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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.

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Single Oocyst Selection and Whole-Genome Amplification of Cryptosporidium parvum

  • 实验方案

  • 发表日期：

    2026年

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  • Yuancai Chen,
  • Kaihui Zhang,
  • Jianying Huang,
  • Huikai Qin,
  • Zi Yan,
  • Yin Fu,
  • Junqiang Li,
  • Rongjun Wang,
  • Longxian Zhang

摘要

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.