Development of a National Reference Facility for Cryptosporidium.pptVIP

Towards the Development of a National Reference Facility for CryptosporidiumCarolyn Read, Jenny Pender, Tom Murphy, John Egan, Grace Mulcahy and Theo de Waal. * Establishment of a Cryptosporidium reference facility located at Central Veterinary Research Laboratory, DAFF, Celbridge, Co Kildare Survey of Cryptosporidium monitoring in Irish drinking water supplies Detection of Cryptosporidium In Water- US EPA Method 1622 Genotyping of Cryptosporidium oocysts from slides using emerging methods Method for Detection and Genotyping Project Objectives Filtration IMS FITC Slide scraping DNA extraction Real-time PCR Traditional PCR High Resolution melt curve analysis Sequencing Progress to date Survey of Cryptosporidium monitoring in Irish drinking water supplies complete and available on request. USEPA method 1622 for the detection of Cryptosporidium oocysts in water has been established in the laboratory and validation is near completion. Routine genotyping of Cryptosporidium oocysts off slides from method 1622 by PCR-Sequencing has been used successfully for the pilot scheme samples Xiao et al., Ryan et al. Emerging methods using real time PCR followed by high resolution melt curve HRM analysis DiGiovanni et al. are being established. Pilot scheme sampling and analysis commenced in February 2011- 9 months completed. Preliminary Pilot Scheme Results DIC DAPI Cryptosporidium oocysts Future of Laboratory: Accreditation assessment by INAB due in March 2012 Commence genotyping on oocysts from slides referred from water testing laboratories Consultation with stakeholders to a establish model for future sustainability of a national reference facility for Cryptosporidium A Collaboration between the EPA, UCD and DAFF DiGiovanni et al, Water Research Foundation, Web Report #4099. Xiao et al, 2001, Applied and Environmental Microbiology, Mar 2001, p. 1097-1101 Ryan et al, 2003. Applied and Environmental Microbiolgoy, July 2003, P. 4302-4307 References