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bsj-3583
Molecular Analysis of Rifampicin Resistance Conferring Mutations in Mycobacterium tuberculosis

Mycobacterium 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 rpoC genes. Twenty-three isolates (32.85%) showed resistance to rifampicin by either proportion method or Genexpert assay. Sequence analysis of the rpoB gene revealed fourteen different mutation patterns. Inside the rifampicin resistance determining region (RRDR), codons: S531L, D516V were highly mutated with frequencies of (21.73%, 17.39%) respectively. Outside the RRDR, there were nine different types of mutations, and M479L was the most prevalent one. Out of 23 RIF resistant isolates, seven isolates (30.43%) carried mutations in the rpoA gene, and twelve isolates (52.17%) harbored a mutation in rpoC. Most of the mutations were identified for the first time in this study. The current study demonstrated that mutations in rpoB, rpoA, and rpoC contributed to RIF resistance in Mycobacterium tuberculosis and this new finding may be relevant to realize how compensatory mutations in the rpoA and rpoC genes restore the fitness cost caused by rifampin resistance-conferring mutations in rpoB.

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Publication Date
Fri Jan 26 2024
Journal Name
Iraqi Journal Of Science
Induce mutations for Bavistin resistance in Trichoderma harzianum by UV-irradation

The present study was conducted to induce mutations by UV- light in Trichoderma harzianum for tolerance of Bavistin fungicides which are commonly used to control soil born phytopathogens. The mycelium growth of T. harzianum T1, T2 was inhibited completely by 30 μg (a.i.) /ml PDA of Bavistin. Two isolates of Trichoderma harzianum T1 and T2 conidia suspensions 104 conidia / ml were irradiated by UV lights at 254, 320, 365nm wavelength for 10, 20, 40, 80, 160 min using UV dark room cabinet and spectrophotometer and cultured on PDA media containing 50, 60, 75, 100, and 150 μg (a.i) / ml Bavistin. A thirty one unstable mutants tolerant to Bavistin were obtained after irradiation the T. harzianum T1, T2 conidia suspension 104 conidia / ml by

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Publication Date
Mon Jan 01 2024
Journal Name
Baghdad Science Journal
Molecular Identification of Methylorubrum extorquens using PCR-Amplified MxaF Gene Fragments as A Molecular Marker

  Methylotrophs bacteria are ubiquitous, and they have the ability to consume single carbon (C1) which makes them biological conversion machines. It is the first study to find facultative methylotrophic bacteria in contaminated soils in Iraq. Conventional PCR was employed to amplify MxaF that encodes methanol dehydrogenase enzyme. DNA templates were extracted from bacteria isolated from five contaminated sites in Basra. The gene specific PCR detected Methylorubrum extorquens as the most dominant species in these environments. The ability of M. extorquens to degrade aliphatic hydrocarbons compound was tested at the laboratory. Within 7 days, gas chromatographic (GC) studies of remaining utilize

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Publication Date
Sun Oct 02 2016
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Novel methods to design wild bacteriophages into highly lytic and therapeutic bacteriophages to extensively drug-resistant Mycobacterium tuberculosis

Background: The emergence of multiple drug resistant (MDR) Mycobacterium tuberculosis (M.TB) and extensive drug resistant (XDR) M.TB lay huge burden on TB endemic countries such as Iraq.
Objectives: Bacteriophage (phage) therapy can be used as alternative approach to tackle this problem. Patients and methods: Forty isolates of M.TB were cultured from TB-positive sputum specimens with three ATCC strains. Phage passaging and biokinetic based techniques were used to optimize wild anti-M.TB phages. Three chemical, non genetic- designing techniques, tween-80, mycobacterial lysis buffer, and xyelen, were used to change the specificity of wild anti-TB phages towards phage-resistant target M.TB.
Results: Five wild anti-M.TB phages were iso

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Publication Date
Mon Nov 14 2022
Journal Name
Biomedicine
Molecular characterization of HBB gene mutations in beta-thalassemia patients of Southern Iraq

Introduction and Aim: Beta-thalassemia is a serious inherited genetic disorder and an increasing health burden globally. Beta -thalassemia is caused by genetic globin abnormalities within the hemoglobin beta (HBB) gene. This study aimed to characterize the HBB gene mutations in beta -thalassemia among southern Iraqi patients. Materials and Methods: The study included 30 beta -thalassemia patients referred to the Thi-Qar Center for Genetic Diseases, Iraq and 15 control samples from a random group of apparently healthy individuals. Genomic DNA was isolated from blood sample collected from each individual. The DNA was amplified for specific regions of the HBB gene and the amplified products sequenced. The sequences generated were analysed for

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Publication Date
Tue Jun 20 2023
Journal Name
Baghdad Science Journal
Determination of Molecular Weight and Viscosity of Irradiated Polystyrene

 

The varied applications of polystyrene in various fields of life led to examining the cause of radiation influence on some rheological behavior of commercial Polystyrene (PS) solution in the chloroform (CHCl3) solvent. Polystyrene grains shape samples were irradiated using the radioactive element Cesium- 137 with (9 µci) activity for 10, 20, and 30 minutes. The viscosity of the polymer solution depends on the concentration and size (i.e. molecular weight) of the dissolved polymer. Experimental data showed that the radiation dose affected the value of viscosity (shear, relative, specific, and reduced). The viscosity value significantly reduced at 10 min radiation dose and when increasing the dose, the viscosity value increased

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Publication Date
Wed Mar 10 2021
Journal Name
Baghdad Science Journal
Molecular Typing of Two Suspected Cutaneous Leishmaniasis Isolates in Baghdad

Leishmaniasis is a group of parasitic diseases caused by Leishmania spp., an endemic infectious agent in developing countries, including Iraq. Diagnosis of cutaneous lesion by stained smears, serology or histopathology are inaccurate and unable to detect the species of Leishmania. Here, two molecular typing methods were examined to identify the promastigotes of suspected cutaneous leishmaniasis samples, on a species level. The first was species-specific B6-PCR and the second was ITS1-PCR followed by restriction fragment length polymorphism (RFLP) using restriction enzyme HaeIII. DNA was extracted from in vitro promastigote culture followed by amplification of kDNA by B6 or amplification and digestion of LITSR/L

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Publication Date
Tue Mar 30 2021
Journal Name
Baghdad Science Journal
Diversity Assessment by Molecular Barcoding and Seed Morphology in Ricinus communis L.: Molecular diversity of Ricinus seeds

Fourteen morphologically varied Ricinus communis L. seeds were collected from different localities in Egypt, El-Sudan and Saudi Arabia. Seed morphology and ITS barcoding analysis were performed to assess their diversity and phylogenetic relationship. Sequence’s alignment of nrITS region from different accessions display high levels of genetic similarities. Cluster analysis could not group different accessions according to their geographical distribution. Nevertheless, the genetic barcodes are interestingly matched with the morphological features of the Ricinus seeds. In conclusion, seed morphology proved to be a valuable tool in evaluating biodiversity and phylogenetic relationship in plant species with different loca

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Publication Date
Mon Jul 01 2013
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Molecular detection of Epstein Barr Virus in Women with Breast cancer

Background: Epstein Barr Virus (EBV) infection has been implicated in pathogenesis of several types of carcinomas such as nasopharyngeal carcinoma, gastric cancer and bladder cancer and has recently been associated with breast cancer.
Objective: To evaluate the relations between Epstein Barr virus-encoded small RNA (EBER) and breast cancer.
Methods: Twenty two cases of breast cancer were retrieved from the Al-Kadhimiya Teaching Hospital in Baghdad. Clinical data were analyzed from the medical records and formalin fixed, paraffin embedded tumor tissue were examined by Chromogeneic in situ hybridization (ISH) technique for the detection of EBER.
Results: The expression of EBER in tissues patients with breast cancer in the present

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Publication Date
Wed Apr 01 2015
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Molecular detection of ALS1 virulence gene of Candida albicans isolated from groups of Iraqi patients

Background: Candida albicans is the principal fungal infectious agent in human infection. Adhesion is thought to be an essential step for colonization and establishment of Candida infections.
Objectives: Identification and comparison of ALS1 virulence gene of adhesion family among different isolates of Candida albicans by PCR.
Patients and methods: One hundred eight samples were collected from different group of Iraqi patients. All samples were culture on Sabouraud′s agar, CHROMagar for identification while API Candida kit confirmatory test and extracted DNA was done for just Candida albicans isolates, detected the ALS1 gene, extracted RNA for synthesis of cDNA and detected of gene and compare between iso

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Publication Date
Wed Mar 25 2020
Journal Name
International Journal Of Drug Delivery Technology
Detection of Single Nucleotide Polymorphisms (SNPs) for Genes Cause Drug-Resistant in Iraqi Mycobacterium Tuberculosis isolates by new Pyrophosphate Technique.

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

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