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Enhanced Dapi Staining for Cryptosporidium in Water Samples

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Abstract:The U.S. Environmental Protection Agency's Method 1623 is used to detect and quantify the presence of {ital Cryptosporidium} spp. oocysts in water. The protocol consists of concentrating a sample, staining this concentrate with a fluorescent antibody, and examining the sample microscopically. Since algae can be misinterpreted as positives using this procedure, confirmation may be done by either differential interference contrast microscopy, or by observation of the staining pattern when stained with 4,6-diamidino 2-phenyl-indole dihydrochloride (DAPI). Our research has shown that differential interference contrast is only able to confirm about 2% of oocysts with sporozoites in a fresh preparation. The rate of confirmation is improved to around 30% with DAPI staining when following the Method 1623 protocol. By a slight modification of the DAPI protocol, the confirmation rate improves to nearly 100%. A number of formats for optimizing slide preparation have been tested. Confirmation by both fluorescent antibody and DAPI staining is also possible using flow cytometry.
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Citation:Ware, M. W., F. W. Schaefer III, and H. A. Lindquist. Enhanced Dapi Staining for Cryptosporidium in Water Samples. Presented at American Society for Parasitologists Annual Meeting, Albuquerque, NM, June 29-July 3, 2001.
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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: Abstrct/Oral
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Presented: 06/29/2001
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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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