Abstract
Background: The continuous rise in the number of patients suffering from Helicobacter pylori is probably due to the changes in modern life. Nowadays, patients suffering from gastrointestinal problems are diagnosed through invasive and non-invasive techniques. The choice of a diagnostic test is influenced by factors such as the tests' sensitivity and specificity, the clinical conditions, and the cost-effectiveness of the testing strategy. This study aimed to compare molecular detection methods of H. pylori by polymerase chain reaction (PCR) targeting the 16S rRNA, ureA and glmM genes with an invasive histopathological technique. Methods: 290 gastric biopsies were collected using gastrointestinal endoscopy from patients with gastritis symptoms in different hospitals in Khartoum state. Two gastric biopsies were collected from each patient for PCR and histopathology. Results: A total of 103 (35.5%) samples were positive by histopathological examination, 88 (30.3%) by 16S rRNA, 39 (13.4%) by glmM gene, and 56 (19.3%) by ureA gene. The highest sensitivity was observed in 16S rRNA (46.6%), followed by glmM (24.3%) and ureA (23.3%). While the best specificity was observed in glmM gene (92.5%), followed by ureA (82.3%) and 16S rRNA (78.6%). Conclusion: PCR test targeting the 16S rRNA gene exhibited the best results for molecular detection of H. pylori compared to other genes.
Highlights
The sensitivity of any of those techniques in detecting H. pylori relays on how the density of the bacterial cells within the specimens taken by biopsy (recent use of disease-related medications, antibiotics and protonpump inhibitors (PPI) can reduce the density of the cells), pathologist expertise, the type and quality of the stain used for detection purposes.[10]
H. pylori were clearly detected in positive samples as curved bacilli on the surface of the gastric epithelial cells; the bacteria appear as light bluish rods in H and E slides with varying sizes (3–6 μ) on the luminal surface of mucosal cells
Salman et al reported that 115/210 (54.7%) samples were positive for H. pylori via histopathology, 57 (62.6%) of positive H. pylori samples were observed in patients with chronic gastritis, 11 (50%) with adenocarcinoma and 31 (44.2%) with superficial gastritis, while only one H. pylori-positive out of 5 cases observed in atrophy gastritis patient.[29]
Summary
Helicobacter pylori (H. pylori) is a Gram-negative, microaerophilic, spiral, and motile bacterium that colonizes the human gastric mucosa.[1,2] It has been associated with the development of various clinical disorders of the upper gastrointestinal tract, such as aseptic ulcers, chronic gastritis, gastric adenocarcinoma and gastric mucosa-associated lymphoid tissue (MALT) lymphoma, which is classified as type I cancer-causing agent by the World Health Organization (WHO).[3,4,5] Its distribution is worldwide and affects more than 90% of the world population, but it is more common in developing countries with the highest prevalence found in Africa,[6,7] probably due to the possible transmission through the fecal-oral route and the unsafe sanitation conditions in these countries.[1,8] Clinically, a variety of various invasive techniques (requiring endoscopy and biopsy which include, culture, histological examination, and rapid urease test, CLO (Campylobacter like organism) test, smear examination, and molecular studies) or noninvasive techniques (including serology, respiratory urea breath test, or the detection of fecal antigen) are often performed to detect H. pylori infection.[9,10,11] The sensitivity of any of those techniques in detecting H. pylori relays on how the density of the bacterial cells within the specimens taken by biopsy (recent use of disease-related medications, antibiotics and protonpump inhibitors (PPI) can reduce the density of the cells), pathologist expertise, the type and quality of the stain used for detection purposes.[10]. This study aimed to compare molecular detection methods of H. pylori by polymerase chain reaction (PCR) targeting the 16S rRNA, ureA and glmM genes with an invasive histopathological technique. Methods: 290 gastric biopsies were collected using gastrointestinal endoscopy from patients with gastritis symptoms in different hospitals in Khartoum state. Two gastric biopsies were collected from each patient for PCR and histopathology. The highest sensitivity was observed in 16S rRNA (46.6%), followed by glmM (24.3%) and ureA (23.3%). While the best specificity was observed in glmM gene (92.5%), version 1
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