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
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
... Show MoreBackground: 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
57 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 MoreBackground: Tuberculosis (TB) is a major public health issue and a main cause of global morbidity and mortality. TB is the world's ninth leading cause of death despite the numerous treatment strategies for managing the disease.
Objective: To assess the traditional method (direct smear examination and culture) against real-time PCR as pulmonary and extrapulmonary tuberculosis laboratory diagnostic techniques.
Cases and methods: Samples were collected from (612) TB cases, (409) of whom were pulmonary tuberculosis (PTB) and (203) were extrapolmonary tuberculosis (EPTB). The cases were seeking care at the Specialized Chest and Respiratory Disease Center/ National Reference Laboratory for Tuberculosis (NRL) in Baghdad, during the period
Tuberculosis status as the second leading causes of significant morbidity and mortality from an infectious disease worldwide, after human immunodeficiency virus (HIV). Sample collection was conducted at the Institute of Chest and Respiratory Diseases/Baghdad Medical City in Baghdad. The collection interval was from August to October 2014, 629 suspected TB patients were examined during this period. The results revealed among total 629 specimens, 56 (8.9%) of the specimens were positive by direct examination and 573 (91.1%) negative specimens by smear microscopy. Fifty six DNA samples were extracted from positive ZN smears of sputum specimens and 40 samples from healthy persons (as control) were subjected to molecular diagnosis by real tim
... Show MoreFraud Includes acts involving the exercise of deception by multiple parties inside and outside companies in order to obtain economic benefits against the harm to those companies, as they are to commit fraud upon the availability of three factors which represented by the existence of opportunities, motivation, and rationalization. Fraud detecting require necessity of indications the possibility of its existence. Here, Benford’s law can play an important role in direct the light towards the possibility of the existence of financial fraud in the accounting records of the company, which provides the required effort and time for detect fraud and prevent it.
G-system composed of three isolates G3 ( Bacillus),G12 ( Arthrobacter )and G27 ( Brevibacterium) was used to detect the mutagenicity of the anticancer drug, cyclophosphamide (CP) under conditions similar to that used for standard mutagen, Nitrosoguanidine (NTG). The CP effected the survival fraction of isolates after treatment for 15 mins using gradual increasing concentrations, but at less extent comparing to NTG. The mutagenic effect of CP was at higher level than that of NTG when using streptomycin as a genetic marker, but the situation was reversed when using rifampicin resistant as a report marker. The latter effect appeared upon recording the mutagen efficiency (ie., number of induced mutants/microgram of mutagen). Measuring the R
... Show More