Antibiotyping and Prevalence of Virulent Genotypes Among Helicobacter Pylori and Their Impact on Response to Therapy
Trial Snapshot
- Phase
- Not Applicable
- Status
- Completed
- Sponsor
- Assiut University
- Enrollment
- 100
- Locations
- 1
- Primary Endpoint
- Relation of histopathological examination (gastritis classification) to clinical presentation and to resistance.
Study Overview
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.
Detailed Description
Helicobacter pylori (H. pylori) infection is one of the most common infections in humans, affecting more than half of the world population. New infections are thought to occur as a consequence of direct human-to-human transmission or environmental contamination.
The prevalence of the infection varies widely in rural developing areas (more than 80%) compared to urban developed ones (less than 40%), as a consequence of different socioeconomic and hygienic conditions .
The vertical mode is infection spread from ascendant to descendent within the same family, while horizontal transmission involves contact with individuals outside the family or environmental contamination.
Most individuals are infected by Helicobacter pylori during early childhood; in developing countries. Successful eradication is important to prevent the development of antibiotic resistance, as well as to reduce the number of treatments and procedures. Thus, national/ regional antibiotic resistance data could be used to guide treatment regimens for H pylori infection .
Several clinical factors associated with increased rates of antibiotic resistant H. pylori, including history of previous antibiotic exposure, increasing age, female gender, ethnicity/race, extent of alcohol use, and non-ulcer dyspepsia .
Study Design
- Study Type
- Interventional
- Allocation
- Na
- Intervention Model
- Single Group
- Primary Purpose
- Treatment
- Masking
- None
Eligibility Criteria
- Ages
- 18 Years to — (Adult, Older Adult)
- Sex
- All
- Accepts Healthy Volunteers
- No
Inclusion Criteria
- •- Any patient is above 18 years old with symptomatic H pylori infection patients diagnosed by positive H pylori Ag in stool (naïve treatment).
Exclusion Criteria
- •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.
Arms & Interventions
One group of naive H.Pylori infection will submitted for upper endoscopy.
Upper endoscopy will be done under complete septic condition and multiple gastric biopsies from corpus and antrum will be taken for :
- Histopathological examination.
- Culture and sensitivity of endoscopic biopsies.
- detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR).
- Then start empirical antibiotics regimens.
Intervention: Upper endoscopy (Procedure)
One group of naive H.Pylori infection will submitted for upper endoscopy.
Upper endoscopy will be done under complete septic condition and multiple gastric biopsies from corpus and antrum will be taken for :
- Histopathological examination.
- Culture and sensitivity of endoscopic biopsies.
- detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR).
- Then start empirical antibiotics regimens.
Intervention: empirical H.pylori regimens (Drug)
One group of naive H.Pylori infection will submitted for upper endoscopy.
Upper endoscopy will be done under complete septic condition and multiple gastric biopsies from corpus and antrum will be taken for :
- Histopathological examination.
- Culture and sensitivity of endoscopic biopsies.
- detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR).
- Then start empirical antibiotics regimens.
Intervention: H.Pylori Ag in stool (Diagnostic Test)
One group of naive H.Pylori infection will submitted for upper endoscopy.
Upper endoscopy will be done under complete septic condition and multiple gastric biopsies from corpus and antrum will be taken for :
- Histopathological examination.
- Culture and sensitivity of endoscopic biopsies.
- detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR).
- Then start empirical antibiotics regimens.
Intervention: 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)
One group of naive H.Pylori infection will submitted for upper endoscopy.
Upper endoscopy will be done under complete septic condition and multiple gastric biopsies from corpus and antrum will be taken for :
- Histopathological examination.
- Culture and sensitivity of endoscopic biopsies.
- detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR).
- Then start empirical antibiotics regimens.
Intervention: Culture and sensitivity of endoscopic gastric biopsies: (Diagnostic Test)
One group of naive H.Pylori infection will submitted for upper endoscopy.
Upper endoscopy will be done under complete septic condition and multiple gastric biopsies from corpus and antrum will be taken for :
- Histopathological examination.
- Culture and sensitivity of endoscopic biopsies.
- detection of vacuolating cytotoxin A (Vac A) and cytotoxin-associated gene A (Cag A) virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR).
- Then start empirical antibiotics regimens.
Intervention: Histopathological examination endoscopic gastric biopsies: (Other)
Outcomes
Primary Outcomes
Relation of histopathological examination (gastritis classification) to clinical presentation and to resistance.
Time Frame: up to 12 months
Histopathological examination of endoscopic biopsies and detect acute or chronic inflammation induced by H.pylori bacteria.
detection of pre treatment antibiotic culture and sensitivity test
Time Frame: up to 12 months
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.
detection of vacuolating cytotoxin A and cytotoxin-associated gene A virulent Helicobacter Pylori genotypes by polymerase chain reaction amplification(PCR)
Time Frame: up to 12 months
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.
Secondary Outcomes
- Detection of diet and drugs effect on response to treatment(up to 12 months)
Investigators
Nariman Zaghloul Bekhiet
Doctor
Assiut University