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Detection of the effect of synthetic siRNA on efflux pump MexA gene expression and antibiotic resistance in clinical isolates of Pseudomonas aeruginosa
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Introduction and Aim: Pseudomonas aeruginosa is a nosocomial infection with an ability to develop high levels of antibiotic resistance. The efflux pump system is one of the mechanisms that is linked to multidrug resistance in P. aeruginosa. In this study, we employed siRNA loaded on gold nanoparticles against the MexA efflux pump gene to decrease the MexA gene expression in P. aeruginosa and estimated antibiotic resistance after gene silencing.   Materials and Methods: This study examined four strains of P. aeruginosa isolated from patients in various hospitals in Baghdad. Bacteria isolated were identified by biochemical tests and Vitek compact 2 system.  Single-stranded siRNA (33bp) designed in this study was loaded onto gold nanoparticles (AuNPls). Detection of the MexA gene was carried out by conventional PCR technique. The expression of MexA gene was examined by qRT-PCR in order to determine if the siRNA have impacted on MexA gene expression and on the antibiotic resistance in aeruginosa   Results: This study showed that the mRNA expression level of the MexA gene exhibited a decrease in fold change CT -2   in P. aeruginosa (isolates numbers 67, 66,49, and PDR(5p)) when examined in vitro. The specific fold change values observed were (0.202, 0.040, 0.063, and 0.163) respectively. The resistance percentages of antibiotics tested was observed to increase after MexA gene silencing.   Conclusion: Targeting the MexA gene with synthetic siRNA may be a unique approach to diminish P. aeruginosa resistance to antibiotics. However, many unexpected consequences may occur when utilizing any genetic manipulation in bacteria.

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Publication Date
Fri Jun 23 2023
Journal Name
Journal Of Communicable Diseases
Downregulation of the Gene Expression of Efflux Pump Gene (acrA) and its Regulator (ramA) in Klebsiella pneumoniae by Quaternary Ammonium Compounds
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Introduction: Biocides are commonly used for disinfection in a variety of contexts. They are generally used to avoid infection by controlling biofilm on medical equipment. However, the literature lacks information on the effect of biocide on efflux pump gene expression. Objective: To determine the influence of biocide on biofilm development and efflux pump acrA and ramA gene expression. Methodology: The microtiter plate method was used to identify biofilm development in 80 isolates of K. pneumoniae. The minimal inhibitory concentrations (MIC) of three biocides (quaternary ammonium compound (QAC), chlorohexidine digluconate, and chloroxylenol) were estimated. The effect of QAC on the intensity and viability of biofilms was investigated as we

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Publication Date
Sat Sep 30 2023
Journal Name
Iraqi Journal Of Science
Study on tssC1 Gene Mediating Biofilm Antibiotics Resistance of Pseudomonas aeruginosa
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P. aeruginosa is a famous bacterium that causes several diseases and has a high ability to be a multidrug resistant organism that is linked with the formation of biofilm. This study aimed to investigate tssC1 gene role in the resistance of different antibiotics in the presence of biofilm. We constructed biofilm for the isolates under the study and showed the effect of different antibiotics on biofilm formation and maturation. The presence of the gene was detected through achieving PCR reaction. Finally, tssC1 gene variation was determined through sequencing and aligning the sequencing products. The results showed that most of the isolates (80%) formed biofilm that played a role in the resistance of different antibiotics which could

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Publication Date
Thu Feb 27 2020
Journal Name
Iraqi Journal Of Science
Gene Expression of pelA and pslA in Pseudomonas Aeruginosa under Gentamicin Stress
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     Pseudomonas aeruginosa produces an extracellular biofilm matrix that consists of nucleic acids, exopolysaccharides, lipid vesicles, and proteins. Alginate, Psl and Pel are three exopolysaccharides that constitute the main components in biofilm matrix, with many biological functions attributed to them, especially concerning the protection of the bacterial cell from antimicrobial agents and immune responses. A total of 25 gentamicin-resistant P. aeruginosa selected isolates were enrolled in this study. Biofilm development was observed in 96% of the isolates. In addition, the present results clarified the presence of pelA and pslA in all the studied isolates. The expression of these genes was very low. Even though all biof

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Publication Date
Wed Aug 31 2022
Journal Name
Al-kindy College Medical Journal
Antimicrobial Activity of Lepidium Sativum against Multi drug resistant and sensitive Pseudomonas aeruginosa from clinical isolates, Khartoum State, Sudan: Lepidium Sativum against Clinical isolates of Pseudomonas aeruginosa
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Background: L. sativum, are traditionally used for the treatment of various diseases and thought to have medicinal value. Isolates from many part of the world is now multidrug resistant. Therefore, there is an urgent need to look for and test an alternative herbal drug.

Objective: The present study aimed to evaluate the antibacterial activity of L. Sativum seed extract against multi drug resistant (MDR) and sensitive Pseudomonas aeruginosa clinical isolates.

Subjects and Methods: An ethanolic and aqueous stock extracts were prepared from L.  sativum seed plant then serial dilutions were prepared and the obtained concentrations (50, 25, 12.5 and 6.2 mg/ml) were tested against 30 multidrug-resistan

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Publication Date
Thu Oct 01 2020
Journal Name
Indian Journal Of Forensic Medicine & Toxicology
Study the Ability of Pseudomonas Aeruginosa Isolated from Different Clinical Cases to Biofilm Formation and Detection of Algd Gene.
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98 samples were collected from various clinical sources included (Burns, wounds, urines, sputums, blood) From the city of Baghdad, After performing the biochemical and microscopic examination, 52 isolates were obtained for Pseudomonas aeruginosa, 17 (32.7%) isolates from burn infection, 12 (23%) isolates from Wound infection 11 (21.2%) isolates from urine infection, 7 (13.5%) isolates of sputum and 5 (9.6%) isolates from blood. Bacteria susceptibility to form biofilm has been detectedby microtiter plate method, The results showed that 80% of the bacterial isolates were produced the biofilm with different proportions, alg D gene (alginate production) has been detected by polymerase chain reaction (PCR) Which plays an essential role in the fo

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Publication Date
Tue Jan 03 2017
Journal Name
World Journal Of Experimental Biosciences
Effect of Soil Extracts on Ability of Clinical Isolates of Pseudomonas aeruginosa and Staphylococcus aureus to Produce Biofilm in vitro
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In present study the effect of soil extracts of different types of soil on ability of two clinical isolates, Pseudomonas aeruginosa and Staphylococcus aureus to form biofilm. The extract of soil was done by using sterile phosphate buffer saline and analyzed by Fourier Transform Infrared Spectroscopic (FTIR). Spectrophotometric method was used to check ability of the studied isolated bacteria to form biofilm on polystyrene microtiter plates. The data of FTIR showed very little difference was observed among extracts of three types of soil (soil contaminated with hydrocarbons; garden soil collected from gardens of al-jadrea, Baghdad and containers soil), but the highest difference was observed in the extract obtained from peat moss clay soil.

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Publication Date
Thu Dec 15 2022
Journal Name
Bionatura
Antibiogram of Eucalyptus and Sesame seed oil against clinical isolates of Pseudomonas aeruginosa
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Publication Date
Tue Jan 01 2019
Journal Name
Iraqi Journal Of Agricultural Sciences
Cloning and expression of a lipase gene from Pseudomonas aeruginosa into E.coli
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Fifteen local isolates of Pseudomonas were obtained from several sources such as soil, water and some high-fat foods (Meat, olives, coconuts, etc.). The ability of isolates to produce lipase was measured by the size of clear zone on Tween 20 solid medium and by measuring the enzymatic activity and specific activity. Isolate M3 (as named in this study) was found to be the most efficient for the production of the lipase with enzymatic activity reached 56.6 U/ml and specific activity of 305.94 U/mg. This isolate was identified through genetic analysis of the 16S rRNA gene. and it was shown that the isolate M3 belongs to Pseudomonas aeruginosa with 99% similarity. The DNA of isolate M3 was extracted and lipase gene was amplified through PCR tec

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Scopus
Publication Date
Sun Jul 31 2022
Journal Name
Iraqi Journal Of Science
Gentamicin Variably Affects amrZ and rhl gene Expression in Swarmer Cells of Pseudomonas aeruginosa
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       Swarming is one of the most important virulence factors used by bacteria to invade new sites. This study aimed to test the effects of gentamicin on swarming motility of Pseudomonas aeruginosa, both phenotypically and molecularly. The present results revealed that 11/25 isolates had gentamicin MIC of 1024 µg/ml.  However, gentamicin at sub-minimal inhibitory concentration significantly (P< 0.05) reduced the diameter of swarming in all P. aeruginosa isolates. Noticeably the mean and median swarming diameter before treatment with gentamicin 5.557 and 5.816 cm respectively had significantly (P < 0.001) reduced to 0.871 and 0.766 cm respectively. At the molecular level, amrZ (a global regulator of multiple genes) and

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Publication Date
Thu Mar 09 2017
Journal Name
Ibn Al-haitham Journal For Pure And Applied Sciences
DNA Sequences of LasB Gene in Pseudomonas aeruginosa Isolated from Some Clinical Cases
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 Out of 120 isolates from different clinical cases, only 75 were found and confirmed that they belong to the Pseudomonas aeruginosa bacteria. The result revealed that the LasB virulent gene was present in 63 isolates with 63% percentage. The gel electrophoresis showed that the molecular weight of LasB gene was 300 bp. DNA sequences of LasB gene was done, and the results showed the presence of some gene mutations like substitution, addition and deletion with 97% identity with the Refseq gene. From the other side, the results of identities of translated nucleotides sequence with the original sequence of amino acids revealed that there are no effects of gene mutations on translation of the product protein. 

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