Fast and reliable extraction of protozoan parasite DNA from fecal specimens

Background: Polymerase chain reaction (PCR) detection of intestinal protozo a in fecal specimens is hampered by poor recovery of DNA and by the presenc e of PCR inhibitors. In this study we describe a novel method for DNA extra ction from such specimens containing spores and oocysts of Enterocytozoon b ieneusi and Cryptosporidium parvum, respectively. Methods and Results: Extraction was done using commercial kits modified to maximize the recovery and purity of extracted DNA. In comparison with a pro cedure we previously reported, we estimate that this method may increase th e sensitivity of parasite DNA detection in fecal specimens up to tenfold. A n additional advantage of this method is that up to 12 samples may be proce ssed simultaneously within 2 hours. Conclusions: By using this method, we were able to increase reproducibility of PCR amplification on fecal specimens and significantly reduce the hands -on time required to process the samples.