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Quantifying Replication Slippage Error in Cryptosporidium Metabarcoding Studies.

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%.

SUBMITTER: Knox MA

PROVIDER: S-EPMC11272095 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature

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Quantifying Replication Slippage Error in Cryptosporidium Metabarcoding Studies.

Knox Matthew A MA Biggs Patrick J PJ Garcia-R Juan Carlos JC Hayman David T S DTS

The Journal of infectious diseases 20240701 1

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 dete ...[more]

PMID: 39052741

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