Auramine Phenol Staining Technique for Revealing Different Coccidian Parasites

Study Overview
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Brief Summary
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The increased number of documented human coccidian infections, including Cryptosporidium parvum, Cyclospora cayetanensis, Isospora belli, and Sarcocystis spp., that are often indistinguishable from other forms of community-acquired diarrhea, together with the possibility of treating some of them, suggests a need for proper diagnostic techniques to recover and identify these organisms

Detailed Description
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The increased number of documented human coccidian infections, including Cryptosporidium parvum, Cyclospora cayetanensis, Isospora belli, and Sarcocystis spp., that are often indistinguishable from other forms of community-acquired diarrhea, together with the possibility of treating some of them, suggests a need for proper diagnostic techniques to recover and identify these organisms. Earlier, Cryptosporidium and Cystoisospora were assumed to be the causative agents of acute diarrhea in animals but recently have emerged as one of the leading causes of prolonged lifethreatening diarrhea in immunocompromised patients particularly in those with immune dysfunction like AIDS who may show severe intestinal injury, prolonged diarrhea, extreme weight loss, and generalized wasting. In contrast, healthy individuals commonly present with mild to moderate self-limiting diarrhea during the infective stage, besides asymptomatic infection can also occur. Detection of coccidian parasites is mostly through microscopic observation using Kinyon's acid-fast stain. Although Sheather sugar flotation may result in increased concentration of the cysts, this method is cumbersome and does not lend itself to convenient incorporation within the routine concentration and staining procedures favored in most clinical laboratories. Any acid-fast stain will be taken up by cyst walls, but the time required to prepare and examine acid-fast stains on all stool samples received for routine parasitology would not be cost-effective unless the prevalence of coccidian parasites was shown to warrant such effort.

Official Title
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Auramine Phenol Staining Technique for Revealing Different Coccidian Parasites

Conditions
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Chronic Diarrhea

Intervention / Treatment
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* Diagnostic Test: auramine phenol staining


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: patients complaining of chronic diarrhea . Exclusion Criteria: patients taking antibiotics within the previous four weeks, and anti-parasitic drugs within the previous two weeks.

Ages Eligible for Study
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Minimum Age: 6 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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All

Study Plan
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How is the study designed?
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Arms and Interventions

| Intervention/Treatment |
| --- |
|Diagnostic Test: auramine phenol staining|The samples will be divided into 2 parts. The 1st part will be subjected to a direct wet saline smear, iodine smear, and concentration techniques. The 2nd part will be used to perform thin smears of the fecal concentrates and air-dried on 2 different slides. The first slide will be stained with Kinyoun's acid-fast stain and examined with the conventional light microscope and the second will be stained with auramine-phenol stain and examined with the fluorescent microscope.|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| prevalence of coccidian parasites | number of patients positive for coccidian parasites | 1 month | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| auramine phenol staining | accuracy of auramine phenol staining in the detection of coccidian parasites in comparison to light microscopy | 1 month | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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coccidian parasites, auramine phenol staining, fluorescence microscopy, modified Ziehl Nelseen, Cryptosporidium