Molecular Characterization of Blastocystis Isolates From Human by Restriction Fragment Length Polymorphism (RFLP) Analysis

Study Overview
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Brief Summary
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Blastocystis is an anaerobic unicellular protozoal parasite infecting the gastrointestinal tract of humans and a wide range of animals. It is one of the most common enteric microorganisms with higher prevalence rates in developing than in developed countries. Feco-oral is the main route of transmission where low socioeconomic conditions, poor hygienic practices, close contact with animals, and drinking contaminated water act as major risk factors. Infection with Blastocystis was demonstrated in both symptomatic and asymptomatic people. For a long period, Blastocystis was considered a commensal organism with no pathogenic role, but recently, many studies linked it to different gastrointestinal symptoms such as nausea, diarrhea, and abdominal pain. Association with irritable bowel syndrome and colorectal cancer was also reported. This study aims to: - Identify subtypes of human Blastocystis isolates in Sohag by using RFLP-PCR and provide additional information on the molecular epidemiology of this parasite in our locality.

Official Title
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Molecular Characterization of Blastocystis Isolates From Human by Restriction Fragment Length Polymorphism (RFLP) Analysis in Sohag Governorate

Conditions
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Blastocystis Infections

Intervention / Treatment
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* Diagnostic Test: RFLP analysis of SSU rRNA


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Patients from outpatient clinics, Sohag hospitals. Exclusion Criteria: Patients received antiparasitic drugs in the last two weeks
Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Intervention/Treatment |
| --- |
|Diagnostic Test: RFLP analysis of SSU rRNA|Genomic DNA will be extracted from Blastocystis-positive fecal samples using a specific DNA Stool Mini Kit according to the manufacturer's instructions and will be stored at -20°C until use. PCR will be used to amplify a region of Blastocystis small subunit ribosomal RNA (SSU rRNA) gene. The amplified products for the SSU rRNA gene will be digested to determine the strain subtype of Blastocystis spp. using specific restriction enzymes.|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The number of cases with subtypes of human Blastocystis isolates in Sohag | by using RFLP-PCR | 14 weeks following the startpoint of the study. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The number of cases with Blastocystis infection is recorded. | By molecular examination of stool samples from 100 patients | 14 weeks following the startpoint of the study. | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Blastocystis, colorectal cancer, irritable bowel syndrome