Rapid methods to isolate Cryptosporidium DNA from frozen feces for PCR.
The polymerase chain reaction (PCR) offers one of the most sensitive methods for detecting Cryptosporidium parvum but its sensitivity in fecal material can be greatly reduced by a variety of poorly defined 'inhibitors'. Methods that separate the parasitic oocyst from fecal material prior to DNA extraction circumvent inhibitor interference but are problematic for frozen specimens since 'intact' oocysts are required for separation. We report here a relatively low-cost, rapid method for extracting C. parvum DNA from frozen fecal materials that can be used in a PCR assay for detection of single parasitic oocysts.
Ward
L A
LA
The Ohio State University, Ohio Agricultural Research and Development Center, Department of Veterinary Preventive Medicine and Food Animal Health Research Program, Wooster, Ohio 44691, USA. ward.195@osu.edu
Wang
Y
Y
D000818:Animals; D015925:Cryopreservation; D003458:Cryptosporidium / Q000235:genetics*; D016054:DNA, Protozoan / Q000302:isolation & purification*; D005243:Feces / Q000469:parasitology*; D006801:Humans; D007124:Immunoenzyme Techniques; D016133:Polymerase Chain Reaction
D016054:DNA, Protozoan