Development of Procedures for Direct Extraction of Cryptosporidium.pdfVIP

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Mar. 2005, p. 1135–1141 Vol. 71, No. 3 0099-2240/05/$08.00 0 doi:10.1128/AEM.71.3.1135–1141.2005 Copyright © 2005, American Society for Microbiology. All Rights Reserved. Development of Procedures for Direct Extraction of Cryptosporidium DNA from Water Concentrates and for Relief of PCR Inhibitors Jianlin Jiang,1 2 3 1 Kerri A. Alderisio, Ajaib Singh, and Lihua Xiao * Division of Parasitic Disease, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 1 2 ; New York City Department of Environmental Protection, Valhalla, New York ; and City of Milwaukee Public Health Laboratories, Milwaukee, Wisconsin3 Received 16 July 2004/Accepted 7 October 2004 Extraction of high-quality DNA is a key step in PCR detection of Cryptosporidium and other pathogens in environmental samples. Currently, Cryptosporidium oocysts in water samples have to be purified from water concentrates before DNA is extracted. This study compared the effectiveness of six DNA extraction methods (DNA extraction with the QIAamp DNA minikit after oocyst purification with immunomagnetic separation and direct DNA extraction methods using the FastDNA SPIN kit for soil, QIAamp DNA stool minikit, UltraClean soil kit, or QIAamp DNA minikit and the traditional phenol-chloroform technique) for the detection of Cryptosporidium with oocyst-seeded s