Background: Effective drug regimens for the treatment of hepatitis B virus (HBV) infections are essential to achieve the World Health Organisation commitment to eliminate viral hepatitis by 2030. Lamivudine (3TC) is widely used in countries with high levels of chronic HBV, however resistance has been shown to occur in up to 50% of individuals receiving continuous monotherapy for 4 years. Telbivudine (LdT) is now more commonly used in place of lamivudine but is ineffective against 3TC-resistant HBV. Genotyping and identification of resistance-associated substitutions (RAS) is not practical in many locations.Objectives: A novel assay was designed to enable HBV genotyping and characterisation of resistance mutations directly from serum samples stored on filter paper, using Sanger and MinION sequencing.
Study design:The assay was applied to a cohort of 30 samples stored on filter paper for several years with HBV viral loads ranging from 8.2x10 8 to 635 IU/mL. A set of 6 high-titre samples were used in a proof-of-principle study using the MinION sequencer.
Results:The assay allowed determination of HBV genotype and elucidation of RAS down to 600 IU/mL using a 550bp amplicon. Sequencing of a 1.2kb amplicon using a MinION sequencer gave results consistent with Sanger and allowed the identification of minor populations of variants.
Conclusions:We present two methods for reliable HBV sequencing and RAS identification using methods suitable for resource-limited environments. This is the first demonstration of extraction-free DNA sequencing direct from DSS using MinION and these workflows are adaptable to the investigation of other DNA viruses.
Objectives
Early-onset myocardial infarction constitutes nearly one third of cases of myocardial infarction among Iraqis, which is rather higher than the proportions reported in many Western countries. Thus this study was initiated to investigate the role of some genetic polymorphisms, as well as acquired risk factors in this condition.
Results
A total of 102 Iraqi patients with first myocardial infarction aged 50 years, and 77 matched controls were enrolled. The DNAs of participants were screened for nine polymorphisms, namely: β-Fibrinogen (− 455G > A), Factor XIII (V34L), Plasminogen Activator inhibitor-1 (PAI-1, 4G/5G), Human Platelet Antigen-1 (HPA1a/b), 5,10-Methylenetetrahydrofolate Reductase MTHFR (C677T) and MTHFR (A1298C), Angiotensin-Converting Enzyme (ACE) 287 bp insertion/deletion (I/D), Apolipoprotein-B (ApoB: R3500Q), and Apolipoprotein-E (Apo E: E2/E3/E4), using PCR and reverse hybridization technique. Among traditional risk factors, univariate analysis revealed that smoking (OR 2.86 [95%CI 1.53–5.34]), hyperlipidemia (OR 5.23 [95%CI 2.66–10.29]), and diabetes mellitus (OR 4.05 [95% CI 1.57–10.41]) were significantly higher among patients compared to controls (P<0.001, <0.001 and 0.002 respectively), while none of the nine genetic polymorphisms reached significance. Multivariate Logistic regression, however, revealed that only smoking and hyperlipidemia retained significance (P of < 0.001 each). The need to initiate further studies on larger cohorts is paramount to understand the higher than expected frequency of early-onset myocardial infarction in our population.
Background: Smoking is the foremost public health problem affecting the world and it has a crucial implication in causing many common diseases due to Helicobacter pylori infection which is globally distributed. Smoking is considered a critical risk factor that accelerates infection with this bacterium.Objectives: The study's goal was to find out how common Helicobacter pylori infections were among male cigarette and nargileh smokers. Materials and methods: A case-control study was performed between August and December 2021in Erbil Teaching Hospital in Erbil City, Kurdistan Region, Iraq. Blood samples were collected and used for detection of anti-Helicobacter pylori IgG Ab for 80 males who were smokers and 80 who were non-smoker. Results: The prevalence of Helicobacter pylori positivity was 64.9% in smokers and 45.5% in nonsmokers (P-Value = 0.03). The highest percentage (54.1%) was found in the young age group (25-34) years (P-Value = 0.05), and 89.2% of Helicobacter pylori-positive individuals exhibited stomach symptoms (P-Value = 0.01). Fifty percent of Helicobacter pylori-infected individuals were nargileh smokers.
Conclusion:The prevalence of Helicobacter pylori showed significant value in nargileh smoking males. Therefore, smoking was a key factor in the seroprevalence of Helicobacter pylori and had a substantial impact on it.
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