CRISPR-Cas9-mediated host signal reduction for 18S metabarcoding of host-associated eukaryotes.
Mol Ecol Resour
; 24(6): e13980, 2024 Aug.
Article
em En
| MEDLINE
| ID: mdl-38804043
ABSTRACT
Metabarcoding-based methods for identification of host-associated eukaryotes have the potential to revolutionize parasitology and microbial ecology, yet significant technical challenges remain. In particular, highly abundant host reads can mask the presence of less-abundant target organisms, especially for sample types rich in host DNA (e.g., blood and tissues). Here, we present a new CRISPR-Cas9-mediated approach designed to reduce host signal by selective amplicon digestion, thus enriching clinical samples for eukaryotic endosymbiont sequences during metabarcoding. Our method achieves a nearly 76% increased efficiency in host signal reduction compared with no treatment and a nearly 60% increased efficiency in host signal reduction compared with the most commonly used published method. Furthermore, the application of our method to clinical samples allows for the detection of parasite infections that would otherwise have been missed.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
RNA Ribossômico 18S
/
Eucariotos
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Código de Barras de DNA Taxonômico
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Sistemas CRISPR-Cas
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Mol Ecol Resour
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Estados Unidos