Transcriptional Profile of Helicobacter pylori Virulence Genes in Patients with Gastritis and Gastric Cancer

Introduction. Numerous molecular epidemiology studies have been performed about the frequency of Helicobacter pylori virulence genes in patients with H. pylori infection so far. This study was conducted to detect transcriptional profile by cDNA of H. pylori virulence genes in gastric biopsy samples of gastritis and gastric carcinoma patients. Materials and Methods. In a case-control study, based on the prevalence of gastritis and gastric cancer in Sanandaj city during 2018 and 2019, 23 and 11 gastric antral biopsy samples with H. pylori infection were collected from gastritis and gastric carcinoma patients by the consecutive and available sampling method. Pathological characters, including tumor grades and tumor areas for gastric carcinoma biopsy samples prepared from gastric cancer areas, were determined by the pathologist. Total RNA of gastric antral biopsy samples was extracted, and their cDNA was synthesized by TaKaRa kit. H. pylori virulence genes’ cDNA using specific primers and PCR was detected. This study’s results were analyzed by SPSS version 25 and statics chi-square tests for determination of relationship and correlation between cDNAs of H. pylori transcriptional profile and clinical outcomes of H. pylori infection, including gastritis, gastric carcinoma, tumor grades, and tumor area. Results. The positive statistical correlations were observed between transcripts of cagA, cagA-EPIYAC, cagE, and cagY genes and H. pylori infection clinical outcomes ( P < 0.05 ). Conclusion. Detection of the H. pylori virulence genes’ cDNA in gastric biopsy samples can help provide the prognosis of clinical outcomes.

Clinical Endoscopic and Histological Characteristics of Helicobacter Pylori Positive and Negative Armenian Children with Recurrent Abdominal Pain and/or Dyspepsia

BackgroundRecurrent abdominal pain (RAP) and dyspepsia are common complaints in children. These symptoms are often associated with Helicobacter pylori (Hp)infection. Aim of the present study was to prospectively analyze clinical, endoscopic and histological characteristics of Hp+ and Hp- children with RAP and/or dyspepsia. MethodsPatients aged 2-18 years with RAP and/or dyspepsia, referred for upper endoscopy to Arabkir MC from November 2015 to December 2017, were involved in the study. Histology was assessed according to the updated Sydney system. Gastric and duodenal specimens were stained by modified Giemsa staining for Hp infection. One antral biopsy was cultured in Hp selective media. Results150 patients were included into the study: 70.7% Hp+,29.3% Hp-. Nausea and vomiting were significantly more common in Hp+ patients (p<0.05). Gastric nodularity (p=0.02), erosions in the stomach (p=0.056), and duodenal erosions (p=0.019) were more common in Hp+. Chronic active (p=0.027) and non-active gastritis (p=0.002), cumulative findings of metaplasia/dysplasia/atrophy in the stomach (p=0.014) and chronic non-active duodenitis (p=0.016), were significantly more common in Hp+ patients. ConclusionHp infection prevalence is high in Armenian children with dyspepsia and/or RAP. Clinical symptoms, endoscopic findings and histopathological findings were significantly different in Hp+ patients as compared to Hp- patients.

Prospective study to evaluate the number and the location of biopsies in rapid urease test for diagnosis of Helicobacter Pylori

Helicobacter pylori (H. pylori) can cause a wide variety of illnesses such as peptic ulcer disease, gastric adenocarcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma. The diagnosis and eradication of H. pylori are crucial. The diagnosis of H. pylori is usually based on the rapid urease test (RUT) and gastric antral biopsy for histology. The aim of this study is to evaluate the numbers of needed biopsies and their location (antrum/fundus) to obtain optimal result for the diagnosis of H. pylori. Three hundred fifty consecutive patients were recruited, 210 fulfill the inclusion criteria and had nine gastric biopsies for the detection of H. pylori infection: two antral for the first RUT (RUT1), one antral and one fundic for the second (RUT2), one antral for the third (RUT3) and two antral with two fundic for histology (HES, Giemsa, PAS). The reading of the 3 types of RUT was performed at 1 hour, 3 hours and 24 hours and biopsies were read by two experienced pathologists not informed about the result of RUT. Results of RUT were considered positive if H. pylori was found on histology of at least one biopsy. The RUT1 at 1h, 3h and 24h has a sensitivity of 72%, 82% and 89% and a specificity of 100%, 99% and 87% respectively. The positive predictive value (PPV) was 100%, 99% and 85% respectively and the negative predictive value (NPV) of 81%, 87% and 90%. The RUT2 at 1h, 3h and 24h, respectively, had a sensitivity of 86%, 87% and 91% and a specificity of 99%, 97% and 90%. The PPV was 99%, 96% and 88% and NPV of 89%, 90%, 94%. The RUT3 at 1h, 3h and 24h, respectively, had a sensitivity of 70%, 74% and 84% and a specificity of 99%, 99% and 94%. The PPV was 99%, 99% and 92% and NPV of 79%, 81% and 87%. The best sensitivity and specificity were obtained for RUT1 read at 3h, for RUT2 read 1h and 3h, and the RUT3 read at 24h.This study demonstrates that the best sensitivity and specificity of rapid test for urease is obtained when fundic plus antral biopsy specimens are used with a reading time at 3 hours.

Helicobacter pyloriIs Not Eradicated after Triple Therapy: A Nested PCR Based Study

Detection ofHelicobacter pyloriafter triple therapy is usually carried out by either rapid urease test (RUT), urea breath test (UBT), histology, bacterial isolation, and single round PCR or serological tests. In this study, antral biopsy specimens from 25 patients were tested forH. pyloriby RUT, culture, histology, and nested PCR in their antral biopsy specimens before and after treatment. Three genes, namely, heat shock protein (hsp60), phosphoglucosamine mutase (ureC), and flagellar export ATP synthase (fliI) ofH. pyloriwere targeted. Of the 25 antral biopsy specimens, the RUT, culture, histology, and nested PCR positivity dropped from 81.8% to 12%, 31% to 0%, 100 to 84%, and 100% to 92%, respectively, before and after therapy. Further,hsp60specific amplicons from 23 out of 25 patients gave identical restriction pattern, while 6fliIand 1ureCspecific amplicon produced different restriction pattern. Furthermore, variations infliIgene sequences inH. pyloriafter treatment were also confirmed by sequencing and comparedin silico. Nested PCR based detection ofH. pyloriis more sensitive method to detectH. pyloriafter therapy than culture, RUT, and histology. Further, this study suggests thatH. pyloriis not eradicated completely after triple therapy.

Performance of RoutineHelicobacter pyloriInvasive Tests in Patients with Dyspepsia

Background. This study was designed to compare the accuracy of three different invasive methods for the detection ofHelicobacter pylori (H. pylori)infection in patients with dyspepsia. These tests included culture, histology, and the rapid urease test (CLO test).Methods.H. pyloriinfection was diagnosed prospectively in 246 untreated dyspeptic patients who underwent upper gastrointestinal endoscopy. The gold standard forH. pyloriinfection was based on a positive culture or both a positive histological examination and a CLO test.Results.H. pyloriwas diagnosed in 33.3% of the patients. The sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy were as follows: histology from the antrum (95.12; 95.12; 90.7; 97.5; 95.12%); histology from the antrum and corpus (95.12; 95.12; 90.7; 97.5; 95.12%); histology from the corpus (76.83; 96.95; 92.65; 89.33; 90.24%); culture (91.46; 100; 100; 95.91; 97.15%); a CLO test from the antrum and corpus (85.59; 100; 100; 93.71; 95.52%); a CLO test from the antrum (64.63; 100; 100; 84.97; 88.21%); a CLO test from the corpus (69.51; 100; 100; 96.77; 89.83%), respectively.Conclusions. Antral biopsy histology and culture are the best methods for the diagnosis ofH. pyloriinfection in our cohort of patients with dyspepsia.

Fluorescence in situ hybridization biomarkers of gastric cancer risk in intestinalized Helicobacter pylori pangastritis.

17 Background: H. pylori (HP) intestinalized pangastritis predisposes to gastric adenocarcinoma. Biomarkers to identify the less than 10% subset of chronic gastritis patients who will develop gastric cancer are greatly needed to focus surveillance efforts onto those patients most likely to benefit. Investigations in ulcerative colitis and Barrett's esophagus reveal that chromosomal alterations in far distant and nondysplastic mucosa are powerful cancer biomarkers. We hypothesized a similar situation in gastric tumorigenesis. Methods: The full morphologic spectrum of gastric neoplasia was examined using flow-sorted gastric epithelial cells from 10 gastric cancer resections versus 20 normal gastric control patients. Then we addressed whether we could distinguish the subset of HP intestinalized pangastritis patients who develop cancer (progressors), from those who remain neoplasia-free (nonprogressors). A single nondysplastic intestinalized gastric antral biopsy collected using chromoendoscopy on each of 41 Korean progressors with gastric cancer and 30 nonprogressors without dysplasia. Interphase FISH on whole epithelial cell preparations using 4q32-q33, 10q22.1, 18q21.2 and 19p11-q11 chromosomal arm probes was performed. Results: Increasing chromosomal alterations occurred throughout the morphologic spectrum of gastric neoplasia. Significantly increased FISH alterations for chromosomes 4q32-q33 (p = 0.016) and 10q22.1 (p = 0.028) occurred as early in the neoplastic spectrum as nondysplastic distant intestinal metaplasia. Subsequent nondysplastic antral biopsy testing of progressors versus nonprogressors revealed parallel FISH results for 4q32-q33 (p = 0.039) and 10q22.1 (p = 0.023), and 19p11-q11 (p = 0.019). Receiver operator characteristic analysis for these 3 combined probes demonstrated a sensitivity of 0.53 and specificity of 0.92 with optimal choice of thresholds. Conclusions: Interphase FISH for 4q32-q33, 10q22.1, and 19p11-q11 chromosomal probes on nondysplastic intestinalized antral mucosa shows promise as a biomarker of gastric cancer in multifocal HP intestinalized pangastritis. No significant financial relationships to disclose.

Detection of Helicobacter pylori in gastric biopsy and resection specimens

Aim. To compare the sensitivity of detecting H. pylori in gastric biopsy and resection specimens using modified Giemsa stain and immunohistochemistry, using a commercially available anti-H. pylori antibody (Dako, Denmark). Methods. Gastric antral biopsy specimens showing chronic gastritis (28 cases) together with tissue blocks from gastrectomy specimens for duodenal ulcer (2 cases) were stained with modified Giemsa and immunoenzymatic alkaline phosphatase - anti-alkaline phosphatase (APAAP) method, and were carefully examined for the presence of H. pylori. Results. Using a modified Giemsa stain, the spiral shaped bacteria of H. pylori stained blue, were attached to the brush border of the gastric foveolar epithelial cells. However, the specificity of modified Giemsa stain depended on the morphological appearance of H. pylori. The specificity of immunostaining permitted detection of low numbers or even single organisms. In all cases bacteria were more prominent and easier to detect in immunostained preparations. H. pylori was identified in 22 (73.3%) of 30 sections stained with modified Giemsa stain, but it could be identified with greater frequency in sections stained with APAAP, in 27 (90%) of 30 sections. Conclusion. Immunohistochemical identification of H. pylori was better than Giemsa stain for detecting that organism.