In this search, a new pyrophosphate technique was proved. The technique was employed to single- nucleotide polymorphisms (SNPs), which diagnosis using a one-base extension reaction. Three Mycobacterium tuberculosis genes were chosen (Rpob, InhA, KatG) genes. Fifty-four specimens were used in this study fifty-three proved as drug-resistant specimens by The Iraqi Institute of Chest and Respiratory Diseases in Baghdad.; also one specimen was used as a negative control. The steps of this technique were by used a specific primer within each aliquot that has a short 3-OH end of the base of the target gene that was hybridized to the single-stranded DNA template. Then, the Taq polymerase enzyme and one of either α-thio-dATP, dTTP, dGTP, or dCTP were supplemented and incubated for 1 min. ATP is synthesis by convert Pyrophosphate freed by DNA polymerase using pyruvate phosphate dikinase (PPDK), and the amount of ATP estimates by the firefly luciferase reaction. This technique, which does not demand expensive equipment, can be applied to rapidly monitor a one-point mutation in the gene that causes drug-resistant in mycobacterium tuberculosis. The results showed a high variation in values of ATP formation through matching and mismatch bases added. So, this assay (which required only five minutes), enable to find the gene SNP causes resistant for the specific drug.
One hundred forty three of Klebsiellapneumoniae isolates had been collected from some hospitals in Baghdad city. The isolates were taken from different clinical specimens.Antimicrobial susceptibility test was carried out towards fifteen antimicrobial agents by using Vitek2 system with Antimicrobial susceptibility test cards. The results of antibiogram showed that the local isolates were possess highly resistance towards most antimicrobial agents under study. The high resistance wastoAmpicillin while the low resistance was to Imipenem.Two methods were used for detection of Extended Spectrum Beta Lactamases (ESBLs) production; first methods by using of Vitek2 system,thesecondmethods by using of polymerase chain reaction (PCR) technique to dis
... Show MoreThis paper demonstrates a new technique based on a combined form of the new transform method with homotopy perturbation method to find the suitable accurate solution of autonomous Equations with initial condition. This technique is called the transform homotopy perturbation method (THPM). It can be used to solve the problems without resorting to the frequency domain.The implementation of the suggested method demonstrates the usefulness in finding exact solution for linear and nonlinear problems. The practical results show the efficiency and reliability of technique and easier implemented than HPM in finding exact solutions.Finally, all algorithms in this paper implemented in MATLAB version 7.12.
MRSA is one of the major pathogens in hospitals and the community, which have the ability to produce biofilm as a virulence factor, the impact of chalcone on biofilm formation, the synergism effect of chalcone and antibiotic in both in vitro and in vivo experiments, the gene expression of virulence genes (srtA, fnbA, fnbB) before and after treatment of it on MRSA biofilm cells in vitro, all these were the prime aims of this study. Chalcone at MBIC (20 μg/ml), significantly reduced the biofilm formation to 21.45% and at sub MBIC (15 μg/ml) to 36.58 %. While, Chalcone at MIC(5 μg/ml) reduced MRSA planktonic cells to 49.61%. Susceptibility of MRSA isolates against eight antibiotics showed that all isolates were sensitive to vancomycin and n
... Show MoreSGI2 wild type isolate of Sinorhizobium meliloti was isolated from Medicago sativa (alfalfa) plant which was obtained from Al-Tarmiaa region / Baghdad. Nine auxotrophic mutants were obtained from the SGI2 wild type isolate by mutagenesis with Nitrous acid (HNO2). The SGI2 wild type and the all auxotrophic mutant isolates had two Megaplasmids; pSymA and pSymB. No genetic variations in plasmid number and size were detected when gel electrophoresis was done for plasmid profile detection. Genetic variations by using RAPD-PCR technique were obtained between wild type and auxotrophic mutant isolates. One band was detected in SGI6 gel profile with 1.5 Kb size when OPY-04 primer was used. Using OPB7 primer by using RAPD-PCR technique showed larg
... Show MoreThe current work aims to evaluate the association between genetic mutations in thymidylate synthetase (
For the period from February 2014 till May 2014, one hundred and nine lactose fermenter clinical isolates from different samples (urine, stool, wound swab, blood, and sputum) were collected from Alyarmok, Alkadimiya, and Baghdad teaching hospitals at Baghdad governorate. Identification of all Klebsiella pneumoniae isolates were carried out depending on macroscopic, microscopic characterizations, conventional biochemical tests, and Api 20E system. Fifty-three (48.62%) isolates represented K. pneumoniae; however, 51.73% represented other bacteria. Susceptibility test was achieved to all fifty-three K. pneumoniae isolates using five antibiotic disks (Ceftazidime, Ceftriaxone, Cefotaxime, Imipenem, and Meropenem). Most of tested isolates (90
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