scholarly journals Detection of Helicobacter pylori Through Histochemistry & Immunofluorescent Staining in Biopsies of Patients with Chronic Gastritis

Author(s):  
Asia Jahanzeb ◽  
Abdul Hannan Nagi Nagi ◽  
Ayesha Suleman

Introduction: Helicobacter pylori (H.pylori) is a helix shaped gram negative rod which is usually associated with chronic gastritis and also a major cause of other astroduodenal diseases as well. Aims & Objectives: The present study used histochemical and immunofluorescent stains on formalin fixed paraffin embedded human gastric biopsies for detection of H.pylori. Comparison was also done to evaluate best staining method. Place and duration of study: This study was accomplished in about one year. Sampling of gastric biopsies and rapid urease test were executed at the endoscopy suite of Lahore General Hospital whereas the histopathological examination and immunofluorescent staining were done in University of Health Sciences, Lahore. Material & Methods: Thirty patients (n=30cases) were included in the study following inclusion criteria. Diagnostic upper GI endoscopy was carried out in all cases. Five gastric biopsies were taken from each patient/case (total N=150 Biopsies) according to the Updated Sydney System. Rapid urease test was performed at the site of endoscopy. Biopsies fixed in 10% formalin were brought to the concerned department where they were assigned a specific laboratory number then processed and stained. Results: Endoscopic examination revealed chronic gastritis and rapid urease tests were positive. All cases (n=30) were positive for H.pylori on histopathology. The calculated sensitivity and specificity of H&E, Giemsa, Modified McMullen’s stain and Immunofluorescent method in present study were 71% and 100%,83% and 100%, 82% and 100%, 90% and 100% respectively. Conclusion: Special stains makes H.pylori identification easier in tissue sections. However, immunofluorescent test is the most sensitive and specific method as compared to histochemical stains.

2020 ◽  
Vol 29 (3) ◽  
pp. 59-64
Author(s):  
Hanaa M. El Maghraby ◽  
Samar Mohaseb

Background: Metronidazole is one of the antimicrobial drugs that can be used in combination with other drugs for eradication of Helicobacter pylori (H. pylori).Unfortunately, metronidazole resistance in H. plori is an increasing health problem which may be attributed to inactivation of many genes as rdx A gene. Objective: To determine the frequency of rdx A deletion mutation in H. pylori detected in infected patients attending at the Gastroenterology Unit, Zagazig University Hospitals. Methodology: Two gastric biopsies were taken from each enrolled patient by endoscopy. H.pylori detection was done by rapid urease test and polymerase chain reaction (PCR) amplification of 16S rRNA gene. Deletion mutation in rdx A gene was detected by conventional PCR. Results: Out of 134 doubled gastric biopsies obtained from 134 patients, 52.2% were positive for H. pylori. Epigastric pain, vomiting and gastritis were significantly associated with detection of H. pylori infection (p˂ 0.05). Deletion mutation of rdx A gene was detected in 28.6% of H. pylori positive specimens obtained from infected patients. Conclusion: Deletion mutation of rdx A gene is a frequent determinant of rdx A inactivation conferring metronidazole resistance among H. pylori.


2019 ◽  
Vol 8 (7) ◽  
pp. 1071 ◽  
Author(s):  
Izabela Korona-Glowniak ◽  
Halina Cichoz-Lach ◽  
Radoslaw Siwiec ◽  
Sylwia Andrzejczuk ◽  
Andrzej Glowniak ◽  
...  

The aim of this study was to investigate genetic diversity of Helicobacter pylori virulence markers to predict clinical outcome as well as to determine an antibiotic susceptibility of H. pylori strains in Poland. Gastric biopsies from 132 patients with gastrointestinal disorders were tested for presence of H. pylori with the use of rapid urease test, microbial culture, and polymerase chain reaction (PCR) detection. The genetic diversity of 62 H. pylori positive samples was evaluated by detection of cagA and PCR-typing of vacA and iceA virulence-associated genes. Most common H. pylori genotypes were cagA(+)vacAs1m2 (27.4%) and cagA(−)vacAs2m2 (24.2%). In logistic regression analysis, we recognized the subsequent significant associations: gastritis with ureC, i.e., H. pylori infection (p = 0.006), BMI index (p = 0.032); and negatively with iceA1 (p = 0.049) and peptic ulcer with cagA (p = 0.018). Thirty-five H. pylori strains were cultured and tested by E-test method showing that 49% of strains were resistant to at least one of the tested antibiotics. This is the first study that reports the high incidence and diversity of allelic combination of virulence genes in gastroduodenitis patients in Poland. Genotyping of H. pylori strains confirmed the involvement of cagA gene and vacAs1m1 genotype in development and severity of gastric disorder.


2020 ◽  
Vol 14 (03) ◽  
pp. 298-303 ◽  
Author(s):  
Tan Attila ◽  
Mujdat Zeybel ◽  
Yesim Esen Yigit ◽  
Bulent Baran ◽  
Emel Ahishali ◽  
...  

Introduction: Socioeconomic factors play an important role in the prevalence of Helicobacter pylori (HP) infection. The aim of this study is to investigate HP prevalence among symptomatic patients in the upper socioeconomic segment of the population undergoing gastroscopy in an endemic urban region. Methodology: Over a 12-month period, data were collected from the first consecutive 1000 patients (500 from university hospital, 500 from community hospital) who had gastroscopy and HP evaluation. Results: Overall, 211/1000 patients (21.1 %) were found to have HP in gastric biopsies. The specificity, sensitivity, positive predictive value, negative predictive value and diagnostic accuracy of rapid urease test were 87.5%, 99.7%, 99%, 96.5%, and 96.9% respectively. Atrophic gastritis, gastric and duodenal ulcers were significantly more common in HP positive patients. Age based distribution of HP prevalence: > 6 decades (15.5%), 3rd-5th decades (26.1%), < 3rd decades (10.4%). Conclusion: In an HP endemic country, the prevalence of HP infection among symptomatic patients belonging to the upper socioeconomic segment of the population appears to be markedly lower. The lowest prevalence in young patients is expected to result in future decrease in HP prevalence.


2016 ◽  
pp. 88-93
Author(s):  
Thi Hoai Thai ◽  
Van Huy Tran

Background: H. pylori eradication still remains a challenge to clinicians, especially with the increasing antibiotic-resistant H. pylori. Concomitant therapy showed effective, even in some multiresistant population, but data in Vietnam is still very limited. The study ''Study of Helicobacter pylori eradication with RACM regimen in chronic gastritis patients at Da Nang Hospital from 1/4/2014 to 30/6/2015, is aimed at: (1) Evaluating the results of Helicobacter pylori eradication of Amoxicillin-Clarithromycin-Rabeprazole-Metronidazole therapy for 14 days.(2) Assessing some side effects of this regimen.Method: prospective, consisting of 83 patients examined and treated in Danang hospital from1/ 4/2014 to 30/6/2015, H.pylori was tested by rapid Urease test; H.pylori positive patients received RACM for 14 days. Results: H.pylori eradication rate was 83.1%. H. pylori eradication rates in different locations: antrum 63.8%, higher than corpus (17.4%), antrum and corpus (18.8%), with statistical significance at p<0.05. Common side effects was nausea (27.7%), diarrhea (19.3%). Abdominal pain, lightheadedness, dizziness, insomnia, headache account for low percentage: 8%; 6%; 3,6% and 2.4% respectively. Conclusion: The effect of 14 day RACM regimen for H. pylori eradication was 83.1%, common side effects are nausea (27.7%), diarrhea (19.3%). Key words: chronic gastritis;H. pylori; eradication of H. pylori(ITT); RACM regimen.


2016 ◽  
pp. 149-158
Author(s):  
Ngoc Quy Hue Dang ◽  
Van Huy Tran ◽  
Thanh Hai Nguyen

Background: there has not been yet any research on the effectiveness of H. pylori eradication of bismuth-containing quadruple regimen on chronic gastritis patients in our country. Objective: to evaluate H. pylori eradication rate of bismuth-containing quadruple regimen according to intention to treat (ITT), per protocol (PP) analysis, the rate of side effects and medication compliance. Subjects and Methods: from March 2014 to January 2016 we used bismuth-containing quadruple regimen (EBMT) 10 days for H. pylori eradication therapy for 166 chronic gastritis patients diagnosed based on clinical, endoscopic, rapid urease test, histology and culture. Patients were evaluated side effects and medication compliance at the end of treatment (day 11-14). To assess the eradication, repeating endoscopy with both rapid urease test and histological examination were performed at 4 to 8 weeks after stopping treatment course. Results: H. pylori eradication rates on ITT and PP analysis overall, for naïve patients, after one and more two eradication failures were respectively: 80.72-89.33%, 79.51-90.65%, 91.67-91.67% and 75.00-78.95%. Medication adherence rate was 96.99%. The rates of patients experiencing moderate, severe and very severe side effects were: 19.88%, 0.60% and 1.81%. Conclusion: the EBMT 10-day regimen attained high eradication rates in chronic gastritis patients with rare serious side effects and the high compliance rate. We should apply bismuth-containing quadruple regimen in H. pylori eradication therapy for naïve patients or after one eradication failure. Key words: bismuth-containing quadruple regimen, EBMT, eradication, chronic Helicobacter pylori gastritis


2017 ◽  
Vol 25 (4) ◽  
pp. 214-20 ◽  
Author(s):  
Mohammed S. Alhussaini

Background: Helicobacter pylori is an important gastrointestinal pathogen associated with gastritis, peptic ulcers, and an increased risk of gastric carcinoma. The present study was carried out to determine the relationship between this organism with different gastrointestinal ailments.Methods: 150 outpatients referrals to Saudi Arabian Medical City, Riyadh, Kingdom of Saudi Arabia was recruited in January to June 2015. Each patient was subjected to endoscopic examination. Biopsy specimens were taken from the stomach for rapid urease test and culture. Suspected H. pylori colonies were subjected to colony morphology identification, microscopical examination and biochemical reactions. The samples were also subjected to PCR to detect ureA subunit of urease gene.Results: The endoscopic examination of patients revealed normal, gastric ulcer, duodenal ulcer, gastritis, and gastric cancer with a rate of 20.7%, 20%, 24%, 33.3%, and 2%, respectively. Direct smear exam revealed that 52% of patients were H. pylori positive while culture and rapid urease test showed a prevalence of 71.33%. Fifty four biopsies (36%) were urease positive after 1 hour at room temperature, 39 (62%) after 1 hour incubation at 37°C and 14 (71.33%) after 24 hours incubation. Isolated H. pylori showed that they were catalase, oxidase, and urease positive. PCR results showed 411-bp fragment, which is indicative for the ureA subunit of urease gene.Conclusion: The prevalence of H. pylori infection was high among tested population. Strong association between H. pylori and duodenal ulcer was noticed. A 411-bp fragment indicative of the ureA subunit of urease gene was detected in all the tested isolates.


2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Antoine Abou Rached ◽  
Jowana Saba ◽  
Cesar Yaghi ◽  
Joyce Sanyour ◽  
Ahmad El Hajjar ◽  
...  

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.


2021 ◽  
Vol 65 (3) ◽  
Author(s):  
Mohammed Akeel ◽  
Ahmed Elhafey ◽  
Atef Shehata ◽  
Erwa Elmakki ◽  
Thanaa Aboshouk ◽  
...  

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.


2018 ◽  
Vol 5 (4) ◽  
pp. 1315
Author(s):  
Mrutyunjay I. Uppin ◽  
Kapildev K. Hannurkar

Background: Prevalence of Helicobacter pylori (H. pylori) emerges throughout the world and instigates peptic ulcer disease (PUD). The study was conducted with the aim to determine the prevalence of H. pylori in patients with PUD undergoing upper gastrointestinal endoscopy.Methods: This prospective study was conducted on 150 cases of PUD from August 2009 to February 2011. Endoscopy was done in all cases. Biopsy was done and sent for histopathological examination and rapid urease test for confirmation of presence of H. pylori.Results: Out of 150 patients with mean age of 45.76 years, 109 patients were diagnosed to have been infected with Helicobacter pylori (72.66%). Out of 89 patients with gastric ulcer, 61 patients were infected with Helicobacter pylori (68.53%). Forty two out of 51 patients (82.35%) with duodenal ulcers and 06 of 10 patients (60%) with carcinoma of stomach were positive for H. pylori. The remaining patients were found to be negative for the H. pylori infection.Conclusions: The findings of the study conclude that H. pylori was consistently associated with PUD.


2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Rania M. Kishk ◽  
Nashaat M. Soliman ◽  
Maha M. Anani ◽  
Nader Nemr ◽  
Ayman Salem ◽  
...  

Helicobacter pylori (H. pylori) plays a crucial role in the pathogenesis of gastritis, peptic ulcer, and gastric cancer. The presence of pathogenicity islands (PAI) genes contributes to the pathogenesis of many gastrointestinal disorders. Cytotoxin-associated gene A (cagA) and vacuolating cytotoxin gene (vacA) are the most known virulence genes in H. pylori. So, our aim was to study H. pylori virulence genes’ role in gastric disorders pathogenesis. Our study included 150 adult patients who suffered dyspeptic symptoms and were referred to the GIT endoscopy unit. Gastric biopsies were attained for rapid urease test (RUT) and histopathological examination, and multiplex PCR technique for detection of virulence genes was performed. It was found that 100 specimens were (RUT) positive, of which sixty samples (60%) were PCR positive for H. pylori ureC gene. The vacA and cagA genes were identified in 61.6% and 53% of H. pylori strains, respectively. Only 5 cases were vacA-positive and cagA-negative. The most virulent vacA s1 allele existed in 56.6% of cases. Out of the 60 H. pylori strains, 66% had at least one virulence gene and 34% did not show any virulence gene. H. pylori infection showed significant increase with age. H. pylori are prevalent amid dyspeptic patients in our region. The main genotype combinations were vacA+/cagA+ of s1m1 genotype and they were frequently associated with peptic ulcer diseases, gastritis, and gastroesophageal reflux disease.


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