Evaluation of an alternative IMS dissociation procedure for use with Method 1622: detection of <i>Cryptosporidium</i> in water

نویسندگان

  • Michael W. Ware
  • Larry Wymer
  • Alan Lindquist
  • Frank W. Schaefer
  • H. D. Alan Lindquist
چکیده

U.S.EPA Methods 1622 and 1623 are used to detect and quantify Cryptosporidium oocysts in water. 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 4V,6-diamidino-2-phenyl indole dihydrochloride (DAPI). The purpose of this study was to evaluate a new IMS dissociation, a 10-min incubation at 80 jC. Heat dissociation improved the average oocyst recovery from 41% to 71% in seeded reagent water, and from 10% to 51% in seeded river samples. The average DAPI confirmation rate improved from 49% to 93% in reagent water, and from 48% to 73% in river samples. This modification improved both oocyst recovery and confirmation. Published by Elsevier B.V.

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تاریخ انتشار 2017