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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
Wed Dec 01 2021
Journal Name
Iraqi Journal Of Veterinary Sciences
Isolation and antimicrobial resistance of Staphylococcus spp., enteric bacteria and Pseudomonas spp. associated with respiratory tract infections of sheep
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Sheep are considered as an important part of livestock in the worldwide, particularly in Iraq, as they provide meat, milk, leather, wool, and manure. The present study aim is isolation and identification of staphylococci, enteric bacteria and Pseudomonas spp. Totally, 115 samples were collected from sheep (100 samples were collected from the nasal cavity of local sheep suffering from respiratory infections, and 15 samples were collected from apparently healthy local sheep). All the samples were collected from seven flocks located in Abu Ghraib and Al-Radwaniyah, Baghdad governorate, Iraq. The samples were taken during the period from October 2020 to February 2021. Staphylococcus spp., Pseudomonas spp., and enteric bacteria were detected fi

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Publication Date
Mon Dec 31 2018
Journal Name
Iraqi Journal Of Market Research And Consumer Protection
STUDYING THE EFFECT OF Proteinase K AND α-Amylase ON INHIBITION OF Staphylococcus aureus AND Klebsiella pneumoniae BIOFILM ISOLATE FROM URINARY TRACT INFECTION.: STUDYING THE EFFECT OF Proteinase K AND α-Amylase ON INHIBITION OF Staphylococcus aureus AND Klebsiella pneumoniae BIOFILM ISOLATE FROM URINARY TRACT INFECTION.
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The current study aimed to investigate the viability of biofilm formation klebsilla pneumoniae and Staphylococcus aureus. 440 urine samples were collected from patients suffering from urinary tract infection (UTI) from those who were admitted and visitors to Al-Ramadi Teaching Hospital, Al-Yarmouk Teaching Hospital, Al-Ramadi Teaching Hospital for women and children and , Teaching Laboratories in the Medical City for both genders for a period extended from 5 July, 2017 to 10 October, 2017. Samples were diagnosed by culturing them on a selective media and by biochemical testes , also, diagnosis was ensured by using VITEK-2 compact system. Results showed that K.pneumoniae isolation ratio was 17.1%(68) and S.aureus ratio was 13.1%(52). Thei

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Publication Date
Mon Jan 15 2018
Journal Name
Iraqi Journal Of Laser
Effect of 410 nm Diode Laser Irradiation on the Growth of Burn Wounds-associated Bacteria, Pseudomonas Aeruginosa and Staphylococcus Aureus
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The effect of 410nm with 100 mW output power and one centimetre spot size (0.128 W/cm2 power density) Diode laser irradiation at different exposure times on the growth of Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus was evaluated. Seventy swap samples were collected from burn and infected wounds of 35 patients admitted to the burn-wound unit in Al-Yarmouk Teaching Hospital in Baghdad during the period from December 2014 to February 2015. These bacteria were isolated and identified depending on their growth on selective media, cultural characteristics, Gram stain morphology and biochemical tests and finally were confirmed by Vitek 2 compact system test .Susceptibility of bacterial isolates to 15antibiotics

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Publication Date
Tue Oct 20 2020
Journal Name
Ibn Al-haitham Journal For Pure And Applied Sciences
The impact of Glucose and Sodium Chloride on the Biofilm Formation of Pseudomonas aeruginosa & Staphylococcus aureus
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The aim of this research is to evaluate the effect of glucose and sodium chloride on biofilm formation by bacteria causing wound infection. For this purpose, 1% and 2% concentration of each of glucose and sodium chloride were used to test the biofilm formation potential of Staphylococcus aureus and Pseudomonas aeruginosa, which were the most common abundant bacteria that cause infection by biofilm. Each of the concentrations was kept in contact with the pathogenic bacteria for 24 hours. After the period of incubation, the concentration of 1% of glucose enhanced moderate biofilm formation capacity for (66% and 80%) on both bacteria respectively. The concentration of 2% glucose, on the other hand, led to a weak biofilm fo

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Publication Date
Sun Dec 09 2018
Journal Name
Baghdad Science Journal
Detection of Pseudomonas aeruginosa in Clinical Samples Using PCR Targeting ETA and gyrB Genes
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Pseudomonas aeruginosa has variety of virulence factors that contribute to its pathogenicity. Therefore, rapid detection with high accuracy and specificity is very important in the control of this pathogenic bacterium. To evaluate the accuracy and specificity of Polymerase Chain Reaction (PCR) assay, ETA and gyrB genes were targeted to detect pathogenic strains of P. aeruginosa. Seventy swab samples were taken from patients with infected wounds and burns in two hospitals in Erbil and Koya cities in Iraq. The isolates were traditionally identified using phenotypic methods, and DNA was extracted from the positive samples, to apply PCR using the species specific primers targeting ETA, the gene encoding for exotoxin A, and gyrB gene. The res

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Publication Date
Mon Apr 10 2017
Journal Name
Ibn Al-haitham Journal For Pure And Applied Sciences
Role of Extracted Genomic DNA on Biofilm Formation by Pseudomonas aeruginosa and Klebsiella pneumoniae in vitro
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   Bacteria form complex and highly elaborate surface adherent communities known as biofilms.Biofilm have been shown to be associated with several human diseases ,and to colonize a wide variety of medical devices . The current study focuses on contribution of extracted genomic   DNA in  biofilm formation by   P. aeruginosa and  K. pneumoniae isolates   .The percentages of Pseudomonas aeruginosa recovery from drinking water  in this study were  10%(20 positive P. aeruginosa  samples ) and K. pneumonia.,  7%(14 positive K. pneumonia samples).The results showed that all P.aeruginosa and K. pneumoniae isolates (100%) were slime producer but in different degrees by forming  of black

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Publication Date
Tue Jul 12 2016
Journal Name
World Journal Of Experimental Biosciences
Effect of honey and sodium bicarbonate on formation inhibition and removal of Staphylococcus aureus and Klebsiella pneumoniae biofilm in vitro
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Publication Date
Wed Feb 01 2023
Journal Name
Tropical Journal Of Natural Product Research
Expression of algD Gene in Single- and Dual-Species Biofilms of Pseudomonas aeruginosa and Staphylococcus aureus Under Starvation Stress
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Dual-species biofilms of Pseudomonas aeruginosa and Staphylococcus aureus generate difficult-to-treat illnesses. Nutrition stress in biofilms affects physiology, microbial metabolism, and species interactions, impacting bacteria growth and survival. Furthermore, the function of alginate, which is encoded by the algD gene, in the production of biofilms has been established. The present study aimed at investigating the impact of starvation on algD gene expression in single-species biofilm of P. aeruginosa and dual-species biofilms of P. aeruginosa and S. aureus from hospital sewage. A total of six P. aeruginosa and six S. aureus isolates were obtained from the microbiology laboratory at the Department of Biology, College of Science, Universit

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Publication Date
Sat Jan 01 2011
Journal Name
North American Journal Of Medical Sciences
Urinary tract infections caused by staphylococcus aureus DNA in comparison to the candida albicans DNA
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Background: Bacterial DNA released upon bacterial autolysis or killed by antibiotics, hence, many inflammatogenic reactions will be established leading to serious tissue damage. Aim: the present work aimed to elucidate the histopathological changes caused by prokaryotic (bacterial) DNA and eukaryotic (candidal) DNA. Materials and methods: twenty one Staphylococcus aureus and 36 Candida albicans isolates were isolated from UTI patients. Viable cells and DNA of the highest antibiotic sensitive isolates were injected, intraurethraly, in mice. Results were evaluated via histopathological examination. Results: Mildest reactions were obtained from mice challenged with viable C. albicans compared with those challenged with viable S. aureus. Dos

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Publication Date
Sat Sep 01 2018
Journal Name
Iraqi Journal Of Science
Estimate Antimicrobial activity and Anti-biofilm formation of bark Cinnamomum zeylanicum on Klebsiella pneumoniae isolated from Urinary Tract Infections
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