In this search, a new bioluminescent technique was proved for pyrophosphate which was employed to single- nucleotide polymorphism (SNP) diagnosis using one-base extension reaction. Four Mycobacterium tuberculosis genes were chosen (Rpob, InhA, KatG, GyrA) 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 procedure of this assay was as follows. A specific primer within each aliquot owning a short 3-OH end of the base of the target gene was hybridized to the single-stranded DNA template. Then, (exo-) Klenow DNA polymerase and one of either ?-thio-dATP, dTTP, dGTP, or dCTP were supplemented and incubated for 1 min. Pyrophosphate freed by DNA polymerase is altered to ATP by pyruvate phosphate dikinase (PPDK), and the amount of ATP is measured using the firefly luciferase reaction. This technique, which does not demand expensive equipment, can be applied to rapidly monitor 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
A new design of manifold flow injection (FI) coupling with a merging zone technique was studied for sulfamethoxazole determination spectrophotometrically. The semiautomated FI method has many advantages such as being fast, simple, highly accurate, economical with high throughput . The suggested method based on the production of the orange- colored compound of SMZ with (NQS)1,2-Naphthoquinone-4-Sulphonic acid Sodium salt in alkaline media NaOH at λmax 496nm.The linearity range of sulfamethoxazole was 3-100 μg. mL-1, with (LOD) was 0.593 μg. mL-1 and the RSD% is about 1.25 and the recovery is 100.73%. All various physical and chemical parameters that have an effect on the stability and development of
... Show MoreTuberculosis (TB) still remains an important medical problem due to high levels of morbidity and mortality worldwide. A series of innate immune mechanisms that create a cytokine network control the pathogenesis of tuberculosis and this response has the capacity to modify the host genomic DNA structure through epigenetic mechanisms such as DNA methylation which could constantly alter the local gene expression pattern that can modulate the metabolism of the tissues and the immune-response. Interferon-gamma (IFN-γ) is an important pro-inflammatory cytokine regulator of the innate immune response to TB. This study aims to determine DNA methylation patterns of INF-γ gene promoter and measure serum IFN- γ level in newly diagnosed TB patient
... Show MoreNon tuberculous mycobacteria (NTM) present in the environment, mainly in water and not transmitted from human to human. The lung is the most common target of NTM infections in human. The symptoms of NTM infection are: fever, weight loss and abdominal pain.
114 sputum samples were examined microscopically using Ziehl-Nelsen stain, all were positive. They cultured on Lowenstein-Jensen media.
The results of this study indicate presence of 3 (2.6%) of rapidly growing mycobacteria, phenotypically they resemble T.B. bacilli. They were diagnosed by biochemical tests, they were positive for catalase and pyruvate, negative for niacin, nitrate reduction test, sensitive for Thiophene-2-carboxyl acid hydrozide test& resistant for Para nitr
One hundred thirty - five clinical specimens of urine, blood, teeth root canal and burns were obtained from patients in hospitals of Baghdad. The specimens were cultured on Pfizer Selective Enterococcus agar to purify Enterococci isolates. 20 E. faecalis isolates were identified biochemically by growing in 10Cº, 45Cº, 6.5% NaCl, at pH 9.6 and confirmed by VITEK. Determination of Vancomycin-Resistant E. faecalis isolates were done by the minimum inhibitory concentrations [MICs] using agar dilution method. Seventeen E. faecalis isolates were determined as Vancomycin-Resistant and Intermediate Resistant.
Susceptibility of thirty seven clinical isolates of Staphylococcus aureus to various antibiotics was tested. 100 % of tested isolates were resistant to ampicillin, while the lowest resistance recorded to amikacin 8.10 %. Four of S. aureus isolates showed resistant to vancomycin. Minimum inhibitory concentration (MIC) of isolates 33 and 56 for vancomycin was ≥ 32 μg/ml.
One hundred thirty - five clinical specimens of urine, blood, teeth root canal and burns were obtained from patients in hospitals of Baghdad. The specimens were cultured on Pfizer Selective Enterococcus agar to purify Enterococci isolates. 20 E. faecalis isolates were identified biochemically by growing in 10Cº, 45Cº, 6.5% NaCl, at pH 9.6 and confirmed by VITEK. Determination of Vancomycin-Resistant E. faecalis isolates were done by the minimum inhibitory concentrations [MICs] using agar dilution method. Seventeen E. faecalis isolates were determined as Vancomycin-Resistant and Intermediate Resistant.
One hundred and six S. aureus were isolated from 250 Nasal swabs of
Healthcare workers and patients at Al- Kadhamia teaching Hospital and Al-
Numan hospital, Baghdad, Iraq. The study was undertaken over a period of
ten months between August 2011 and June 2012. S. aureus isolates were
diagnosed based on phenotypic traits and biochemical tests. Antibiotics
sensitivity to 11 antibiotics, revealed that S.aureus is totally resistant to
Pencillin G (100%), highly resistant to Cefoxitin (alternative to Methicillin)
(94.3%) While there are varied resistance percentage for the rest of
antibiotics: Erythromycin (37.7%), Tetracycline (34.9%), Gentamicin
(29.3%), Trimethoprim/sulfamethoxazole (50%), Ciprofloxacin (29.2%),<
A method is developed for the determination of iron (III) in pharmaceutical preparations by coupling cloud point extraction (CPE) and UV-Vis spectrophotometry. The method is based on the reaction of Fe(III) with excess drug ciprofloxacin (CIPRO) in dilute H2SO4, forming a hydrophobic Fe(III)- CIPRO complex which can be extracted into a non-ionic surfactant Triton X-114, and iron ions are determined spectrophotometrically at absorption maximum of 437 nm. Several variables which impact on the extraction and determination of Fe (III) are optimized in order to maximize the extraction efficiency and improve the sensitivity of the method. The interferences study is also considered to check the accuracy of the procedure. The results hav
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