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.
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
... Show MoreGenetic and environmental factors are believed to have a key role in the development and pathogenesis of autoimmune thyroid diseases (AITD). This study aimed to investigate the association between two CTLA-4 gene single nucleotide polymorphisms (SNPs) CT60/rs3087243 and CT61/rs11571319 with autoimmune thyroiditis in a sample of Iraqi patients. Seventy-five patients (67 females, 8 males) and eighty-eight subjects (79 females and 9 males) matched in age, gender, and ethnicity as a control group. Thyroid autoantibodies were present in females more than in males with a total positivity of anti-TPO of 92% and anti-TG positivity of
57.3 %. Thyroid evaluation tests including T3, T4, and TSH were abnormal only in patients not receiving L-th
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
Thyroid carcinoma incidence is increasing year after year and ranking second among top ten cancers in Iraq, especially among women, and this increased the requirement for the improvement of the molecular detection accuracy because of its potential role in the early detection. Two single nucleotide polymorphisms (rs1136410, A>G and rs1805414, A>G) in PARP1 gene were found to be associated with thyroid carcinoma risk in several genome wide association studies, therefore, this is a case-control study that was carried out to identify whether these polym
... Show MoreTuberculosis 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 MoreBackground: Alopecia areata(AA) is a common autoimmune disease that causes hair loss without scarring. It occurs as a result of T-helper 1 (Th1) and Th17 cells attacking the anagen hair follicles. Genetic factors play a role in the occurrence of infection, which stimulates the production of pro and anti-inflammatory interleukins. Polymorphisms of IL-37 play a role in autoimmune diseases. However, IL37 single nucleotide polymorphisms(SNP) have not been identified in patients with AA. Therefore, this study aimed to reveal the IL37 gene SNP and its relationship to AA. Methods: Genotyping of IL-37 gene single nucleotide polymorphisms SNPs were detected using sequence-specific primer-polymerase chain reaction (SSP-PCR) method was done following
... Show MoreToll-like receptors (TLRs) play a key role in innate immune response activation against viruses. TLR7, one of the TLRs family, is potentially important in controlling viral infection and the production of vaccines against the virus. The wide spectrum of discrepancy in response to antiviral drugs among different populations which is emerged by some pandemics like COVID-19 might be due to their different TLR7 single nucleotide polymorphisms (SNPs). The present study aimed to investigate the consequences of 401 non-synonymous missense SNPs (nsSNPs) within TLR7 on its protein structure, stability, and function by using specific bioinformatics tools. Seven bioinformatics tools were used to investigate 401
... Show MoreColorectal cancer (CRC) is the most common gastrointestinal malignancy and one of the top ten common cancers worldwide with approximately 2 million cases. There are multiple risk factors that could lead to CRC emergence; of which are genetic polymorphisms. Excision repair cross-complementing group 2 (ERCC2) gene encodes for ERCC2 enzyme which plays a crucial role in maintaining genomic integrity by removing DNA adducts. Several studies suggested that there could be a link between genetic polymorphisms of ERCC2 gene and the risk of CRC development. Hence the present study aims to validate the relationship between the following ERCC2 single nucleotide polymorphisms (rs13181, rs149943175, rs530662943, and rs1799790) and CRC susceptibility. A t
... Show MoreTotal of 46 isolates of Klebsiella pneumoniae were collected from patients attending (Al-Yarmook Hospital and Education Labs / medical city), and isolates were re-identified, depending on morphology and biochemical tests . Disk diffusion method was employed to determine antibiotic susceptibility of forty six isolates by using eleven antibiotics .The results revealed the sensitivity of six isolates (9.3%) to Imipenem and Meropenem . On the other hand the isolates were showed 23.9% resistant against Ciprofloxacin, while some isolates shown higher resistant against several antimicrobial agents such as 65.2%, 69.0% for Amikacin and Cefepime consequently , 71.1%, 71.7 % for Amoxicillin -Clauvulanic acid and Gentamicin and 82.6% against Pipera
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