A Method for Processing Digital Images of Colorimetric Biochipes for Quantitative Determination for Bacterial Antibiotic Resistance

2021 ◽  
Vol 37 (5) ◽  
pp. 123-131
Author(s):  
G.V. Presnova ◽  
V.G. Grigorenko ◽  
M.M. Ulyashova ◽  
М.Yu. Rubtsova
Keyword(s):  
Antibiotic Resistance ◽  
Nucleic Acids ◽  
Quantitative Determination ◽  
Resistance Genes ◽  
Molecular Genetic ◽  
Antibiotic Resistance Genes ◽  
Molecular Genetic Analysis ◽  
Beta Lactamases ◽  
The Government

Abstract-Molecular genetic analysis methods based on the technology of colorimetric biochip have shown their effectiveness in identifying antibiotic resistance genes in bacteria. For the quantitative determination of nucleic acids, a comparative study of methods for converting digital color images of biochips into monochrome black-and-white versions using RGB and CMYK color models has been carried out. A 19-mer single-stranded oligonucleotide and two model mRNAs corresponding to the genes of two types of clinically relevant beta-lactamases (CTX-M and NDM) were studied as objects. The widest range of staining intensity and the best analytical characteristics for the determination of all types of studied nucleic acids were obtained using the red channel of the RGB color model. The detection limits were 0.10 ± 0.02 pmol/μl for the 19-mer oligonucleotide, and 3.0 ± 0.2 amol/μl and 8.0 ± 0.6 amol/μl for mRNA of beta-lactamases CTX-M-116 and NDM-1, respectively. The developed method can be used for the quantitative determination of expressing antibiotic resistance genes in bacteria with multiple resistance to antimicrobial drugs. Key words: colorimetric biochips, hybridization analysis, DNA, mRNA, antibiotic resistance, beta-lactamases The work was supported by the Government Program of the Lomonosov Moscow State University (АААА-А21-121011290089-4).

Determination of antibiotic resistance genes in a WWTP-impacted river in surface water, sediment, and biofilm: Influence of seasonality and water quality

2021 ◽  
pp. 144526
Author(s):  
Gabriela Reichert ◽  
Stephan Hilgert ◽  
Johannes Alexander ◽  
Júlio César Rodrigues de Azevedo ◽  
Tobias Morck ◽  
...  
Keyword(s):  
Water Quality ◽  
Antibiotic Resistance ◽  
Surface Water ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes

scholarly journals Analysis of Class 1 Integrons and Antibiotic Resistance Genes in Pseudomonas aeruginosa Strains from Benin City, Nigeria

2020 ◽  
Vol 24 (4) ◽  
pp. 633-637
Author(s):  
B.O. Isichei-Ukah ◽  
O.I. Enabulele
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Environmental Isolates ◽  
Significance Level ◽  
Beta Lactamases ◽  
Class 1 Integrons ◽  
Benin City ◽  
Class 1

The presence of integrons and antibiotic resistance genes in the genome of Pseudomonas aeruginosa pose a serious problem in the treatment and control of infections caused by this pathogen in hospitals. This study was carried to analyse the presence of class 1 integrons and some antibiotic resistance genes on selected clinical and environmental strains of Pseudomonas aeruginosa. A total of 120 strains were employed for this study.The strains were confirmed using molecular method and species-specific primers targeting the 16S ribosomal ribonucleic acid (rRNA). Polymerase chain reaction (PCR) was used to detect the presence of class 1 integrons and resistance genes using appropriate primers and conditions. The strains were analysed for the presence of the following antibiotic resistance genes - aadA, blaPSE, blaAMPC, blaIMP and tetC encoding  aminoglycosides, betalactamases, metallo-beta-lactamases (MBL) and tetracylines resistance respectively. On screening the isolates for the presence of class 1 integrons, 50/60 (83.3 %) clinical isolates and 46/60 (76.7 %) environmental isolates showed positive results (P > 0.05). In both clinical and environmental isolates, the highest occurring resistance genes were blaAMPC and tetC (encoding beta-lactamases and tetracylines respectively), while the least was observed in blaIMP (encoding metallo-beta-lactamases). In comparison, there was high significance difference (at P<0.01 significance level) in the resistance gene blaPSE between the clinical and environmental strains. The high prevalence of these resistance genes is a great threat in the treatment of Pseudomonas infections. Keywords: Pseudomonas aeruginosa, Resistance genes, Integrons, Beta-lactamases.

GENERAL PRINCIPLES OF ANTIBIOTIC RESISTANCE EVOLUTION IN BACTERIA (REVIEW OF LITERATURE)

2020 ◽  
Vol 65 (6) ◽  
pp. 387-393
Author(s):  
N. V. Davidovich ◽  
Natalya Nilolaevna Kukalevskaya ◽  
E. N. Bashilova ◽  
T. A. Bazhukova
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Molecular Genetic ◽  
Antibiotic Resistance Genes ◽  
Human Pathogens ◽  
Intrinsic Resistance ◽  
Resistance Evolution ◽  
Genes Encoding ◽  
The Impact

Currently, the impact of antibiotic resistance on human health is a worldwide problem and its study is of great interest from a molecular genetic, environmental and clinical view-point. This review summarizes the latest data about antibiotic resistance, the classification of microorganisms as sensitive and resistant to the action of antibiotics, reveals the concept of minimum inhibitory concentration from modern positions. The resistance of microorganisms to antibacterial agents can be intrinsic and acquired, as well as being one of the examples of evolution that are currently available for study. Modern methods of whole-genome sequencing and complex databases of nucleotide-tagged libraries give an idea of the multifaceted nature of the mechanisms of intrinsic resistance to antibiotics and are able to provide information on genes encoding metabolic enzymes and proteins that regulate the basic processes of the physiology of bacteria. The article describes the main ways of spreading the resistance of microorganisms, reflects the concepts of “founder effect” and the fitness cost of bacteria, which underlie the emergence and evolution of antibiotic resistance. It is shown that the origin of antibiotic resistance genes that human pathogens currently possess can be traced by studying the surrounding not only clinical, but also non-clinical (ecological) habitats. As well as microorganisms of the surrounding ecosystems are the donors of resistance genes in horizontal gene transfer.

scholarly journals Evolution of Susceptibility of Strains of Candida Fluconazole and Determination of Antibiotic Resistance Genes

2018 ◽  
Vol 5 (2) ◽  
Author(s):  
Khadije Rezaie Keikhaie ◽  
Fatemeh Moshtaghi ◽  
Forough Forghani ◽  
Maryam Sheykhzade Asadi ◽  
Samira Seyed Nejad ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes

Concentration of facultative pathogenic bacteria and antibiotic resistance genes during sewage treatment and in receiving rivers

2016 ◽  
Vol 74 (8) ◽  
pp. 1753-1763 ◽  
Author(s):  
Stefanie Heß ◽  
Frauke Lüddeke ◽  
Claudia Gallert
Keyword(s):  
Antibiotic Resistance ◽  
River Water ◽  
Resistance Genes ◽  
Pathogenic Bacteria ◽  
Quantitative Polymerase Chain Reaction ◽  
Molecular Genetic ◽  
Sewage Treatment ◽  
Antibiotic Resistance Genes ◽  
Qpcr Data ◽  
Total Bacteria

Whereas the hygienic condition of drinking and bathing water by law must be monitored by culture-based methods, for quantification of microbes and antibiotic resistance in soil or the aquatic environment, often molecular genetic assays are used. For comparison of both methods, knowledge of their correlation is necessary. Therefore the population of total bacteria, Escherichia coli, enterococci and staphylococci during sewage treatment and in receiving river water was compared by agar plating and quantitative polymerase chain reaction (qPCR) assays. In parallel, all samples were investigated for clinically relevant antibiotic resistance genes. Whereas plating and qPCR data for total bacteria correlated well in sewage after primary treatment, qPCR data of river water indicated higher cell numbers for E. coli. It is unknown if these cells are ‘only’ not growing under standard conditions or if they are dead. Corresponding to the amount of non-culturable cells, the ‘breakpoints’ for monitoring water quality should be adapted. The abundances of clinically relevant antibiotic resistance genes in river water were in the same order of magnitude or even higher than in treated sewage. For estimation of the health risk it is important to investigate which species carry respective genes and whether these genes are disseminated via gene transfer.

scholarly journals Study on Bacteria Isolates and Antimicrobial Resistance in Wildlife in Sicily, Southern Italy

2021 ◽  
Vol 9 (1) ◽  
pp. 203
Author(s):  
Delia Gambino ◽  
Domenico Vicari ◽  
Maria Vitale ◽  
Giorgia Schirò ◽  
Francesco Mira ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Wild Animals ◽  
Residential Areas ◽  
Bacterial Strains ◽  
Urban Centers ◽  
Biochemical Tests ◽  
Zoonotic Bacteria

Wild environments and wildlife can be reservoirs of pathogens and antibiotic resistance. Various studies have reported the presence of zoonotic bacteria, resistant strains, and genetic elements that determine antibiotic resistance in wild animals, especially near urban centers or agricultural and zootechnical activities. The purpose of this study was the analysis, by cultural and molecular methods, of bacteria isolated from wild animals in Sicily, Italy, regarding their susceptibility profile to antibiotics and the presence of antibiotic resistance genes. Bacteriological analyses were conducted on 368 wild animals, leading to the isolation of 222 bacterial strains identified by biochemical tests and 16S rRNA sequencing. The most isolated species was Escherichia coli, followed by Clostridium perfringens and Citrobacter freundii. Antibiograms and the determination of resistance genes showed a reduced spread of bacteria carrying antibiotic resistance among wild animals in Sicily. However, since several wild animals are becoming increasingly close to residential areas, it is important to monitor their health status and to perform microbiological analyses following a One Health approach.

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