Detection of Giardia cysts with a cDNA probe and applications to water samples

M. Abbaszadegan, C. P. Gerba, J. B. Rose

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

Giardiasis is the most common human parasite infection in the United States, causing a lengthy course of diarrhea. Transmission of Giardia species is by the fecal-oral route, and numerous waterborne outbreaks have been documented. The Environmental Protection Agency has regulated Giardia species in drinking water through the Surface Water Treatment Rule. Current methods for detection of Giardia species in water rely primarily on microscopic observation of water concentrates with immunofluorescence techniques. We evaluated the efficacy of using a gene-specific probe for the detection of Giardia species in water. A cDNA probe, 265 bp long, from the small subunit of rRNA of Giardia lamblia was used for detection of cysts. The replicative form of the M13 vector with an insert was isolated from lysed host Escherichia coli XL1-Blue and used for production of the cDNA probe by nick translation with 32P-labeled nucleotides. Six different protocols were tested for extracting nucleic acids from the cysts. With the most efficient procedure, disrupting Giardia cysts with glass beads in the presence of proteinase K, as few as 1 to 5 cysts per ml can be detected in water sample concentrates with dot blot hybridization assays.

Original languageEnglish (US)
Pages (from-to)927-931
Number of pages5
JournalApplied and environmental microbiology
Volume57
Issue number4
DOIs
StatePublished - 1991
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Fingerprint Dive into the research topics of 'Detection of Giardia cysts with a cDNA probe and applications to water samples'. Together they form a unique fingerprint.

Cite this