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US 11,293,049 B2Granted
Modulation of accessibility of host nucleic acids to nucleic acid digesting enzymes in acellular biological fluids
AssigneePathoquest
InventorÉric Cabannes
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Abstract
Disclosed is a method for isolating, amplifying, and sequencing infectious agents' nucleic acids from an acellular fraction of a biological fluid using detergents and nucleic acids-digesting enzymes. Also disclosed is a kit-of-parts including detergents and nucleic acids-digesting enzymes for the implementation of the methods described.
Claims10
A method for isolating, amplifying, and sequencing viral nucleic acids from a biological fluid comprising:
- a) centrifuging the biological fluid under conditions suitable for ensuring the elimination of cells, vesicles and aggregates in the resulting supernatant, and recovering said supernatant, thereby obtaining a viral fraction of said biological fluid,
- b) contacting the viral fraction of the biological fluid with at least one detergent and at least one nucleic acids-digesting enzyme,
- c) inactivating the at least one nucleic acids-digesting enzyme,
- d) extracting viral nucleic acids,
- e) optionally, reverse-transcribing viral genomic RNAs,
- f) amplifying viral genomic DNA and/or, where viral genomic RNAs were reverse-transcribed at step e), viral cDNA, and
- g) sequencing amplified viral genomic DNA and/or, where viral genomic RNAs were reverse-transcribed at step e), amplified viral cDNA.
4 more elements
Dependencies are inferred from the claim text.