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

J Microbiol Methods. 2003 Dec;55(3):575-83. doi: 10.1016/j.mimet.2003.06.001.

Abstract

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 4',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 degrees C. 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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cryptosporidium parvum / isolation & purification*
  • Environmental Monitoring / methods*
  • Flow Cytometry
  • Fluorescent Antibody Technique / methods
  • Fluorescent Dyes / metabolism
  • Hydrogen-Ion Concentration
  • Immunomagnetic Separation / methods*
  • Indoles / metabolism
  • Multivariate Analysis
  • Oocysts / isolation & purification
  • United States
  • United States Environmental Protection Agency
  • Water / parasitology*

Substances

  • Fluorescent Dyes
  • Indoles
  • Water
  • DAPI