Antibiotyping and Prevalence of Virulent Genotypes Among Helicobacter Pylori and Their Impact on Response to Therapy

NCT05444439 | Completed DMC

• 1 other identifier: APVGHP
• Study Type: interventional
• Target: 100
• Locations: 1 country
• Sites: 1

Brief Summary

• Detection of primary antimicrobial susceptibility and resistance of Helicobacter Pylori infection.
• Detection of resistance and virulence genes of Helicobacter Pylori infection.
• Assessment of H pylori carcinogenicity gene.
• Evaluation of outcome and efficacy of antibiotics regimen will be used in our research.
• Evaluation of effect of other factors as diet (fatty and spicy meal), drugs as NSAIDs use, antibiotics for any cause on response of H pylori to antibiotics regimen.

Conditions

H Pylori Infection

Keywords

H Pylori Infection

Outcome Measures

Primary Outcomes (3)

• detection of pre treatment antibiotic culture and sensitivity test

  During endoscopy, three biopsies will be taken from antrum and/or corpus of stomach, Biopsies will be examined for identification of H. pylori by rapid urease test also known as the Campylobacter-like organism test (Kimberly-Clark, USA). Culture of the bacterium on Columbia agar (Oxoid-UK) plus 5-7% defibrinated horse or sheep blood and selective Dent supplement (Oxoid-UK) under microaerophilic condition at 37 degree Celsius for 3-5days. So the investigators will detect prevalence and types of antibiotic H.Pylori resistance in biopsies.

  up to 12 months

• detection of vacuolating cytotoxin A and cytotoxin-associated gene A virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR)

  Extraction of DNA, using Wizard® Genomic DNA Purification Kit (Promega-USA), will be done following the manufacturer instructions. For detection of vacuolating cytotoxin A and cytotoxin-associated gene A gene, polymerase chain reaction amplification will be performed with a 9 minutes' initial denaturation at 94˚C, followed by 35 cycles of 1 minute at 94˚C, 45 seconds at 60˚C, and 45 seconds at 72˚C. Final extension will be performed for 5 minutes at 72˚C. For detection of vacuolating cytotoxin A (S1/S2, m1, m2), PCR will be performed with a 9 minutes' initial denaturation at 94˚C, followed by 35 cycles of 1 minute at 94˚C, 45 seconds at 56˚C, and 45 seconds at 72˚C. Final extension will be performed for 5 minutes at 72˚C. Amplified DNA will be analyzed by agarose gel electrophoresis. A positive sample will give bands at DNA fragment 138-bp for cytotoxin-associated gene A, at 259/286-bp for vacuolating cytotoxin A s1/s2, and 290-bp and 352-bp for m1 \& m2, respectively.

  up to 12 months

• Relation of histopathological examination (gastritis classification) to clinical presentation and to resistance.

  Histopathological examination of endoscopic biopsies and detect acute or chronic inflammation induced by H.pylori bacteria.

  up to 12 months

Secondary Outcomes (1)

• Detection of diet and drugs effect on response to treatment

  up to 12 months

Study Arms (1)

One group of naive H.Pylori infection will submitted for upper endoscopy.

OTHER

Upper endoscopy will be done under complete septic condition and multiple gastric biopsies from corpus and antrum will be taken for : 1. Histopathological examination. 2. Culture and sensitivity of endoscopic biopsies. 3. detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR). 4. Then start empirical antibiotics regimens.

Genetic: detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent H.Pylori genotypes by polymerase chain reaction amplification(PCR) in gastric biopsies :; Diagnostic Test: Culture and sensitivity of endoscopic gastric biopsies:; Other: Histopathological examination endoscopic gastric biopsies:; Procedure: Upper endoscopy; Drug: empirical H.pylori regimens; Diagnostic Test: H.Pylori Ag in stool

Interventions

Culture and sensitivity of endoscopic gastric biopsies: DIAGNOSTIC_TEST

During endoscopy, three biopsies will be taken from antrum and/or corpus of and will be examined by rapid urease test also known as Campylobacter-like organism test (Kimberly-Clark,USA). Culture of the bacterium on Columbia agar plus 5-7% defibrinated horse or sheep blood and selective Dent supplement under microaerophilic condition at 37degree Celsius for 3-5days.

One group of naive H.Pylori infection will submitted for upper endoscopy.

Histopathological examination endoscopic gastric biopsies: OTHER

Histological sections from the antrum and body regions will be stained with Giemsa stain. Giemsa stain working solution was prepared as follows: 40 ml Giemsa stock solution with 60 ml of distilled water. Giemsa stock solution will be prepared as follows: Giemsa powder 4 g, glycerol 250 ml and methanol 250 ml. Histological sections will be examined by a gastrointestinal pathologist to standardize the classification of gastritis. Histopathological classification will be recorded as following (acute gastritis, mild-moderate- severe chronic gastritis, Glandular atrophy or Intestinal metaplasia).

One group of naive H.Pylori infection will submitted for upper endoscopy.

Upper endoscopy PROCEDURE

it will be done patient before start empirical therapy and multiple antrum and/or corpus biopsies will be taken . Patient will be fasting at least 8 hours.Upper and findings will be recorded in the report as presence of gastroesophageal reflux disease, diffuse or localised gastric and/or duodenal redness, mucosal swelling, atrophy of mucosa, nodularity, intestinal metaplasia, erosions or ulcers.

One group of naive H.Pylori infection will submitted for upper endoscopy.

empirical H.pylori regimens DRUG

Start empirical antibiotics regimens patient triple therapy ((levofloxacin 400 mg once , amoxicillin 1000 mg twice for 2 weeks) and (Proton pump inhibitor twice for 1 month)). Follow up after 2 weeks after finishing regimen by Stool Ag in stool

One group of naive H.Pylori infection will submitted for upper endoscopy.

H.Pylori Ag in stool DIAGNOSTIC_TEST

Any symptomatic patient is above 18 years old with H pylori infection patients diagnosed by positive H pylori Ag in stool will be included. assessment of laboratory response, 2 weeks after finishing empirical therapy.

One group of naive H.Pylori infection will submitted for upper endoscopy.

detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent H.Pylori genotypes by polymerase chain reaction amplification(PCR) in gastric biopsies : GENETIC

Extraction of DNA, using Wizard® Genomic DNA Purification Kit (Promega-USA), will be done following the manufacturer instructions. For detection of vacuolating cytotoxin A and cytotoxin-associated gene A gene, polymerase chain reaction amplification will be performed with a 9 minutes' initial denaturation at 94˚C, followed by 35 cycles of 1 minute at 94˚C, 45 seconds at 60˚C, and 45 seconds at 72˚C. Final extension will be performed for 5 minutes at 72˚C. For detection of vacuolating cytotoxin A (S1/S2, m1, m2), PCR will be performed with a 9 minutes' initial denaturation at 94˚C, followed by 35 cycles of 1 minute at 94˚C, 45 seconds at 56˚C, and 45 seconds at 72˚C. Final extension will be performed for 5 minutes at 72˚C. Amplified DNA will be analyzed by agarose gel electrophoresis. A positive sample will give bands at DNA fragment 138-bp for cytotoxin-associated gene A, at 259/286-bp for vacuolating cytotoxin A s1/s2, and 290-bp and 352-bp for m1 \& m2, respectively

One group of naive H.Pylori infection will submitted for upper endoscopy.

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: No
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• \- Any patient is above 18 years old with symptomatic H pylori infection patients diagnosed by positive H pylori Ag in stool (naïve treatment).

You may not qualify if:

• Patients who aren't eligible to endoscopy.
• Patients are under 18 years old
• Patient's refusal
• history of antibiotics or proton pump inhibitor use during last month.

Sponsors & Collaborators

• Assiut University: lead

Study Sites (1)

Nariman Zaghloul Bekhiet

Asyut, Egypt

Location

Related Publications (3)

• Liu Y, Wang S, Yang F, Chi W, Ding L, Liu T, Zhu F, Ji D, Zhou J, Fang Y, Zhang J, Xiang P, Zhang Y, Zhao H. Antimicrobial resistance patterns and genetic elements associated with the antibiotic resistance of Helicobacter pylori strains from Shanghai. Gut Pathog. 2022 Mar 30;14(1):14. doi: 10.1186/s13099-022-00488-y.

  BACKGROUND

• Deng L, He XY, Tang B, Xiang Y, Yue JJ. An improved quantitative real-time polymerase chain reaction technology for Helicobacter pylori detection in stomach tissue and its application value in clinical precision testing. BMC Biotechnol. 2020 Jun 22;20(1):33. doi: 10.1186/s12896-020-00624-z.

  BACKGROUND

• Pokhrel N, Khanal B, Rai K, Subedi M, Bhattarai NR. Application of PCR and Microscopy to Detect Helicobacter pylori in Gastric Biopsy Specimen among Acid Peptic Disorders at Tertiary Care Centre in Eastern Nepal. Can J Infect Dis Med Microbiol. 2019 Feb 5;2019:3695307. doi: 10.1155/2019/3695307. eCollection 2019.

  BACKGROUND

Related Links

• Related Info
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MeSH Terms

Interventions

Gastroscopy; Defecation

Intervention Hierarchy (Ancestors)

Endoscopy, Gastrointestinal; Endoscopy, Digestive System; Diagnostic Techniques, Digestive System; Diagnostic Techniques and Procedures; Diagnosis; Endoscopy; Diagnostic Techniques, Surgical; Digestive System Surgical Procedures; Surgical Procedures, Operative; Minimally Invasive Surgical Procedures; Digestive System Physiological Phenomena; Digestive System and Oral Physiological Phenomena

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NA
• Masking: NONE
• Purpose: TREATMENT
• Intervention Model: SINGLE GROUP
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Doctor

Model Details: Histopathological examination Of gastric endoscopic biopsies Detection of cag A and vac A Genotypes by PCR in endoscopic biopsies Culture and sensitivity of endoscopic biopsies. Start empirical antibiotics regimens patient triple therapy ((levofloxacin 400 mg once , amoxicillin 1000 mg twice for 2 weeks) and (Proton pump inhibitor twice for 1 month)) then follow up by H. pylori Ag in stool.

Study Record Dates

• First Submitted: June 14, 2022
• First Posted: July 6, 2022
• Study Start: January 20, 2024
• Primary Completion: September 15, 2024
• Study Completion: January 20, 2025
• Last Updated: March 13, 2026

Record last verified: 2022-06

Locations

EG (1)