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In JoVE (1)
Other Publications (1)
Articles by Eric R. Rhodes in JoVE
JoVE Immunology and Infection
A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.
1National Exposure Research Laboratory, Office of Research and Development, US Environmental Protection Agency, 2Shaw Environmental & Infrastructure, 3Office of Ground Water and Drinking Water, US Environmental Protection Agency
This protocol describes the use of a tangential flow hollow-fiber ultrafiltration sample concentration system and a heat dissociation as alternative steps for the detection of waterborne Cryptosporidium and Giardia species using EPA Method 1623.
Other articles by Eric R. Rhodes on PubMed
Evaluation of Hollow-fiber Ultrafiltration Primary Concentration of Pathogens and Secondary Concentration of Viruses from Water
Tangential flow hollow-fiber ultrafiltration (HFUF) was evaluated for virus and Cryptosporidium parvum concentration from water. Recovery of viruses at a low filtration rate was found to be significantly greater than at a higher filtration rate, with the recoveries of bacteriophage MS2 at high and low filtration rates shown to be 64.7% and 98.7%, respectively. Poliovirus recoveries from tap water were similar to MS2, with recoveries of 62.9% and 104.5% for high and low filtration rates, respectively. C. parvum, which was only tested at high filtration rates, had an average recovery was 105.1%. In addition to the optimization of the primary concentration technique, this study also compared several secondary concentration procedures. The highest recovery (89.5%) of poliovirus from tap water concentrates was obtained when a beef extract-celite method was used and the virus was eluted from the celite with phosphate buffered saline, pH 9.0. When HFUF primary concentration and the optimal secondary concentration methods were combined, an average recovery of 97.0 ± 35.6% or 89.3 ± 19.3%, depending on spike level, was achieved for poliovirus. This study demonstrated that HFUF primary concentration method is effective at recovering MS2, poliovirus and C. parvum from large volumes of water and that beef extract-celite method is an effective secondary concentration method for the poliovirus tested.
