Background. Helicobacter pylori (H. pylori) is ubiquitous among humans and one of the best-studied examples of an intimate association between bacteria and humans. Phylogeny and Phylogeography of H. pylori strains are known to mirror human migration patterns and reflect significant demographic events in human prehistory. In this study, we analyzed the molecular evolution of H. pylori strains detected from different tribes and regions of Sudan using 16S rRNA gene and the phylogenetic approach. Materials and methods. A total of 75 gastric biopsies were taken from patients who had been referred for endoscopy from different regions of Sudan. The DNA extraction was performed by using the guanidine chloride method. Two sets of primers (universal and specific for H. pylori) were used to amplify the 16S ribosomal gene. Sanger sequencing was applied, and the resulted sequences were matched with the sequences of the National Center for Biotechnology Information (NCBI) nucleotide database. The evolutionary aspects were analyzed using MEGA7 software. Results. Molecular detection of H. pylori has shown that 28 (37.33%) of the patients were positive for H. pylori and no significant differences were found in sociodemographic characteristics, endoscopy series, and H. pylori infection. Nucleotide variations were observed at five nucleotide positions (positions 219, 305, 578, 741, and 763–764), and one insertion mutation (750_InsC_751) was present in sixty-seven percent (7/12) of our strains. These six mutations were detected in regions of the 16S rRNA not closely associated with either tetracycline or tRNA binding sites; 66.67% of them were located in the central domain of 16S rRNA. The phylogenetic analysis of 16S rRNA sequences identified two lineages of H. pylori strains detected from different regions in Sudan. The presence of Sudanese H. pylori strains resembling Hungarian H. pylori strains could reflect the migration of Hungarian people to Sudan or vice versa. Conclusion. This finding emphasizes the significance of studying the phylogeny of H. pylori strains as a discriminatory tool to mirror human migration patterns. In addition, the 16S rRNA gene amplification method was found useful for bacterial identification and phylogeny.
Background Helicobacter pylori (H. pylori) infects nearly half of the world’s population with a variation in incidence among different geographic regions. Genetic variants in the promoter regions of the IL1B gene can affect cytokine expression and creates a condition of hypoacidity which favors the survival and colonization of H. pylori. Therefore, the aim of this study was to characterize the polymorphic sites in the 5′- region [−687_ + 297] of IL1B in H. pylori infection using in silico tools. Results A total of five nucleotide variations were detected in the 5′-regulatory region [−687_ + 297] of IL1B which led to the addition or alteration of transcription factor binding sites (TFBSs) or composite regulatory elements (CEs). Genotyping of IL1B − 31 C > T revealed a significant association between -31 T and susceptibility to H. pylori infection in the studied population (P = 0.0363). Comparative analysis showed conservation rates of IL1B upstream [−368_ + 10] region above 70% in chimpanzee, rhesus monkey, a domesticated dog, cow and rat. Conclusions In H. pylori-infected patients, three detected SNPs (− 338, − 155 and − 31) located in the IL1B promoter were predicted to alter TFBSs and CE, which might affect the gene expression. These in silico predictions provide insight for further experimental in vitro and in vivo studies of the regulation of IL1B expression and its relationship to H. pylori infection. However, the recognition of regulatory motifs by computer algorithms is fundamental for understanding gene expression patterns.
Background: Helicobacter pylori is responsible for gastric cancer in approximately tens of millions of patients. Gastric cancer in Sudan represents one of the top causing death among cancers with about 686 cases per year and a 2.7 % mortality rate. IL-1RN VNTR polymorphism has been reported to increase the risk of gastric cancer. Objective: The purpose of this study was to assess the association of the 86 bp VNTR polymorphism of IL- 1RN gene and the susceptibility to H. pylori infection and gastric cancer in the Sudanese population. Materials and methods: Genomic DNA was extracted from 114 subjects. Of whom 60 had gastritis and duodenitis, 26 had a peptic ulcer, 16 had gastric cancer and 12 had normal gastroscopy findings. H. pylori infection was investigated by specific 16S rRNA. And IL-1RN VNTR polymorphism at intron 2 was genotyped using the PCR method and direct sequencing for random samples. Results: The positive H. pylori infection rate among participants was 47.37%. There is a lack of a significant difference in IL- 1RN genotype with H. pylori infection (p-value=1.0000). The IL-1 RN L/L genotype was significantly more frequent in a patient with benign disorders (gastritis or duodenitis or peptic ulcer), Odd=6.000 (95% CI =1.750-20.57, P=0.0056). While the heterozygote genotype 2/L was associated with an increased risk of gastric cancer with OR = 12.83 (95% CI = 1.261-130.6, P=0.0302). Conclusion: Independently carriage of IL-1RN *2 allele was associated with increased risk of gastric cancer in the Sudanese population. Notwithstanding the relatively small sample size of the study population, our findings show that the host genetic can be a useful tool for identifying high-risk individuals among dyspeptic patients; and also underscore the role played by host genetics in gastric carcinogenesis. To the best of our knowledge, this is the first study in Sudan concerning this issue.
Objective In this study, we analyzed the molecular evolution of Staphylococcus aureus isolates using 16S rRNA gene and phylogenetic analysis to detect the prevalence of S. aureus infections in Sudan. Results Molecular detection of S. aureus has shown that 20 (43.47%) of patients were positive for S. aureus. The phylogenetic tree of 16S rRNA sequences was divided into three lineages of S. aureus isolates detected from wound infections in Sudan. Nucleotides base-pair substitution was appeared at position 249. This mutation do not linked with Macrolides, Lincosamides and Streptogramines b resistant phenotype. Further studies should investigate the effect of that mutation on resistance to other antibiotics.
Background: H. pylori is ubiquitous among humans, and one of the best studied examples of an intimate association between bacteria and humans. Under several diverse socio-demographic factors in Sudan, a continuous increase in the prevalence rate of H. pylori infection has been noticed which represents a major public health challenge. In this study, we analyzed the molecular evolution of H. pylori Strains detected from different ethnic and regions of Sudan using 16S rRNA gene and phylogenetic approach. Materials and methods:A total of 75 gastric biopsies taken from patients who had been referred for endoscopy from different regions of Sudan. The DNA extraction was done by using the guanidine chloride method. Two sets of primers (universal and specific for H. pylori) were used to amplify the 16S ribosomal gene. Sanger sequencing was performed; then Blast these sequences with those available in the NCBI nucleotide database. The evolutionary aspects were analyzed using a MEGA7 software.Result: Molecular detection of H. pylori has shown that 28 (37.33%) of patients were positive for H. pylori. Bivariate analysis has found no significant differences exhibited across sociodemographic, endoscopy series and H. pylori infection. Nucleotide variations were found at five nucleotide positions (positions 219, 305, 578, 741 and 763-764) and one insertion mutation (750_InsC_751) was present in sixty-seven percent (7/12) of our strains. The phylogenetic tree diverged into two lineages. Conclusion:The phylogenetic analysis of 16S rRNA sequences identified two lineages of H. pylori strains detected from different regions in Sudan. Sex mutations were detected in regions of the 16S rRNA not closely associated with either tetracycline or tRNA binding sites. 66.67% of them were located in the central domain of 16S rRNA. Studying the effect of these mutations on the functions of 16S rRNA molecules in protein synthesis and antibiotic resistance is of great importance.
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