris; Kandinov, Ilya; Kravtsov, Dmitry; Filippova, M. A.; Chestkov, A. V.; Solomka, V S; Кубанов, А. А.; Deryabin, D. G.; Dementieva, Ekaterina; Gryadunov, Dmitry

doi:10.1093/jac/dkab308

Cited by 10 publications

(9 citation statements)

References 35 publications

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“…To date, a few isolates with chromosomal resistance to cephalosporins have been described. The typical ceftriaxone MICs for such isolates are 0.5–1.0 mg/L ( Młynarczyk-Bonikowska et al, 2020 ; Shaskolskiy et al, 2021 ). However, the emergence of ESBLs that can hydrolyze cephalosporins may lead to a several fold increase in the ceftriaxone MIC.…”

Section: Discussionmentioning

confidence: 99%

In vitro Susceptibility to β-Lactam Antibiotics and Viability of Neisseria gonorrhoeae Strains Producing Plasmid-Mediated Broad- and Extended-Spectrum β-Lactamases

et al. 2022

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Neisseria gonorrhoeae plasmids can mediate high-level antimicrobial resistance. The emergence of clinical isolates producing plasmid β-lactamases that can hydrolyze cephalosporins, the mainstay treatment for gonorrhea, may be a serious threat. In this work, N. gonorrhoeae strains producing plasmid-mediated broad- and extended-spectrum β-lactamases (ESBLs) were obtained in vitro, and their viability and β-lactam antibiotic susceptibility were studied. Artificial pblaTEM-1 and pblaTEM-20 plasmids were constructed by site-directed mutagenesis from a pblaTEM-135 plasmid isolated from a clinical isolate. Minimum inhibitory concentration (MIC) values for a series of β-lactam antibiotics, including benzylpenicillin, ampicillin, cefuroxime, ceftriaxone, cefixime, cefotaxime, cefepime, meropenem, imipenem, and doripenem, were determined. The N. gonorrhoeae strain carrying the pblaTEM-20 plasmid exhibited a high level of resistance to penicillins and second–fourth-generation cephalosporins (MIC ≥2 mg/L) but not to carbapenems (MIC ≤0.008 mg/L). However, this strain stopped growing after 6 h of culture. The reduction in viability was not associated with loss of the plasmid but can be explained by the presence of the plasmid itself, which requires additional reproduction costs, and to the expression of ESBLs, which can affect the structure of the peptidoglycan layer in the cell membrane. Cell growth was mathematically modeled using the generalized Verhulst equation, and the reduced viability of the plasmid-carrying strains compared to the non-plasmid-carrying strains was confirmed. The cell death kinetics of N. gonorrhoeae strains without the pblaTEM-20 plasmid in the presence of ceftriaxone can be described by a modified Chick–Watson law. The corresponding kinetics of the N. gonorrhoeae strain carrying the pblaTEM-20 plasmid reflected several processes: the hydrolysis of ceftriaxone by the TEM-20 β-lactamase and the growth and gradual death of cells. The demonstrated reduction in the viability of N. gonorrhoeae strains carrying the pblaTEM-20 plasmid probably explains the absence of clinical isolates of ESBL-producing N. gonorrhoeae.

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Section: Discussionmentioning

confidence: 99%

In vitro Susceptibility to β-Lactam Antibiotics and Viability of Neisseria gonorrhoeae Strains Producing Plasmid-Mediated Broad- and Extended-Spectrum β-Lactamases

et al. 2022

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“…Notably, no cases of ineffective treatment of gonococcal infection with ceftriaxone have been recorded in Russia to date [13,24,25]. However, the identification of clinical isolates with mutations that reduce susceptibility to ceftriaxone emphasizes the need for continuous monitoring of N. gonorrhoeae antimicrobial susceptibility, particularly monitoring using the developed hydrogel droplet microarrays.…”

Section: Discussionmentioning

confidence: 99%

“…Targeted gene sequencing based on the Ion AmpliSeq™ platform (Thermo Fischer Scientific, Waltham, MA, USA) or AmpliSeq for Illumina products can be used for accurate genotyping of antimicrobial resistance determinants. An alternative technology that has successfully been proven useful for simultaneous identification of multiple specific genomic targets is matrix oligonucleotide microarray analysis, which is currently widely used [11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

“…Previously, we developed a microarray to identify 12 causative agents of human reproductive tract infections, including N. gonorrhoeae, with simultaneous detection of numerous genetic drug resistance determinants [23]. We also developed a microarray to detect numerous mutations leading to the resistance of N. gonorrhoeae to ceftriaxone, a drug of choice for the treatment of gonorrhea [13].…”

Section: Introductionmentioning

confidence: 99%

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Hydrogel Droplet Microarray for Genotyping Antimicrobial Resistance Determinants in Neisseria gonorrhoeae Isolates

et al. 2021

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A multiplex assay based on a low-density hydrogel microarray was developed to identify genomic substitutions in N. gonorrhoeae that determine resistance to the currently recommended treatment agents ceftriaxone and azithromycin and the previously used drugs penicillin, tetracycline, and ciprofloxacin. The microarray identifies 74 drug resistance determinants in the N. gonorrhoeae penA, ponA, porB, gyrA, parC, rpsJ, mtrR, blaTEM, tetM, and 23S rRNA genes. The hydrogel elements were formed by automated dispensing of nanoliter-volume droplets followed by UV-induced copolymerization of NH2-containing oligonucleotides with gel-forming monomers. Polybutylene terephthalate plates without special modifications were used as microarray substrates. Sequences and concentrations of immobilized oligonucleotides, gel composition, and hybridization conditions were carefully selected, and the median discrimination ratio ranged from 2.8 to 29.4, allowing unambiguous identification of single-nucleotide substitutions. The mutation identification results in a control sample of 180 N. gonorrhoeae isolates were completely consistent with the Sanger sequencing results. In total, 648 clinical N. gonorrhoeae isolates obtained in Russia during the last 5 years were analyzed and genotyped using these microarrays. The results allowed us to draw conclusions about the present situation with antimicrobial susceptibility of N. gonorrhoeae in Russia and demonstrated the possibility of using hydrogel microarrays to control the spread of antibiotic resistance.

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“…Targeted gene sequencing based on the Ion AmpliSeq™ platform (Thermo Fischer Scientific, USA) or AmpliSeq for Illumina products can be used for accurate genotyping of antimicrobial resistance determinants. An alternative technology that has successfully been proven useful for simultaneous identification of multiple specific genomic targets is matrix oligonucleotide microarray analysis, which is currently widely used [11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Hydrogel Droplet Microarray for Genotyping Antimicrobial Resistance Determinants in Neisseria gonorrhoeae Isolates

Shaskolskiy¹,

Kandinov²,

Kravtsov³

et al. 2022

Prime Archives in Environmental Research

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Author Contributions: B.S. designed and directed the project, performed statistical analysis. I.K. performed experiments on amplification and microarray hybridization. D.K. analyzed DNA samples using microarrays. A.V. selected gel composition and performed Sanger sequencing. S.G. optimized microarray hybridization assay. M.F. realized hydrogel microarrays manufacturing. A.K. supervised N. gonorrhoeae sample collection. V.S. and D.D. conducted N. gonorrhoeae antimicrobial susceptibility testing, E.D. performed validation of the assay, wrote the manuscript. D.G. designed and supervised the project, wrote the manuscript. All authors have read and agreed to the published version of the manuscript.

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Prediction of ceftriaxone MIC inNeisseria gonorrhoeae usingDNA microarray technology and regression analysis

Cited by 10 publications

References 35 publications

In vitro Susceptibility to β-Lactam Antibiotics and Viability of Neisseria gonorrhoeae Strains Producing Plasmid-Mediated Broad- and Extended-Spectrum β-Lactamases

In vitro Susceptibility to β-Lactam Antibiotics and Viability of Neisseria gonorrhoeae Strains Producing Plasmid-Mediated Broad- and Extended-Spectrum β-Lactamases

Hydrogel Droplet Microarray for Genotyping Antimicrobial Resistance Determinants in Neisseria gonorrhoeae Isolates

Hydrogel Droplet Microarray for Genotyping Antimicrobial Resistance Determinants in Neisseria gonorrhoeae Isolates

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