Isolation of Small Number of Cryptosporidium parvum Oocyst Using Immunochromatography. 2014

Elahe Ebrahimzade, and Parviz Shayan, and Zeinab Asghari, and Sedighe Jafari, and Zahra Omidian
Department of Parasitology, Iranian Center for Ticks and Tick-borne Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.

BACKGROUND Cryptosporidium parvum causes severe gastroenteritis in immunocompromised human and new borne animals. The organism can be transmitted through water. Since small number of C. parvum is infectious, the aim of the present study was to develop a chromatography method for the isolation of C. parvum oocyst in samples with limited number of oocysts. METHODS Antibody was prepared against whole antigen from C. parvum oocysts, the achieved Ab bound to the sepharose 4B and used for the isolation of oocysts. Antibody against P23 bound to the sepharose 4B, used also for the isolation of C. parvum oocyst. In comparison to these both methods, 2 traditional methods (Salt floatation and 55% sucrose floatation) were also performed. RESULTS Both chromatography methods could bind oocysts with capacity depends on the column size. The isolated oocysts were free of bacteria. Our results showed that the traditional methods are useful for the isolation of oocysts from feces, in its smear stained with ziehl-nelsen, at least 3 oocyts are detectable in each microscopic field under 1000 X magnification. In contrast to the chromatography methods, the bacterial contamination was always observed in oocysts isolated with traditional methods. CONCLUSIONS Immunochromatography could be used for the successful isolation of C. parvum oocysts from the samples containing limited number of oocysts.

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