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.
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
... Show MoreThis study investigates in vitro biofilm production. Presence of ica A and D genes in methicillin-resistant Staphylococcus aureus was evaluated for biofilm production by the microtiter plate method. Between December 2020 and October 2021, out of 215 clinical specimens were collected from patients with pulmonary fibrosis, pneumonia, bacteremia, chronic burns, deep wounds, urinary tract infection and catheterized patients. Out of which 45 MRSA isolates were identified by the susceptibility test utilizing cefoxitin and the occurrence of mecA gene for resistance for this antibiotic verified by polymerase chain reaction technique. A sensitivity test was conducted for five other antibiotics
... Show More57 isolates of Mycobacterium tuberculosis and Mycobacterium bovis were identified; they were isolated from different clinical sources which included sputum, bronchial wash, abscess, pleural fluid, gastric fluid, eye fluid, and CSF, also urine and ear swab. This investigation was carried out on 198 patient attended National Reference Laboratory for T.B during September 2009. Also the study declared that the ratio of separation of this bacterium from male was (67.6%) and it’s higher than the ratio of separation this bacterium from females which was (32.3%). The susceptibility of Mycobacterium tuberculosis to melatonin was evaluated. Many concentrati
... Show MoreMycobacterium tuberculosis resistance to rifampicin is mainly mediated through mutations in the rpoB gene. The effects of rpoB mutations are relieved by secondary mutations in rpoA or rpoC genes. This study aims to identify mutations in rpoB, rpoA, and rpoC genes of Mycobacterium tuberculosis isolates and clarify their contribution to rifampicin resistance. Seventy isolates were identified by acid-fast bacilli smear, Genexpert assay, and growth on Lowenstein Jensen medium. Drug susceptibility, testing was performed by the proportional method. DNA extraction, PCR, and sequencing were accomplished for the entire rpoA, rpoB, and
... Show MoreIn this study, 25 clinical isolates of Proteus spp. were collected from urine, wounds and burns specimens from different hospitals in Baghdad city, all isolates were identified by using different bacteriological media, biochemical assays and Vitek-2 system. It was found that 15 (60%) isolates were identifies as Proteus mirabilis and 10 (40 %) isolates were Proteus vulgaris. The susceptibility of P. mirabilis and P. vulgaris isolates towards cefotaxime was (66.6 %) and (44.4 %) respectively; while the susceptibility of P. mirabilis and P. vulgaris isolates towards ceftazidime was (20%). Extended spectrum β-lactamses producing Proteus was (30.7 %). DNA of 10 isolates of P. mirabilis and 4 isolates of P. vulgaris were extracted and detecti
... Show MoreVariation in DNA, and genes to a lesser or greater extent, can play an important role in most diseases; that is because this variation in will reflect and affect the function of DNA, and genes (combined genes and DNA or separately). This can be affected by environment, life style, as well as the inheriting from parents and previous generations. All these factors can contribute in human diseases. There are different alterations in genes, like imbalance and inequality in chromosomes, disorder in gene (deficiency in gene, which could be complex or single disorder), and cancer. In the last decades, scientists were focus on medicine and genetics; they pay an extensive attention to reach better understanding about diseases and their cause
... Show MoreType 1 diabetes (T1D) is an autoimmune disease with chronic nature resulting from a combination of both factors genetic and environmental. The genetic contributors of T1D among Iraqis are unexplored enough. The study aimed to shed a light on the contribution between genetic variation of interleukin2 (IL2) gene to T1D as a risk influencer in a sample of Iraqi patients. The association between IL2−330 polymorphism (rs2069762) was investigated in 322 Iraqis (78 T1D patients and 244 volunteers as controls). Genotyping for the haplotypes using polymerase chain reaction test – specific sequence primer (PCR-SSP) for (GG, GT, and TT) genotypes corresponding to (G and T) alleles were performed. A significant association revealed a decreased freq
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