Preparation of dna sequencing libraries for detection of dna pathogens in plasma
AssigneeIllumina, Inc.
InventorsTong Liu, Fiona Kaper, Clifford Wang
Life of this patent
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Abstract
The application provides an agnostic, shotgun nucleic acid sequencing-based method for the detection of pathogens in samples from human patients, animals, or plants. The method includes dehosting the sample of the nucleic acid molecules of host origin and provides for the detection of pathogens without prior knowledge of their genome sequences.
Claims23
A sample preparation method comprising:
- obtaining a host organism sample;
- removing intact cells from the host organism sample;
- removing nucleic acid molecules of less than 1000 basepairs (bp) from the host organism sample to obtain a dehosted sample.
A method of dehosting a sample obtained from a host organism, the method comprising:
- removing intact cells from the host organism sample;
- removing nucleotide acid molecules of less than 1000 basepairs (bp) from the host organism sample to obtain a dehosted sample.
A method of identifying pathogen nucleotide sequences in a sample obtained from a host organism, the method comprising:
- removing intact cells from the host organism sample;
- removing nucleotide acid molecules of less than 1000 basepairs (bp) from the host organism sample to obtain a dehosted sample;
- preparing a sequencing library from the nucleic acid molecules remaining in the dehosted sample;
- sequencing the nucleotide sequences of the sequencing library;
- and
- identifying pathogen sequences within the sequenced sequences.
The method of 1, wherein removing nucleotide acid molecules of less than 1000 bp from the host organism sample comprises solid phase reversible immobilization (SPRI) beads under conditions favoring capture of nucleotide molecules of 1000 bp or greater.
Dependencies are inferred from the claim text.