Cell Culture-Taqman PCR Assay for Evaluation of Cryptosporidium ...

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Jul 19, 2002 - Alexandra R. Keegan,1* Stella Fanok,1 Paul T. Monis,1,2 and Christopher P. Saint2. The Co-operative ...... vine genotype. Emerg. Infect. Dis.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, May 2003, p. 2505–2511 0099-2240/03/$08.00⫹0 DOI: 10.1128/AEM.69.5.2505–2511.2003 Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Vol. 69, No. 5

Cell Culture-Taqman PCR Assay for Evaluation of Cryptosporidium parvum Disinfection Alexandra R. Keegan,1* Stella Fanok,1 Paul T. Monis,1,2 and Christopher P. Saint2 The Co-operative Research Centre for Water Quality and Treatment, Australian Water Quality Centre, South Australian Water Corporation, Salisbury, South Australia 5108,1 and School of Pharmaceutical, Molecular, and Biomedical Sciences, University of South Australia, Mawson Lakes, South Australia 5095,2 Australia 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