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A QUANTITATIVE REAL-TIME PCR ASSAY FOR DETECTION OF CYCLOSPORA CAYETANESIS
Citation:
Varma, M, H.D A. Lindquist, F W. Schaefer III, M W. Ware, AND J. D. Hester. A QUANTITATIVE REAL-TIME PCR ASSAY FOR DETECTION OF CYCLOSPORA CAYETANESIS. Presented at VII International Workshops on Opportunistic Protists (IWOP), Cincinnati, OH, June 13-16, 2001.
Impact/Purpose:
The main objective is to develop a method to detect Cyclospora in water.
Description:
Cyclospora cayetanensis, a coccidian parasite with a direct fecal-oral life cycle, has become recognized world-wide as an emerging pathogen in both immunocompromised and immunocompetent individuals. Clinical manifestations include prolonged diarrhea, nausea, abdominal cramps, anorexia, weight loss and other symptoms of gastroenteritis. While most cases of infection with C. cayetanensis have been associated with food-borne transmission, contamination of water has also been observd. Until now, there has been no standardized protocol for the detection of C. cayetanensis oocysts in the environment.
We report for the first time the development and application of a real-time, quantitative PCR assay for the detection of C. cayetanensis in a variety of human clinical speciments as well as in water concentrates seeded with C. cayetanensis oocysts. The specificity of this oligonucleotide primer set and probe is derived from the genetic variability between the rDNA of C. cayetanensis and other organisms. The real-time PCR assay has been optimized specifically to detect a single C. cayetanensis oocyst. This molecular technique can be easily adapted to detect this parasite from a wide variety of food, clinical and environmental samples, as it is robust, reliable, and sensitive.