Quantifying C. parvum Oocyst Shedding

Analyses by qRT-PCR with fecal and ileal C. parvum oocyst shedding were assessed by genomic DNA extracted from fecal and ileal samples. The primers target the 18 s rRNA gene of the parasite by GenBank (www.ncbi.nlm.nih.gov/genbank/, AF164102). The reaction was performed in a Bio-Rad iCycler iQ multicolor PCR Detection System using iCycler software (version 3.0). Amplification consisted of 15 min at 95 °C followed by 40 cycles of 15 s at 95 °C, 15 s at 52 °C, and 20 s at 72 °C, followed by 0.5-degree increments for 10 s starting at 75 °C and ending with 95 °C for the Melt Curve. Ct values of each run were compared to standards with known amounts of C. parvum DNA and log transformed into number of organisms per mg of stool sample.¹⁹ (link)