Quantifying Replication Slippage Error in Cryptosporidium Metabarcoding Studies.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 39052741.
- Also identified by DOI 10.1093/infdis/jiae065 and PMC identifier 11272095.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Genetic variation in Cryptosporidium, a common protozoan gut parasite in humans, is often based on marker genes containing trinucleotide repeats, which differentiate subtypes and track outbreaks. However, repeat regions have high replication slippage rates, making it difficult to discern biological diversity from error. Here, we synthesized Cryptosporidium DNA in clonal plasmid vectors, amplified them in different mock community ratios, and sequenced them using next-generation sequencing to determine the rate of replication slippage with dada2. Our results indicate that slippage rates increase with the length of the repeat region and can contribute to error rates of up to 20%.
Medical subject headings
- Cryptosporidium
- DNA Replication