Detection of Toxoplasma gondii oocysts in water sample concentrates by real-time PCR

Wenli Yang, H. D Alan Lindquist, Vitaliano Cama, Frank W. Schaefer, Eric Villegas, Ronald Fayer, Earl J. Lewis, Yaoyu Feng, Lihua Xiao

Research output: Contribution to journalArticlepeer-review

Abstract

PCR techniques in combination with conventional parasite concentration procedures have potential for the sensitive and specific detection of Toxoplasma gondii oocysts in water. Three real-time PCR assays based on the B1 gene and a 529-bp repetitive element were analyzed for the detection of T. gondii tachyzoites and oocysts. Lower sensitivity and specificity were obtained with the B1 gene-based PCR than with the 529-bp repeat-based PCR. New procedures for the real-time PCR detection of T. gondii oocysts in concentrates of surface water were developed and tested in conjunction with a method for the direct extraction of inhibitor-free DNA from water. This technique detected as few as one oocyst seeded to 0.5 ml of packed pellets from water samples concentrated by Envirocheck filters. Thus, this real-time PCR may provide a detection method alternative to the traditional mouse assay and microscopy.

Original languageEnglish (US)
Pages (from-to)3477-3483
Number of pages7
JournalApplied and Environmental Microbiology
Volume75
Issue number11
DOIs
StatePublished - Jun 2009
Externally publishedYes

ASJC Scopus subject areas

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

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