Detection and enumeration of Cryptosporidium oocysts in environmental water samples by Real-time PCR assay

detection and enumeration of cryptosporidium oocysts in environmental water samples by real-time pcr assay

introduction: the protozoan parasite, cryptosporidium spp., widely spreads in both raw and drinking waters. it is the causative agents of waterborne diarrhea and gastroenteritis in the world. in the present study, a molecular assay was used for the detection and quantification of cryptosporidium oocysts in environmental water samples. materials and methods: thirty surface water samples were col...

Sensitive detection of Cryptosporidium oocysts in environmental water samples by reverse transcription-PCR.

Pupply. With a reverse transcription-PCR (RT-PCR) amplifylng heat shock protein 70 (hsp70) gene, we were able to detect two differenttypes of viable Cyyptosporidium oocysts from the environment. Water specimens were filtered under positive pressure through a 3-Elm polycarbonate filter (Nuclepore filter, Whatman, Maidstone, England). The trapped materials were detached from the filter by sonicat...

Evaluation of TaqMan real-time PCR for the detection of viable Cryptosporidium parvum oocysts in environmental water samples

a nested-pcr assay for detection of cryptosporidium parvum oocysts in water samples

cryptosporidiosis is a gastroenteric disease caused by the protozoan parasite cryptosporidium parvum. water–borne transmission of this organism has become more prevalent in recent years. current method for detection of c. parvum oocysts in water is immunofluoresence assay (ifa). the method is time consuming, laborious and particularly not–specific. it cannot determine the infectivity of detecte...

Detection and genotyping of oocysts of Cryptosporidium parvum by real-time PCR and melting curve analysis.

Several real-time PCR procedures for the detection and genotyping of oocysts of Cryptosporidium parvum were evaluated. A 40-cycle amplification of a 157-bp fragment from the C. parvum beta-tubulin gene detected individual oocysts which were introduced into the reaction mixture by micromanipulation. SYBR Green I melting curve analysis was used to confirm the specificity of the method when DNA ex...

Recovery of Cryptosporidium from spiked water and stool samples measured by PCR and real time PCR

Cryptosporidium parvum is a common intestinal protozoan parasite infecting humans and a wide range of animals, whose diagnostics present considerable difficulties. These arise from the exceptionally robust nature of the oocyst’s walls, which necessitates more stringent treatments for disruption and recovery of DNA for analysis using molecular methods. In the case of water, which is the major so...