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Applied and Environmental Microbiology, November 2007, p. 7474-7476, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.01652-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of Two DNA Template Preparation Methods for Post-Immunomagnetic Separation Detection of Cryptosporidium parvum in Foods and Beverages by PCR{triangledown}

Christian D. Frazar and Palmer A. Orlandi*

Division of Virulence Assessment, Center for Food Safety and Applied Nutrition, Food and Drug Administration, Laurel, Maryland 20708

Received 19 July 2007/ Accepted 10 September 2007

Cryptosporidium parvum oocysts were recovered by immunomagnetic separation from six artificially contaminated foods. Two DNA isolation methods were subsequently evaluated by PCR. The FTA Concentrator-PS filter provided rapid and reproducible detection, although variability increased at lower inoculum levels (88% and 15% detection in high- and low-inoculum-level samples, respectively). Total DNA extraction generated consistent results at all oocyst levels but resulted in longer analysis time (100% and 59% detection in high- and low-inoculum-level samples, respectively). Also reflected in this study was that the matrix played an important role in the ability to recover oocysts, as sample turbidity, pH, and PCR inhibitors all influenced detection.

* Corresponding author. Mailing address: U.S. Food and Drug Administration, CFSAN/OARSA/DVA, MOD 1 Research Facility, Rm. 3603 (HFS-025), 8301 Muirkirk Rd., Laurel, MD 20708. Phone: (301) 210-7937. Fax: (301) 210-7979. E-mail: palmer.orlandi{at}fda.hhs.gov

{triangledown} Published ahead of print on 21 September 2007.

Applied and Environmental Microbiology, November 2007, p. 7474-7476, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.01652-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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