Non-invasive in-vitro method for somatic mutation detection

Abstract

The invention relates to an in-vitro method of detecting a somatic mutation in a human comprising the steps of, (i) providing for a human sample, preferably a blood sample from a subject, more preferably a plasma sample, or a serum sample or a buffy coat sample, (ii) preparing a first nucleic acid sequencing library from the nucleic acids present in said sample, (iii) hybridizing one or more TAC oligonucleotides from a first TAC oligonucleotide pool (TAC oligonucleotide-1) to said first nucleic acid library thereby isolating a first subset of library nucleic acid molecules, (iv) sequencing said first subset of library nucleic acid molecules and comparing the sequences determined to a human reference sequence, thereby creating a first group of putatively informative sequence variants (PISV1), (v) hybridizing one or more TAC oligonucleotides from a second smaller TAC oligonucleotide pool comprising TAC oligonucleotides (TAC oligonucleotide-2) which are specific for the nucleic acid molecules of said first group of putatively informative sequence variants, to said first library of nucleic acid molecules thereby isolating a second subset of library nucleic acid molecules, (vi) sequencing said second subset of library nucleic acid molecules and comparing the sequences determined to a human reference sequence, thereby creating a second group of putatively informative sequence variants (PISV2), (vii) analyzing the putatively informative sequence variants (PISV2) thereby detecting a somatic mutation in the sample from the subject. Steps (i) to (iv) are also referred to together as step A. Steps (v) to (vii) are also referred to together as step B.

Dependencies are inferred from the claim text.