BackgroundThe aim of this study was to identify the common H. pylori virulence genes among dyspeptic Southwestern Saudi patients and their association with clinical outcomes and histopathological findings to help practitioners and researchers in the region for better management of infections caused by such bacteria.MethodsFour hundred two gastric biopsy specimens were analyzed using histopathological examination and real time-PCR. The positive 187 specimens by RT-PCR were genotyped using PCR targeting cagA, vacA and iceA genes.ResultsOne hundred twenty-eight gastric biopsy specimens were positive in genotyping PCRs. The cagA, vacA, iceA1 and iceA2 genes were detected in rates of 49.2% (63/128), 100%(128/128), 42.2% (54/128), 32.8% (42/128), respectively. The vacA s1as1bm2 subtype was the highest 23.4% (30/128), followed by m2 and s1a1b subtypes which were equally detected [16.4% (21/128) for each]. The iceA genes were significantly associated with gastritis and gastric ulcer. Overall, vacA genotypes were significantly associated with gastritis, gastric and duodenal ulcers. The vacA subtypes: s1as1bm2, s1a1b and s2 m2 showed chronic active gastritis in percentages of 90.0, 81, and 84.2%, respectively. All vacA mixed genotypes showed chronic active gastritis.ConclusionsH. pylori virulence genes are highly prevalent and diverse among patients with dyspepsia in Southwestern region of Saudi Arabia. The iceA genes and the different vacA subtypes are significantly associated with the clinical outcomes and histopathological changes especially chronic active gastritis.
BackgroundHelicobacter pylori (H. pylori) is a major cause of peptic ulcer disease (PUD) and chronic active gastritis that may progress to gastric cancer. Globally, it has been estimated that 50% or more of the world’s population is infected by H. pylori, making it the most widespread infection across the globe.ObjectivesTo determine the prevalence of H. pylori infection and to identify factors associated with H. pylori infection in Saudi patients presenting with dyspepsia.MethodsIn this prospective cross-sectional study, a total of 404 gastric biopsies were endoscopically obtained from 404 patients with dyspepsia from September 2014 to April 2016 (Jazan Province, Saudi Arabia). The specimens were analyzed using the real-time polymerase chain reaction (PCR). The data was examined using descriptive statistics as well as determining the prevalence, and employing Chi square and Fisher exact test. A p-value of ≤0.05 was considered statistically significant in examining the research hypotheses.ResultsThe overall prevalence of H. pylori in Jazan Province was 46.5% (95% CI: 41.7–51.4) and the prevalence was lower among those > 55 years old. Prevalence was higher among urban (50.0%; 95% CI: 43.1–56.8) versus rural (42.1%; 95% CI: 35.1–49.3), but with no significant difference. Prevalence did not show significant difference among different Body Mass Index (BMI) categories, ranging from 40.2% to 47.7%. The prevalence of H. pylori in females was 47.1% (95% CI: 40.4–53.9) versus 45.6% (95% CI: 38.7–52.6) in males. Histopathology findings were associated with H. pylori infection with prevalence of 58.1% among patients with chronic active gastritis, compared to 24.1% and 34.8% among mild and chronic gastritis, respectively.ConclusionOur results indicate that there is a high prevalence of H. pylori among Saudi patients with dyspepsia. Prevalence of H. pylori was high in ages below 55 years. Chronic active gastritis was significantly associated with H. pylori infection. In depth studies are needed to determine associated factors with of H pylori infection in the region
Gastric Helicobacter pylori infection is diagnosed based on histopathological evaluation of gastric mucosal biopsies, urease test, urea breath test, H. pylori culturing, or direct detection using polymerase chain reaction (PCR). This study aimed to evaluate the efficacy of immunohistochemical (IHC) staining in detecting H. pylori in gastric biopsies from patients with chronic gastritis and minimal or atypical infection. Gastric biopsies from 50 patients with chronic gastritis were subjected to routine haematoxylin and eosin (H&E), modified Giemsa, and IHC staining. The results of staining were compared with those of quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis identified 32 (64%) H. pylori-positive cases, whereas IHC, H&E, and modified Giemsa staining identified 29 (58%), 27 (54%), and 21 (42%) positive cases. The sensitivity of IHC staining (87.50%) was higher than that of H&E (59.38%) and modified Giemsa (43.75%) staining. The specificity of H&E, modified Giemsa, and IHC staining was 55.56%, 61.11%, and 94.44%, respectively. IHC staining exhibited the highest diagnostic accuracy (90%), followed by H&E (58%) and modified Giemsa (50%) staining. Active gastritis, intestinal metaplasia, and lymphoid follicles were detected in 32 (64%), 4 (8%), and 22 (44%) cases, respectively, and all of these cases were H. pylori positive. In contrast to routine H&E and modified Giemsa staining, IHC allows for the accurate H. pylori detection in cases with minimal or atypical infection. Moreover, IHC can be an alternative diagnostic method to qRT-PCR for detection of H. pylori in such cases.
Background Gastric Helicobacter pylori infection is diagnosed based on histopathological evaluation of gastric mucosal biopsies, urease test, urea breath test, H. pylori culturing, or direct detection using polymerase chain reaction (PCR). This study aimed to evaluate the efficacy of immunohistochemical (IHC) staining in detecting H. pylori in gastric biopsies from dyspeptic patients with minimal and/or atypical infection. Gastric biopsies from 50 patients with chronic gastritis were subjected to routine haematoxylin and eosin (H&E), modified Giemsa, and IHC staining. The results of staining were compared with those of quantitative real-time PCR (qRT-PCR). Results The qRT-PCR analysis identified 32 (64%) H. pylori-positive cases, whereas IHC, H&E, and modified Giemsa staining identified 29 (58%), 27 (54%), and 21 (42%) positive cases. The false-positive rates of H&E and modified Giemsa staining were 16% and 14%, respectively. The sensitivity of IHC staining (87.50%) was higher than that of H&E (59.38%) and modified Giemsa (43.75%) staining. The specificity of H&E, modified Giemsa, and IHC staining was 55.56%, 61.11%, and 94.44%, respectively. IHC staining exhibited the highest diagnostic accuracy (90%), followed by H&E (58%) and modified Giemsa (50%) staining. Active gastritis, intestinal metaplasia, and lymphoid follicles were detected in 32 (64%), 4 (8%), and 22 (44%) cases, respectively, and all of these cases were H. pylori positive. Conclusions H. pylori can be detected using routine H&E or modified Giemsa staining. However, the high sensitivity, specificity, and diagnostic accuracy of IHC staining minimise the false-positive/negative results and enable H. pylori detection in cases with minimal or atypical infection. Moreover, IHC can be an alternative diagnostic method to qRT-PCR for detection of H. pylori in such cases.
Context.—The endometrium is an intrinsically dynamic tissue with great capability for regeneration and proliferation; consequently, there is some overlap between features seen in benign, premalignant, and malignant lesions. This leads to marked intrabiopsy, interbiopsy, and interobserver variability. Objective.—We studied the specificity and sensitivity of computerized image analysis of molecular markers to evaluate its potential use as a diagnostic tool. Design.—Specimens from 100 patients were examined and the following histologic diagnoses were assigned: proliferative endometrium (n = 10), secretory endometrium (n = 10), endometrial hyperplasia (n = 40; 30 with no atypia, 10 with atypia), and carcinoma (n = 40; 20 endometrioid, 10 serous, and 10 clear cell). All cases were evaluated immunohistochemically for p53 and proliferating cell nuclear antigen (PCNA) expression. Computerized image analysis was performed with a CAS 200 digital analyzer. Results.—Expression of p53 was found only in carcinomas (65%) and endometrial hyperplasia with atypia (30%). Expression of p53 was higher in the poor prognostic categories (serous carcinoma and clear cell carcinoma) than in endometrioid carcinoma. In endometrioid carcinoma, p53 expression correlated with grade. Proliferating cell nuclear antigen showed a similar pattern of results to p53 in the various carcinoma subtypes and endometrioid carcinoma grades. Endometrial hyperplasia PCNA values were the lowest among all the groups. Both carcinomas and proliferative endometrium showed higher glandular and stromal PCNA values, significantly different from endometrial hyperplasia with atypia. In proliferative endometrium, stromal PCNA was the highest among all of the groups. The p53 and PCNA results correlated with each other for carcinoma. Conclusions.—Computerized image analysis correlates well with the established morphologic groups of endometrial pathology and yields results consistent with previous studies. Owing to its higher degree of sensitivity, computerized image analysis is of potential use in cases of diagnostic dilemmas and can help objectively allocate the case in the correct category (eg, proliferative endometrium vs endometrial hyperplasia, endometrial hyperplasia with atypia vs endometrioid carcinoma). It is particularly useful in the evaluation of stromal changes.
Background: Conventional polymerase chain reaction (PCR)-based methods play a major role in the direct detection of H. pylori in clinical specimens, with time-saving as compared to culture-based methods. However, specificity and sensitivity vary among different varieties of these PCRs, which consequently could affect the accuracy of diagnosis of H. pylori infection. The study aimed to evaluate the utility of ureC (glmM) and SSA conventional PCR methods for rapid direct detection of H. pylori by comparing them with rpoB-based quantitative real-time PCR. Methods: A total of 402 non-repeated gastric biopsy specimens were subjected to DNA extraction followed by conventional ureC (glmM) and SSA PCR, and rpoB-based quantitative real-time PCR, which was used as the gold standard. Results: H. pylori was detected in 119 (29.6%), 126 (31.34%), and 187 (46.5%) of the tested specimens using ureC (glmM) PCR, SSA PCR, and real-time quantitative PCR, respectively. The specificity of the SSA PCR was higher than that of ureC (glmM) PCR (99.5% and 98.6%, respectively). The SSA PCR was more sensitive than the ureC (glmM), (66.8% and 62%, respectively). The diagnostic accuracy of SSA PCR (84.33%) was higher than that of ureC (glmM) PCR (81.59%). Conclusion: Overall, SSA PCR is more specific, sensitive, and diagnostically accurate than ureC (glmM) PCR, giving the SSA PCR assay superiority as a simple, rapid, and accurate diagnostic tool for direct detection of H. pylori in gastric tissue specimens.
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