Development of sensitive detection of cryptosporidum and giardia from surface water in Iran

ABSTRACT

The protozoan parasites Cryptosporidium spp. and Giardia are known to occur widely in both raw and drinking waters. They are two of the causative agents of waterborne outbreaks of gastroenteritis throughout the world. In the present study, a PCR assay and FA were developed for detection of Cryptosporidium oocysts and Giardia cyst in environmental samples. We have detected Cryptosporidium spp. oocysts and Giardia cysts in seeded and unseeded environmental water samples by PCR method. Water samples were spiked with oocysts [50, 100, 300, 500] and filtrated with a 1.2- micro m pore size cellulos nitrate and follow by DNA extraction and purification by QIA amp DNA mini kit. Nested -PCR assay amplified an 850 bp fragment of 18s rDNA gene specific for Cryptosporidium and 453 bp fragment of glutamate dehydrogenase [GDH] target genre for Giardia. Also many river water from north of Iran, be checked by these methods. Cryptosporidium and Giardia DNAs were detected in seeded water sample and Giardia was detected in all 5 water samples from river in north of Iran by nested-PCR and FA. Also in one river water sample, Cryptosporidium was detected. This protocol if effective for detection of these waterborne parasites in treated and untreated water samples. This study can also serve as a platform for further investigations and research water source in Iran