scholarly journals The impact of Grammosciadium platycarpum Boiss. & Hausskn. extract on oqxA efflux pump gene expression in antibiotic resistant clinical isolates of Klebsiella pneumoniae using real time PCR

2020 ◽  
Vol 19 (75) ◽  
pp. 291-304
Author(s):  
Faezeh Mohammadpour Bishak ◽  
Fatemeh Ashrafi ◽  
Soheila Moradi Bidhendi ◽  
Amir Mirzaie ◽  
Hassan Noorbazargan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Klebsiella Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
Efflux Pump ◽  
Clinical Isolates ◽  
Antibiotic Resistant ◽  
The Impact

scholarly journals SENSITIVITY OF KLEBSIELLA PNEUMONIAE CLINICAL ISOLATES WITH VARIOUS LEVELS OF ANTIBIOTIC RESISTANCE TO BACTERIORAGE PREPARATIONS

2018 ◽  
pp. 56-62
Author(s):  
D. V. Tapalsky ◽  
A. I. Kozlova
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
High Activity ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Isolates ◽  
Antibiotic Resistant ◽  
Medical Institutions ◽  
Different Levels

Objective : to assess sensitivity of K. pneumoniae clinical isolates with different levels of antibiotic resistance to commercial bacteriophage preparations. Material and methods . We have performed re-identification and determination of the sensitivity to antibiotics and bacteriophage preparations of 109 K. pneumoniae clinical isolates isolated from patients hospitalized in medical institutions of three regions of Belarus. The presence of carbapenemase genes has been deteсted by real-time PCR. Results . The study has shown awidespread prevalence of extreme antibiotic resistance among K. pneumoniae associated with the production of NDM and OXA-48 carbapenemases and has found an insufficient lytic activity of bacteriophage preparations against K. pneumoniae strains. The preparation «Sextafag», which lysed with a high activity of 28.4% of Klebsiella isolates possessed the widest spectrum of activity. Conclusion . Bacteriophage preparations can be considered as a possible alternative to antibiotics in the treatment of infections caused by extremely antibiotic-resistant K. pneumoniae isolates.It is necessary to supplement the composition of commercial preparations with phage strains that are active against Klebsiella isolates producing carbapenemases.

scholarly journals Selection and validation of reference genes for gene expression studies in Klebsiella pneumoniae using Reverse Transcription Quantitative real-time PCR

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Ana Érika Inácio Gomes ◽  
Leonardo Prado Stuchi ◽  
Nathália Maria Gonçalves Siqueira ◽  
João Batista Henrique ◽  
Renato Vicentini ◽  
...  
Keyword(s):  
Gene Expression ◽  
Klebsiella Pneumoniae ◽  
Real Time ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Quantitative Real Time Pcr ◽  
Expression Studies ◽  
Gene Expression Studies

scholarly journals Use of Real-Time PCR and Fluorimetry for Rapid Detection of Rifampin and Isoniazid Resistance-Associated Mutations in Mycobacterium tuberculosis

2000 ◽  
Vol 38 (9) ◽  
pp. 3194-3199 ◽  
Author(s):  
Maria J. Torres ◽  
Antonio Criado ◽  
Jose C. Palomares ◽  
Javier Aznar
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Antimicrobial Agents ◽  
Dna Amplification ◽  
Single Strand Conformation Polymorphism ◽  
Clinical Isolates ◽  
Polymorphism Analysis ◽  
Antibiotic Resistant ◽  
Hybridization Probes

Very fast amplification of DNA in small volumes can be continuously monitored with a rapid cycler that incorporates fluorimetric detection. Primers were designed to amplify a 157-bp fragment of therpoB gene spanning codons 526 and 531 and a 209-bp fragment of the katG gene spanning codon 315 of Mycobacterium tuberculosis. Most mutations associated with resistance to rifampin (RMP) and isoniazid (INH) in clinical isolates occur in these codons. Two pairs of hybridization probes were synthesized; one in each pair was 3′ labeled with fluorescein and hybridized upstream of the codon with the mutation; the other two probes were 5′ labeled with LightCycler-Red 640. Each pair of probes recognized adjacent sequences in the amplicon. After DNA amplification was finished by using a LightCycler, the temperature at which the Red 640 probe melted from the product was determined in a 3-min melt program. Twenty M. tuberculosis clinical isolates susceptible to streptomycin, INH, RMP, and ethambutol and 36 antibiotic-resistant clinical M. tuberculosis isolates (16 resistant to RMP, 16 to INH, and 4 to both antimicrobial agents) were amplified, and the presence of mutations was determined using single-strand conformation polymorphism analysis, the LiQor automated sequencer, and the LightCycler system. Concordant results were obtained in all cases. Within 30 min, the LightCycler method correctly genotyped all the strains without the need of any post-PCR sample manipulation. Overall, this pilot study demonstrated that real-time PCR coupled to fluorescence detection is the fastest available method for the detection of RMP and INH resistance-associated mutations in M. tuberculosis clinical isolates.

scholarly journals Real-Time PCR and Statistical Analyses of acrAB and ramA Expression in Clinical Isolates of Klebsiella pneumoniae

2008 ◽  
Vol 52 (9) ◽  
pp. 3430-3432 ◽  
Author(s):  
Alexey Ruzin ◽  
Frederick W. Immermann ◽  
Patricia A. Bradford
Keyword(s):  
Klebsiella Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
Statistical Analyses ◽  
Clinical Isolates ◽  
Susceptibility Breakpoint

ABSTRACT Clinical isolates of Klebsiella pneumoniae were tested for a correlation between tigecycline MIC and expression of ramA by using real-time PCR. At MICs of 4 and 8 μg/ml, the expression of ramA was statistically significantly different from MICs of 2 μg/ml or less, supporting the tigecycline susceptibility breakpoint of ≤2 μg/ml for K. pneumoniae.

scholarly journals Multiplex Real-Time PCR for Detection of an Epidemic KPC-Producing Klebsiella pneumoniae ST258 Clone

2012 ◽  
Vol 56 (6) ◽  
pp. 3444-3447 ◽  
Author(s):  
Liang Chen ◽  
Kalyan D. Chavda ◽  
José R. Mediavilla ◽  
Yanan Zhao ◽  
Henry S. Fraimow ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Isolates ◽  
Pcr Assay ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Sequence Types ◽  
Single Reaction

ABSTRACTWe describe a multiplex real-time PCR assay capable of identifying both the epidemicKlebsiella pneumoniaeST258 clone andblaKPCcarbapenemase genes in a single reaction. The assay displayed excellent sensitivity (100%) and specificity (100%) for identification of ST258 clone andblaKPCin a collection of 75K. pneumoniaeisolates comprising 41 sequence types. Our results suggest that this assay is an effective tool for surveillance of this clone among carbapenem-resistantK. pneumoniaeclinical isolates.

scholarly journals Antibiotic Resistant, Virulence-associated Genes, Biofilm and Efflux Pump Gene Expression and Molecular Typing of Klebsiella Pneumoniae Strains Recovered from Clinical Samples

2020 ◽  
Author(s):  
Amir Mirzaie ◽  
Reza Ranjbar
Keyword(s):  
Gene Expression ◽  
Klebsiella Pneumoniae ◽  
Molecular Typing ◽  
Efflux Pump ◽  
Clinical Samples ◽  
Public Health Issue ◽  
Pcr Analysis ◽  
Antibiotic Resistant ◽  
Microbiological Methods ◽  
High Level

Abstract BackgroundMultidrug-resistant (MDR) Klebsiella pneumoniae strains are one of the most important life-threatening nosocomial pathogens. In the current study, antibiotic resistant, virulence-associated genes, gene expression of efflux pumps and biofilm genes as well as molecular typing of K. pneumoniae strains were investigated. A total of 505 clinical specimens were collected from hospitalized patients and K. pneumoniae strains were isolated by standard microbiological methods. Antibiotic resistant profile, prevalence of virulence associated genes, biofilm and efflux pump genes were investigated. The gene expression analysis of biofilm and efflux pump genes were analused quantitative Real Time PCR. Moreover, molecular typing of K. pneumoniae strains using Repetitive element sequence-based PCR (rep-PCR) technique was also carried out. ResultsOne hundred K. pneumoniae strains out of 500 clinical samples were isolated and the highest prevalence of resistance was observed against ciprofloxacin (75%), Trimethoprim-sulfamethoxazole (73%) and Nitrofurantoin (38%). Virulence associated genes including entB, traT and rmpA were found in 80%, 62% and 48%, respectively. Gene prevalence for biofilm association gene including mrkA, fimH and mrkD were 42% for all genes. The AcrAB, TolC and mdtK efflux pump genes were observed in 41%, 33% and 26%, respectively. In addition, most MDR strains formed biofilm, as well as, AcrAB efflux pump and mrkA biofilm gene expression was up-regulated in MDR K. pneumoniae strains and a significant statistically association was also observed between MDR strains and high expression of efflux pump and biofilm genes. In addition, the K. pneumoniae strains differentiated into 11 different genetic clusters by rep-PCR analysis. ConclusionsHigh prevalence of resistance, presence of diver’s virulence factors and high level of efflux pump and biofilm gene expression in diverse clones of K. pneumoniae strains pose an important public health issue.

scholarly journals Expression of Multidrug Efflux Pump GenesacrAB-tolC,mdfA, andnorEinEscherichia coliClinical Isolates as a Function of Fluoroquinolone and Multidrug Resistance

2010 ◽  
Vol 55 (2) ◽  
pp. 921-924 ◽  
Author(s):  
Michelle C. Swick ◽  
Sonia K. Morgan-Linnell ◽  
Kimberly M. Carlson ◽  
Lynn Zechiedrich
Keyword(s):  
Escherichia Coli ◽  
Multidrug Resistance ◽  
Real Time ◽  
Quantitative Study ◽  
Real Time Pcr ◽  
Efflux Pump ◽  
Clinical Isolates ◽  
Strongly Correlated ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump

ABSTRACTIn a single quantitative study, we measuredacrA,acrB,tolC,mdfA, andnorEexpression inEscherichia coliclinical isolates by using real-time PCR.acrAandacrBoverexpression strongly correlated with fluoroquinolone and multidrug resistance;tolC,mdfA, andnorEexpression did not. The order of abundance of efflux pump transcripts in all fluoroquinolone-susceptible isolates wastolC(highest), thenacrAandacrB, and thenmdfAandnorE.Our findings suggestacrABoverexpression is an indicator of multidrug resistance.

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