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Applied and Environmental Microbiology, May 2003, p. 2505-2511, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2505-2511.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Cell Culture-Taqman PCR Assay for Evaluation of Cryptosporidium parvum Disinfection

The Co-operative Research Centre for Water Quality and Treatment, Australian Water Quality Centre, South Australian Water Corporation, Salisbury, South Australia 5108,¹ School of Pharmaceutical, Molecular, and Biomedical Sciences, University of South Australia, Mawson Lakes, South Australia 5095, Australia²

Received 19 July 2002/ Accepted 28 January 2003

Cryptosporidium parvum represents a challenge to the water industry and a threat to public health. In this study, we developed a cell culture-quantitative PCR assay to evaluate the inactivation of C. parvum with disinfectants. The assay was validated by using a range of disinfectants in common use in the water industry, including low-pressure UV light (LP-UV), ozone, mixed oxidants (MIOX), and chlorine. The assay was demonstrated to be reliable and sensitive, with a lower detection limit of a single infectious oocyst. Effective oocyst inactivation was achieved (>2 log₁₀ units) with LP-UV (20 mJ/cm²) or 2 mg of ozone/liter (for 10 min). MIOX and chlorine treatments of oocysts resulted in minimal effective disinfection, with <0.1 log₁₀ unit being inactivated. These results demonstrate the inability of MIOX to inactivate Cryptosporidium. The assay is a valuable tool for the evaluation of disinfection systems for drinking water and recycled water.

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