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Evaluation of An Alternative Ims Dissociation Procedure for Use With Method 1622: Detection of Cryptosporidium in Water

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Abstract:U.S. EPA Method 1623 is used to detect and quantify Cruptosporidum spp. oocysts in ater. The protocol consists of filtration, immunomagnetic separation (IMS), staining with a fluorescent antibody, and microscopic analysis. Microscopic analysis includes detection by fluorescent antibody and confirmation by the demonstration of 1-4 sporozoites or nuclei after staining with 4',6-diamidino-2-phenyl indole dihydrochloride (DAPI). Confirmation is required because some algae appear to be oocysts when using this fluorescent antibody method. The purpose fo this study was to develop and evaluate a new IMS dissociation using a 10 minute incubation at 80 degrees C. Using this adaptation, the average oocyst recovery rate improved from 41% to 71% in seeded reagent water samples, and from 10% to 51% in seeded river samples. The average oocyst confirmation rate by DAPI improved from 49% to 93% in reagent water samples and from 48% to 73% in river water samples. This modification improved Method 1623 by increasing both the oocyst recovery and DAPI confirmation rates.
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Citation:Ware, M. W., L. Wymer, H. A. Lindquist, and F. W. Schaefer III. Evaluation of An Alternative Ims Dissociation Procedure for Use With Method 1622: Detection of Cryptosporidium in Water. WATER RESEARCH. Elsevier Science Ltd, New York, NY, 55(3):575-583, (2003).
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Contact: Mary P. O'Bryant - (919)-541-4871 or obriant.mary@epa.gov
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Division: Microbiological & Chemical Exposure Assessment Division
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Branch: Biohazard Assessment Research Branch
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Product Type: Journal
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Published: 12/01/2003
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Related Entries:
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Bullet Item Detecting Ccl-Related, Emerging and Regulated Waterborne Human Protozoa for Exposure Assessment
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Last Updated on Monday, October 22, 2007
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