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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 Feb 28 2023
Journal Name
Iraqi Journal Of Science
The Ability of Staphylococcus aureus to Establish Biofilm on Acrylic, Plastic, and Metallic Denture Materials
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       Staphylococcus aureus is a common pathogenic agent due to its ability to cause various types of infections, ranging from mild skin infections to sever systemic diseases. One of the most virulence factors of this bacterium is its ability to from biofilms on solid surfaces by anchoring the planktonic cells and by producing a protective layer of extra polymeric substances. Biofilm formation is controlled through many genes. The most important ones are icaA and icaD. Dentures are prosthetic devices that are made of different materials to replace lost teeth. The aim of this study is to examine the ability of different types of denture materials to support the biofilm formation of S. aureus at phenotypic level by detecting ba

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Publication Date
Wed Sep 21 2022
Journal Name
Biochemical And Cellular Archives
ANTIBIOTIC SUSCEPTIBILITY AND BIOFILM FORMATION OF PSEUDOMONAS AERUGINOSA ISOLATED FROM CLINICAL AND ENVIRONMENTAL HOSPITAL SAMPLES
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Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen and a model bacterium for studying virulence and bacterial social traits. While it can be isolated in low numbers from a wide variety of environments including soil and water, it can readily be found in almost any human/animal-impacted environment. It is a major cause of illness and death in humans with immunosuppressive and chronic conditions, and infections in these patients are difficult to treat due to a number of antibiotic resistance mechanisms and the organism’s propensity to form multicellular biofilms. One hundred twenty clinical samples and forty hospital environmental samples (various sources) were collected from hospitals in Baghdad city during the period from Oc

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Publication Date
Tue Dec 01 2020
Journal Name
Meta Gene
Association between the rs2234671 polymorphism and the risk of recurrent urinary tract infections in Iraqi women
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A case-control study was designed to find out the association between rs2234671 polymorphism of cxcr1 and rUTI in a sample of Iraqi women by polymerase chain reaction- sequence-specific primer (PCR-SSP) method. The current findings revealed that the genotype GC (OR= 7.86, 95% CI = 2.82-21.87, P= 7.7 × 10-5) and the C allele (OR= 3.93, 95% CI = 1.97 - 7.83, P = 9.8×10-5) are significantly associated with rUTI. However, the genotype GG played as a protective factor (OR= 0.12, 95% CI = 10.05 - 0.34, P = 4.0 ×10-5). Depending on these findings, the genotype GC is significantly associated with rUTI.

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Publication Date
Sun Jun 01 2014
Journal Name
Baghdad Science Journal
Study of Some Physical and Chemical Properties of Staphylolysin Enzyme purified from Pseudomonas aeruginosa
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Some of the characters of the Staphylolysin A and D enzymes purified from Pseudomonas aeruginosa P16 and P5 respectively were studied, the molecular weights of Staphylolysin A and D were 20.417 kilo dalton and 23.988 kilo Dalton respectively by SDS- polyacryl amide gel electrophoresis. The optimum pH for staphylolysin A activity was found to be 8 which gives higher activity reaches 150 unit/ml, and for enzyme stability was 7.5-8.5 in which the enzyme nearly retained its full activity, while it was 9.5 for staphylolysin D that gives higher activity of 16 unit/ml,and 8.5-9.5 for enzyme stability in which the enzyme nearly retained its full activity, Maximum activity of two enzymes was obtained at 40C in which the specific activity for st

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Publication Date
Thu Jun 30 2016
Journal Name
Al-kindy College Medical Journal
Antibacterial activity of Trigonella Foenum- groecum essential oil against skin infection with Staphylococcus aureus: In vitro and in vivo studies
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Background: Multi- drug resistant (MDR) Staphylococcus aureus infections have become a major public health concern in both hospital and community settings.Objective: to investigate the antibacterial activity of T. Foenum- groecum essential oil against skin infection with S. aureus and to study probable synergistic activity in combination with Clindamycin.Type of the study: Cross-sectional study.

Methods: Antibacterial activity of T. Foenum- groecum essential oil extract (1.2gm/100 µl) was investigated in multi- drug resistance (MDR) Staphylococcus aureus specimen isolated from patients with skin infection in Baghdad. T. Foenum- groecum use externally for cellulites and skin inflammation due to the presence of diosgenin .fast liq

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Publication Date
Mon Nov 14 2022
Journal Name
Biomedicine
Antibacterial activity of klebocin against methicillin resistance Staphylococcus aureus
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  Introduction and Aim: Bacteriocins are antimicrobial peptides that have bactericidal and/or bacteriostatic activity against other bacteria. The aim of this study was to assess the antibacterial efficiency of Klebocin a K. pneumoniae bacteriocin, against biofilm formation by clinical isolates of methicillin resistant Staphylococcus aureus MRSA.   Materials and Methods: S. aureus isolated from clinical samples was identified according to vitek 2 system Antibiotic susceptibility test was performed according to disc diffusion method. Vitek 2 compact system was also used to detect MRSA strains. Agar well diffusion method was used to evaluate the antibacterial activity of klebocin from K. pneumoniae towards 11 strains of S. aureus by

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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
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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Publication Date
Tue Dec 20 2022
Journal Name
Iraqi Journal Of Biotechnology
Synergistic Effect of Conocarpus erectus Extract and some Antibiotics against Multi-Drug Resistant Pseudomonas aeruginosa
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Over the past few decades, the health benefits are under threat as many commonly used antibiotics have become less and less effective against certain illnesses not only because many of them produce toxic reactions but also due to the emergence of drug-resistant bacteria. The clinical use of a combination of antibiotic therapy for Pseudomonas aeruginosa infections is probably more effective than monotherapy. The present study aims to estimate the antibacterial and antibiofilm activity of Conocarpus erectus leaves extracts against multi-drug resistant P. aeruginosa isolated from different hospitals in Baghdad city. One hundred fifty different clinical specimens were collected from patients from September 2021 to January 2022. All samples were

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Publication Date
Wed Feb 06 2019
Journal Name
National Academy Science Letters
The Detection Limit of PCR Amplification for Cryptosporidium spp. Oocysts in Fecal Samples
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