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


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.

SUBMITTER: Yang W 

PROVIDER: S-EPMC2687264 | biostudies-literature | 2009 Jun

REPOSITORIES: biostudies-literature

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

Yang Wenli W   Lindquist H D Alan HD   Cama Vitaliano V   Schaefer Frank W FW   Villegas Eric E   Fayer Ronald R   Lewis Earl J EJ   Feng Yaoyu Y   Xiao Lihua L  

Applied and environmental microbiology 20090410 11


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  ...[more]

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