Single Oocyst Selection and Whole-Genome Amplification of Cryptosporidium parvum.

Cryptosporidium parvum is a zoonotic parasite of the intestine and poses a threat to human and animal health. However, it is difficult to obtain a large number of oocysts for genome sequencing using in vitro culture. To address this challenge, we describe a procedure for purification of C. parvum from a clinically positive sample and a method for isolation of a single oocyst of C. parvum using a three-axis hydraulic micromanipulator. We employ the strategy of whole-genome amplification (WGA) of 10 oocysts, and purify the whole-genome DNA after amplification, and finally meet the basic conditions for long-read sequencing of C. parvum.