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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
Thu Nov 14 2024
Journal Name
Journal Of Emergency Medicine, Trauma And Acute Care
Isolation, optimization, and redesigning of phages of methicillin-resistant Staphylococcus aureus from clinical hospital isolates in Baghdad
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Background: A global health concern is methicillin-resistant Staphylococcus aureus (MRSA). The use of bacteriophages is one of the many novel control strategies against MRSA that are frequently sought. However, it is quite challenging to isolate enough lytic anti-MRSA phages. In order to extract, optimize, and remodel anti-MRSA phages, this study sought novel approaches.

Methods: Two ATCC MRSA strains and nine clinical MRSA isolates were used to isolate wild anti-MRSA phages from hospital settings, dirt, and sewage. The wild phages were optimized using plaque-based biokinetic techniques. Usi

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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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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
Wed Jan 01 2020
Journal Name
Systematic Reviews In Pharmacy
Impacts of the alcoholic extract and essential oil of Thymus vulgaris L. Against the causative agent of acne formation (Staphylococcus aureus)
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Medicinal plant life have performed an integral role in the development of human lifestyle being used as resources for the manufacturing of contemporary medicines, antibiotics. Microorganisms are responsible for many issues in industry and remedy because of biofilm formation. This study aimed to take a look at the results of Thymus vulgaris alcoholic extract on Staphylococcus aureus the causative marketers of acne in human particularly the women. The extract was acquired mixing 25 g of Thyme powder with 350 ml of 80% ethanol for 6 hours at 40°. Antimicrobial activities of the plant extract and integral oil towards the S. aureus were decided using the agar diffusion technique. The consequences proven that special concentrations of alcoholic

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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
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
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
Sun Jul 29 2018
Journal Name
Iraqi Journal Of Science
Incidence of Ciprofloxacin-Resistant of Methicillin Resistant Staphylococcus aureus isolated from Iraqi patients
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The resistance of Staphylococcus aureus to ciprofloxacin has complicated the problem of treating staphylococcal associated infections in which MRSA is the causative agent since ciprofloxacin was the drug of choice to treat such infections. Our study investigated the incidence of Ciprofloxacin resistant S. aureus isolates that were also methicillin resistant among Iraqi patients. The obtained bacterial isolates were tested for Ciprofloxacin resistance using agar dilution method and the sequence of gyrA and parC. The results revealed that about 8% of the isolated MRSA strains were Ciprofloxacin resistant and the resistance was due to mutation in gyrA rather than parC.

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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