WHOLE GENOME TARGETED ENRICHMENT AND SEQUENCING OF HUMAN-INFECTINGCRYPTOSPORIDIUMspp.

Abstract

AbstractCryptosporidiumspp. are protozoan parasites that cause severe illness in vulnerable human populations. Obtaining pureCryptosporidiumDNA from clinical and environmental samples is challenging because the oocysts shed in contaminated feces are limited in quantity, difficult to purify efficiently, may derive from multiple species, and yield limited DNA (<40 fg/oocyst). Here, we develop and validate a set of 100,000 RNA baits (CryptoCap_100k) based on six human-infectingCryptosporidiumspp. (C. cuniculus,C. hominis,C. meleagridis,C. parvum,C. tyzzeri, andC. viatorum) to enrichCryptosporidiumspp. DNA from a wide array of samples. We demonstrate that CryptoCap_100k increases the percentage of reads mapping to targetCryptosporidiumreferences in a wide variety of scenarios, increasing the depth and breadth of genome coverage, facilitating increased accuracy of detecting and analyzing species within a given sample, while simultaneously decreasing costs, thereby opening new opportunities to understand the complex biology of these important pathogens.