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PP 6.25
Real- Time PCR and Flow Cytometry for Detection of Cyclospora Oocysts from
Fecal Samples of Gastrointestinal Symptomatic and Asymptomatic Patients
Eman M. Hussein¹, Amal A. El-Moamly¹, Hamdy A. Dawoud¹ and Hanaa Fahmy ²
Departments of Parasitology1 and Clinical Pathology², Faculty of Medicine, Suez Canal
University, smailia, Egypt.
Objectives
Cyclospora cayetanensis has emerged as an important human pathogen that causes
enteric disease in both immunocompromized and immunocompotent hosts. Also,
asymptomatic patients excrete oocysts in their stool were identified. Until Now, the
difference in parasitic load among cyclosporaisis patients was not detected. In this study,
the magnitude of oocysts excretion in relation to the pathological severity was assessed
among cyclosporiasis patients with using Flow Cytometry and quantitative Real- time
PCR.
Methods
Oocysts from stool samples of 25 symptomatic and 10 asymptomatic gastrointestinal
cyclosporiasis patients were identified by both modified Acid Fast Trichrome and modified
Ziehl-Neelsen staining methods and were confirmed by its auto-fluorescent
characterizations. In addition, 10 persons with negative stool samples were selected as
control group. The intensity of infection was calculated by number of oocysts/200
microscopic filed with immersion 400. Flow cytometry and quantitative Real- time PCR
were used to assess the relation between the degree of infection and magnitude of
oocysts excretions. Statistical analysis was applied using X2.
Results
The degrees of infection among symptomatic patients were identified by staining methods
as mild in 16%, moderate in 24% and severe in 60%. All of asymptomatic patients were
in mild infection degree. Flow cytometry was conducted to all samples and Cyclospora
oocysts were identified in 100% and 80% of symptomatic and asymptomatic patients
respectively. This difference were statistically significant (P