Oligonucleotide Microchip for the Identification of Infectious Agents of Reproductive System with Simultaneous Analysis of Determinants of Resistance to Antimicrobial Substances

Leinsoo, Arvo; Шаскольский, Б Л; Dement'eva, E. I.; Gryadunov, Dmitry; Kubanov, Alexey А.; Chestkov, A. V.; Образцова, О. А.; Shpilevaya, M. V.; Deryabin, D. G.

doi:10.1007/s10517-017-3925-5

Cited by 9 publications

(10 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The microarray was previously developed for the identification of causative agents of human reproductive tract infections, including N . gonorrhoeae , and for the simultaneous detection of genetic markers of resistance to different antimicrobial drugs [36]. The microarray consisted of elements with immobilized oligonucleotides for the detection of different mutations and other determinants associated with resistance to beta-lactams: mutations in penA resulting in the insertion of Asp in the 345 position in PBP2 (insAsp345), mutations in ponA resulting in the amino acid substitution Leu421Pro in PBP1, the bla TEM plasmid and Met182Thr and Gly238Ser substitutions in the gene encoding beta-lactamase, mutations in porB leading to the amino acid changes Gly120Lys/Asp/Asn/Thr and Ala121/Asp/Asn/Gly/Ser in the porin protein, and deletion A (delA) and insertions T and TT (insT and insTT) in the promoter region of mtrR .…”

Section: Methodsmentioning

confidence: 99%

Resistance of Neisseria gonorrhoeae isolates to beta-lactam antibiotics (benzylpenicillin and ceftriaxone) in Russia, 2015–2017

et al. 2019

View full text Add to dashboard Cite

The goal of this work was to study the phenotypic susceptibility and resistance determinants of N . gonorrhoeae isolates to beta-lactam antimicrobials (benzylpenicillin and ceftriaxone). A total of 522 clinical isolates collected in Russia in 2015–2017 were analysed for susceptibility using the agar dilution method. DNA loci involved in antimicrobial resistance were identified using DNA microarray analysis and sequencing. Resistance to benzylpenicillin remained high, with 7.7% of isolates resistant (MIC pen > 1 mg/L) and 47.5% of isolates showing intermediate susceptibility (MIC pen = 0.12–1 mg/L). The most frequent resistance determinant (72.4% isolates) was the Asp345 insertion in penA , both as a single mutation and in combination with other mutations, particularly with the substitution Leu421Pro in ponA (39.0%). Mutations affecting the influx and efflux of drugs were also found, including amino acid substitutions in PorB (26.8% isolates) and delA in the promoter region of mtrR (22.8%). The accumulation of mutations in chromosomal genes ( penA , pon , porA , and mtrR ) led to a stepwise increase in MIC pen to values characteristic of intermediate resistance. The presence of bla TEM plasmids was found in 25 isolates (4.8%), resulting in a strong increase in resistance to penicillin (MIC pen > 16 mg/L) compared with the chromosomal mutations; 23 plasmids were of the African type with TEM-1 beta-lactamase, and two plasmids were of the Toronto/Rio type with TEM-135 beta-lactamase. Only three isolates were found with reduced susceptibility to ceftriaxone, with MIC cef = 0.12–0.25 mg/L. Sequencing of penA did not reveal mutations associated with resistance to third-generation cephalosporins, and the gene structure was non-mosaic. The majority of isolates (21 of 25) carrying the bla TEM plasmid also contained the conjugative plasmid with tetM (resistance to tetracyclines), consistent with previously reported data that the presence of the conjugative plasmid facilitates the transfer of other plasmids associated with antimicrobial resistance.

show abstract

Section: Methodsmentioning

confidence: 99%

Resistance of Neisseria gonorrhoeae isolates to beta-lactam antibiotics (benzylpenicillin and ceftriaxone) in Russia, 2015–2017

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Genetic determinants of antimicrobial resistance were identified using an oligonucleotide hydrogel-based low-density microarray as described previously [ 34 , 35 ]. The analyzed determinants associated with resistance to penicillin G [ 12 , 23 ] were as follows: mutations in the ponA gene resulting in the amino acid substitution Leu421Pro in penicillin-binding protein 1 (PBP1), mutations in the penA gene resulting in the insertion of an aspartate in the 345 position (insAsp345a) of PBP2, and the presence of bla TEM plasmids encoding β-lactamases capable of hydrolyzing amide bonds in penicillins.…”

Section: Methodsmentioning

confidence: 99%

Molecular Typing of Neisseria gonorrhoeae Clinical Isolates in Russia, 2018–2019: A Link Between penA Alleles and NG-MAST Types

et al. 2020

View full text Add to dashboard Cite

This work aimed to study penA gene polymorphisms in clinical isolates of Neisseria gonorrhoeae collected in Russia in 2018–2019 and the contribution of the penA allele type to susceptibility to β-lactam antibiotics. A total of 182 isolates were analyzed. penA allele types were determined by sequencing, and the minimum inhibitory concentrations (MICs) of benzylpenicillin and ceftriaxone were measured. The influence of genetic factors on MICs was evaluated by regression analysis. All isolates were susceptible to ceftriaxone, and 40.1% of isolates were susceptible to penicillin. Eleven penA allele types were identified. The mosaic type XXXIV penA allele and the Gly120Lys substitution in PorB made the greatest contributions to increasing the ceftriaxone MIC; the presence of the blaTEM plasmid, Gly120Asp, Ala121Gly/Asn substitutions in PorB, and the adenine deletion in the promoter region of the mtrR gene caused an increase in the penicillin MIC. Among 61 NG-MAST types identified, the most frequent were types 228, 807, 9486, 1993, and 6226. A link between penA alleles and Neisseria gonorrhoeae multi-antigen sequence typing (NG-MAST) types was established. Resistance to two groups of β-lactam antibiotics was associated with non-identical changes in penA alleles. To prevent the emergence of ceftriaxone resistance in Russia, NG-MAST genotyping must be supplemented with penA allele analysis.

show abstract

“…A microarray ( Fig. 5A ) and a procedure for its use have been developed to identify the DNAs of 12 different obligate and opportunistic microorganisms and simultaneously perform the differential analysis of 39 genetic determinants of resistance to β-lactam antibiotics, macrolides, aminoglycosides, tetracycline, spectinomycin, fluoroquinolones, and nitroimidazole [33].…”

Section: Analysis Of Specific Sequences Of Bacterial and Viral Genomesmentioning

confidence: 99%

The EIMB Hydrogel Microarray Technology: Thirty Years Later

Gryadunov¹,

Shaskolskiy²,

Наседкина³

et al. 2018

Acta Naturae

Self Cite

View full text Add to dashboard Cite

Biological microarrays (biochips) are analytical tools that can be used to implement complex integrative genomic and proteomic approaches to the solution of problems of personalized medicine (e.g., patient examination in order to reveal the disease long before the manifestation of clinical symptoms, assess the severity of pathological or infectious processes, and choose a rational treatment). The efficiency of biochips is predicated on their ability to perform multiple parallel specific reactions and to allow one to study the interactions of biopolymer molecules, such as DNA, proteins, glycans, etc. One of the pioneers of microarray technology was the Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences (EIMB), with its suggestion to immobilize molecular probes in the three-dimensional structure of a hydrophilic gel. Since the first experiments on sequencing by hybridization on oligonucleotide microarrays conducted some 30 years ago, the hydrogel microarrays designed at the EIMB have come a long and successful way from basic research to clinical laboratory diagnostics. This review discusses the key aspects of hydrogel microarray technology and a number of state-ofthe-art approaches for a multiplex analysis of DNA and the protein biomarkers of socially significant diseases, including the molecular genetic, immunological, and epidemiological aspects of pathogenesis. KEYWORDS hydrogel microarrays, nucleic acid hybridization, multiplex immunochemical assay, antimicrobial drug resistance, genotyping, tumor markers. ABBREVIATIONS NAs -nucleic acids; NGS -next-generation sequencing; FDA -Food and Drug Administration (USA); MTB -Mycobacterium tuberculosis, causative agent of tuberculosis; NTM -non-tuberculous mycobacteria, causative agents of mycobacteriosis; HCV -hepatitis C virus; RMP -rifampin; INH -isoniazid; EMBethambutol; MDR -multidrug resistance; XDR -extensive drug resistance; RTIs -reproductive tract infections; AMD -antimicrobial drugs; CRC -colorectal carcinoma; CEA -carcinoembryonic antigen; CA -carbohydrate antigen. REVIEWS VOL. 10 № 4 (39) 2018 | ACTA NATURAE | 5

show abstract

Oligonucleotide Microchip for the Identification of Infectious Agents of Reproductive System with Simultaneous Analysis of Determinants of Resistance to Antimicrobial Substances

Cited by 9 publications

References 10 publications

Resistance of Neisseria gonorrhoeae isolates to beta-lactam antibiotics (benzylpenicillin and ceftriaxone) in Russia, 2015–2017

Resistance of Neisseria gonorrhoeae isolates to beta-lactam antibiotics (benzylpenicillin and ceftriaxone) in Russia, 2015–2017

Molecular Typing of Neisseria gonorrhoeae Clinical Isolates in Russia, 2018–2019: A Link Between penA Alleles and NG-MAST Types

The EIMB Hydrogel Microarray Technology: Thirty Years Later

Contact Info

Product

Resources

About