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Developing multiplex PCR for the rapid and simultaneous detection of Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella pneumoniae associated with sheep respiratory tract infections
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Respiratory tract infections in sheep are among the important health problems that affect all sheep ages around the world. Nine bacterial isolates obtained from sheep with respiratory tract infections were selected to be used in the current study. The isolates included 3 Staphylococcus aureus, 4 Klebsiella pneumoniae, and 2 Pseudomonas aeruginosa. Following the primers design by the Primer3Plus software tool and optimization of the conventional polymerase chain reaction (PCR), the primers were validated for their use in the multiplex PCR experiments. The MFEprimer program was used to check the suitability of the primer set combinations for multiplex PCR. The MFEprimer software was successful in designing the multiplex-PCR experiments and determining the optimal primer set combinations. Multiplex PCR was able to amplify specific DNA sequences of one, two or three target genes of these mixed microorganisms in the same PCR reaction tube. This technique efficiently detected combinations of two organisms, either S. aureus with K. pneumoniae, S. aureus with P. aeruginosa or K. pneumoniae with P. aeruginosa. Moreover, multiplex PCR was also able to detect the presence of the three organisms together in the same reaction tube. To conclude, this study confirmed multiplex-PCR as a specific, sensi- tive, rapid, accurate, and cost-effective molecular diagnostic method for identification and differentiation of three clinically important bacteria associated with sheep respiratory tract infections, including S. aureus, P. aeruginosa, and K. pneumoniae. This can efficiently support control and treatment of such diseases and would increase the economy of the animals’ owners and wellbeing of the animals.

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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
Wed Mar 29 2017
Journal Name
Iraqi Journal Of Pharmaceutical Sciences ( P-issn 1683 - 3597 E-issn 2521 - 3512)
The Effects of Aloe vera Gel, Sesame Oil and Camphor Oil on Pseudomonas aeruginosa Isolated from Burnt Patients
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Three isolates of  P. aeruginosa were isolated from burnt patients. The ability of these isolates for adhesion and formation of slime layer were tested, the result showed that all isolates were able to adherence on the smooth surface. The sensitivity of  P. aeruginosa isolates for antibiotics were tested , all isolates were sensitive  to Gentamycin, Piperacillin and Amikacin Ciprofloxacin, and  resist to Tetracyclin, Amoxicillin, Cephalexine , Ceftriaxone. Ciprofloxacin and Amikacin were found effective against P. aeruginosa isolates with MIC values of 3.8 μg/ ml for  Ciprofloxacin  and 0.244 μg/ ml for Amikacin The antibacterial effect of Different concentrations of Aloe

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Publication Date
Fri Jun 30 2023
Journal Name
Biomedicin
Antibacterial action of AgNPs produced from different isolates Gram positive and Gram-negative bacteria on biofilm of Klebsiella pneumoniae isolated from the RTI
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Silver nanoparticles synthesized by different species

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Publication Date
Sat Sep 03 2022
Journal Name
International Journal Of Health Sciences
Using Elisa and nested PCR for detection of the toxoplasmosis in milk and the influence of infection and some factors on milk composition in the Iraqi local and Shami goats
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This study concluded detection of Toxoplasma gondii in milk, immunologically by using Elisa and nested PCR)nPCR (based on B1 gene, also to investigate the effect of toxoplasmosis, parity, breed and flock on some milk composition in the Iraqi local and Shami goats in the middle of Iraq. A total of 80 milk samples of the lactating goats were collected. Results of this study showed the prevalence of Toxoplasmosis was 21.25% and 28.75% by Elisa and nPCR respectively without significant differences. The sensitivity of Elisa was a low (30.43%) whereas the specificity was a high (82.45%). The degree of agreement estimated by Kappa coefficient revealed a slight agreement (0.14) between two methods. The results indicated that goats infected

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Publication Date
Tue May 01 2018
Journal Name
Journal Of Physics: Conference Series
The study of Bacteriocin of Pseudomonas fluorescens and Citrus limon effects against Propionibacterium acnes and Staphylococcus epidermidis in acne patients
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Research was carried out antibacterial of (Citrus limon) juice on Acnevulgaris. Samples were obtained from individuals having (Pimples) by swabbing their faces. Substances natural from plants are promising to treat disease cause Acnevulgaris, the study in vitro biological activity of the juice, as well as bacterocin cultivated and fruits was investigated on two strain bacteria (Propionibacterium acnes, Staphylococcus epidermidis). The new antimicrobial (bacteriocin and Citrus juice) is ongoing search. This study used s juice at different concentrations at (20%, 30%, 40%, 60%, 80% and 100%). The Bacteriocin produced from local P. fluorescens isolates from wound infection and majority of isolates were found to produce crude Bacteriocin were (

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Publication Date
Fri Jan 01 2021
Journal Name
Wiadomości Lekarskie
EVALUATION THE EFFECT OF PRODIGIOSIN ON ALGD EXPRESSION IN CLINICAL ISOLATES OF PSEUDOMONAS AERUGINOSA
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The aim of this study is to investigate the role of prodigiosin on P. aeruginosa' s biofilm genes involved in the pathogenicity and persistency of the bacteria; Materials and methods: Gram negative bacterial isolates were taken from burn and wounds specimen obtained from some of Baghdad hospitals. Forty six isolates were identified as Pseudomonas aeruginosa and four isolates as Serratia marcescens by using biochemical tests and VITEK 2 compact system. Susceptibility test was performed for all P. aeruginosa isolates, the results showed that 100% were resistant to Amikacin and 98% were sensitive to Meropenem. Resistant isolates were tested for biofilm formation; the strong and moderate isolates (17) were detected by PCR for AlgD gene

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Publication Date
Mon Aug 29 2016
Journal Name
World Journal Of Experimental Biosciences
Effect of sub-inhibitory and inhibitory concentrations of some antibiotics and rosemary essential oil (Rosmarinus officinalis L.) on biofilm formation of Klebsiella pneumoniae
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Klebsiella pneumoniae have an ability to form biofilm as one of strategies to persist and overcome host defenses. The study aims to evaluate the effectiveness of rosemary essential oil alone and in combination with some antibiotics against biofilm of K. pneumoniae isolated from urine. The antibiotics resistance pattern by disc diffusion method and minimal inhibitory concentration (MIC) of gentamicin, ciprofloxacin, amoxicillin, trimethoprim/ sulfame- thoxazole, cefotoxime and rosemary essential oil were determined. The ability to form biofilm as well as inhibition of biofilm formation of K. pneumoniae was performed. MICs 128, 0.25, 768, 64, 384 and 10 µg/ml were used. The effect of MIC and 1/2 MIC of antibiotics and rosemary essential oil

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Publication Date
Fri Aug 30 2024
Journal Name
Iraqi Journal Of Science
Investigation of Flagellum genes FleN and FlgE and Gene Expression of FleN Gene in Pseudomonas Aeruginosa Clinical Isolates
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The opportunistic multidrug resistance pathogen Pseudomonas aeruginosa has one or several flagella, and the numbers of these sophisticated machines are regulated by the flagellar regulator gene FleN. The flagellar hook gene FlgE is important for its synthesis, motility and tolerance to antibiotics. Bacteriahave resistance to antibiotics, especially to cephalosporin beta-lactam antibiotics. For the current study, 102 clinical specimens were collected and identified using routine laboratory tests and confirmed by Vitek-2 compact system.  A total of 33 isolates of P. aeruginosa were identified. The antibiotic susceptibility test was done by the Vitek 2 Compact system. Flagellar gene detected by conventional PCR revealed that the FleN

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Publication Date
Tue Aug 08 2023
Journal Name
Pharmaceutical Sciences Asia
Sub-inhibitory doses of Ofloxacin reduce adhesion and biofilm formation of Pseudomonas aeruginosa to biotic and abiotic surfaces
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Publication Date
Mon Apr 15 2019
Journal Name
Journal Of Global Pharma Technology
The dual role of Gentamicin in gene expression of fibronectin binding proteins in Staphylococcus aureus
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Objectives: The current work aimed to reveal the impact of gentamicin on the fibronectin binding proteins (fnbp) gene expression and its relation to biofilm and agr type in Staphylococcus aureus. Materials and Methods: A total of 25 S. aureus isolates were enrolled in this study previously isolated from different specimens. Identification confirmation and methicillin resistance were achieved by amplification of 16SrRNA and mecA. Multiplex polymerase chain reaction (PCR) based assay was employed to evaluate the agr typing. The gene expression of fnbA and fnbB genes was tested by real-time PCR technique. Minimum inhibitory concentration was estimated by micro broth dilution methodology. Microtiter plate method was performed to determine the a

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