Citation:

Yang, W., H. LINDQUIST, V. Cama, F. W. SCHAEFER, E. VILLEGAS, R. Fayer, E. J. Lewis, Y. Feng, AND L. Xiao. Detection of Toxoplasma gondii oocuysts in water sample concentrates by real-time PCR. Journal of Environmental Microbiology. Blackwell Publishing, Malden, MA, 75(11):0099-2240, (2009).

Impact/Purpose:

PCR techniques in combination with conventional parasite concentration procedures have potential for 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 compared for detection of T. gondii tachyzoites and oocysts. Lower sensitivity and specificity were obtained using the B1 gene-based PCR than by the 529-bp repeat-based PCR. New procedures for real-time PCR detection of T. gondii oocysts in concentrates of surface water were developed and tested in conjunction with a method for 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.

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Journal Article

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