Antibiotyping and Prevalence of Virulent Genotypes Among Helicobacter Pylori and Their Impact on Response to Therapy
- Conditions
- H Pylori Infection
- Interventions
- Procedure: Upper endoscopyDrug: empirical H.pylori regimensDiagnostic Test: H.Pylori Ag in stool
- Registration Number
- NCT05444439
- Lead Sponsor
- Assiut University
- 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 .
The primary antibiotic resistance of H. pylori is increasing worldwide. The overall resistance rate was found to be 4.55% for amoxicillin; 27.22% for clarithromycin; 39.66% for metronidazole; and 22.48% for levofloxacin.
Therefore, drug susceptibility result of these antibiotics is necessary to select the appropriate drug for the successful eradication of the infection.
Helicobacter pylori exhibit specific geographic distributions that are related to clinical outcomes. Despite the high infection rate of H. pylori throughout the world, the genetic epidemiology surveillance of H. pylori still needs to be improved.
Recruitment & Eligibility
- Status
- UNKNOWN
- Sex
- All
- Target Recruitment
- 100
- Any patient is above 18 years old with symptomatic H pylori infection patients diagnosed by positive H pylori Ag in stool (naïve treatment).
- 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.
Study & Design
- Study Type
- INTERVENTIONAL
- Study Design
- SINGLE_GROUP
- Arm && Interventions
Group Intervention Description One group of naive H.Pylori infection will submitted for upper endoscopy. Upper endoscopy 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. One group of naive H.Pylori infection will submitted for upper endoscopy. empirical H.pylori regimens 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. One group of naive H.Pylori infection will submitted for upper endoscopy. H.Pylori Ag in stool 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.
- Primary Outcome Measures
Name Time Method Relation of histopathological examination (gastritis classification) to clinical presentation and to resistance. 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 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) 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 Outcome Measures
Name Time Method Detection of diet and drugs effect on response to treatment up to 12 months Detect diet intake (fatty and spicy meal), drugs use (as NSAIDs, antibiotics, proton pump inhibitors in last month) by Questionnaire.
Trial Locations
- Locations (1)
Nariman Zaghloul Bekhiet
🇪🇬Assiut, Egypt