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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
Tue Jan 01 2013
Journal Name
The Iraqi Journal Of Veterinary Medicine
Detection of methicillin or multidrug resistant Staphylococcus aureus (MRSA) in locally produced raw milk and soft cheese in Baghdad markets.
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In order to investigate the presence of methicillin or multidrug resistant Staphylococcus aureus in food-chain especially Cows raw milk and white raw soft cheese and its whey, a total of 30 samples were collected randomly from different markets in Baghdad Province during December 2012 till February 2013, in which samples were analyzed by a standard isolation protocols of food microbiology with some modification processing by new, modern and rapid technology tools such as chromogenic medium Baird-Parker agar, Electronic RapIDTM Staph Plus Code Compendium Panel System (ERIC®) Dryspot Staphytect Plus and Penicillin Binding Protein (PBP2') Plus assays; as well as, studying the susceptibility of isolates to different selected antibiotics. The r

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Publication Date
Fri Aug 02 2024
Journal Name
Rafidian Journal
Quorum Sensing in Staphylococcus aureus
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Staphylococci are common commensals in human beings, yet certain species are pathogenic. Staphylococcus aureus, particularly, is a very virulent human pathogen. The capacity of staphylococci to sense the density of bacterial cell, i.e., quorum, and thereafter respond via genetic modifications is attributable to one primary mechanism known as accessory gene regulator (Agr). Agr's extracellular signal is a peptide that is posttranslationally modified with a thiolactone molecule. Agr is in charge of the upregulation of numerous exotoxins and hydrolyzing enzymes, as well as the downregulation of many colonization determinants, under circumstances of high cell density. This modulation is critical for the scheduling synthesis of virul

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Publication Date
Sun Jun 01 2014
Journal Name
Baghdad Science Journal
Identification Pseudomonas aeruginosa by 16s rRNA gene for Differentiation from Other Pseudomonas Species that isolated from Patients and environment
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Pseudomonas aeruginosa is a common and major opportunistic human pathogen, its causes many and dangersinfectious diseases due to death in some timesex: cystic fibrosis , wounds inflammation , burns inflammation , urinary tract infection , other many infections otitis external , Endocarditis , nosocomial infection and also causes other blood infections (Bacteremia). thereforebecomes founding fast and exact identification of P. aeruginosafrom samples culture very important.However, identification of this species may be problematic due to the marked phenotypic variabilitydemonstrated by samples isolates and the presence of other closely related species. To facilitate species identification, we used 16S ribosomal DNA(rRNA) sequence data

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Publication Date
Mon Oct 01 2018
Journal Name
International Journal Of Biosciences
Detection of Escherichia coli O157 H7 isolated from infected dogs with urinary tract infections using real time - polymerase chain reaction
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Publication Date
Wed Jan 01 2025
Journal Name
Medical Journal Of Babylon
Optimum Conditions for Pyomelanin Production and Characterization from Local Isolates of Pseudomonas aeruginosa
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Nowadays, there is increased interest in the biosynthesis of microbial melanin related to their numerous biological functions and applications in many fields, especially in medical fields, including immune-modulating, antimicrobial antibiotic, antiviral antivenin, anticancer, antitumor activity, and anti-biofilm activity. Pyomelanin is a hydrophobic macromolecule that is typically dark brown or black in color, formed by the oxidative polymerization of phenolic or indolic compounds. Pyomelanin is reported to be safe for consumption, thus providing a crucial strategy for biocontrol of biofilm. Furthermore, natural pyomelanin is known as a potent antioxidant, photoprotective, and free radical scavenging. Objective: This study focuses on the

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Publication Date
Fri Oct 02 2020
Journal Name
International Journal Of Pharmaceutical Research
Detection of Toxoplasma gondii in blood and milk of infected goats and Pregnant women by Rapid test cassette and conventional -PCR methods in AL-Qadisiyah province, Iraq
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Aim: The study aimed to investigate the presence of the specific B1 gene T gondii in blood and milk samples from natural infected cattle and pregnant women (16-30 weeks) whose examination performed by the officers at the women's and children's Educational hospital in Al-Diwaniyah, Iraq. Materials and methods: A total of 150 serum samplings were collected analysed and scanned for Anti-T gondi antibodies (75 naturally-infected goats and 75 pregnant women with Toxoplasma). Polymerase chain reaction (PCR) was used to detect of B1(399pb) gene in 26 goat's blood samples and 7 samples from pregnant women. Results: A quick-test anti-cassette gondii results showed 26 positive samples of goats in a percentage of 34,666 percent, while a higher percent

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Publication Date
Tue Jan 01 2019
Journal Name
Indian Journal Of Public Health Research & Development
Prevalence of Urinary Tract Infections in Adult and Child Patients
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Publication Date
Tue Feb 28 2023
Journal Name
Iraqi Journal Of Science
Gentamicin Upregulates the Gene Expression of hla and nuc in Staphylococcus aureus
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The current study aimed to detect the effect of gentamicin stress on the expression of hla (encodes hemolysin) and nuc (encodes nuclease) genes of Staphylococcus aureus. Fifty-eight isolates identified as S. aureus were isolated locally from different clinical specimens. Disk diffusion method was used to detect the resistance to S. aureus. The minimum inhibitory concentration (MIC) of gentamicin was estimated by broth microdilution method. hla and nuc genes were determined by polymerase chain reaction technique. The biofilm was evaluated using the microtiter plate method in the presence and absence of gentamicin at sub-MIC. The results showed that 18 (31%) and 40 (69%) S. aureus isolates were sensitive and resistant to gentamicin, respectiv

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Publication Date
Sun Sep 01 2019
Journal Name
Baghdad Science Journal
Detection a New Antiseptic Resistant Variant of qac Gene in Some Multi Drug Resistant Staphylococcus aureus Isolated from Different Clinical Sources
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The increasing use of antiseptic compounds creates selective pressure cause emergence of antiseptic resistance among Staphylococcus aureus .Resistance mechanism of antiseptic is driven mainly by multi drug resistant (MDR) efflux protein.Sixty five isolates of S.aureuswere collected from different clinical sources and subjected to 11 antibiotics most of them are recognized by efflux systems as extruded substrates. Range of efflux activity was estimated using cartwheel method. Simultaneous discrimination of antiseptic coding genes (qacA/B, smr and norA)as well as nuc and mecA genes among multidrug resistantS.aureus(MRSA) isolates was preformed using multiplex PCR assay

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Publication Date
Sun May 10 2020
Journal Name
Baghdad Science Journal
Prevalence of Quinolones Resistance Proteins Encoding Genes (qnr genes) and Co-Resistance with β-lactams among Klebsiella pneumoniae Isolates from Iraqi Patients
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This study investigated the prevalence of quinolones resistance proteins encoding genes (qnr genes) and co-resistance for fluoroquinolones and β-lactams among clinical isolates of Klebsiella pneumoniae.  Out of 150 clinical samples, 50 isolates of K. pneumoniae were identified according to morphological and biochemical properties. These isolates were collected from different clinical samples, including 15 (30%) urine, 12 (24%) blood, 9 (18%) sputum, 9 (18%) wound, and 5 (10%) burn. The minimum inhibitory concentrations (MICs) assay revealed that 15 (30%) of isolates were resistant to ciprofloxacin (≥4µg/ml), 11 (22%) of isolates were resistant to levofloxacin (≥8 µg/ml), 21 (42%) of isolates were re

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