US 2021/0079451 A1Published application

Isolation and detection of dna from plasma

AssigneeExact Sciences Development Company, LLC

InventorsHatim Allawi, Graham P. Lidgard, Brian Aizenstein, Tamara J. Sander, Maria Giakoumopoulos, Michael W. Kaiser, Melissa M. Gray, Abram Michael Vaccaro

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Abstract

Provided herein is technology relating to the amplification-based detection of bisulfite-treated DNAs and particularly, but not exclusively, to methods and compositions for multiplex amplification of low-level sample DNA prior to further characterization of the sample DNA. The technology further provides methods for isolating DNA from blood or blood product samples, e.g., plasma samples.

Claims21

Dependent claims

A method of processing a plasma sample, the method comprising:

a) combining a plasma sample with
i) protease;
ii) a first portion of guanidine thiocyanate;
and
iii) a first portion of non-ionic detergent,
to form a first mixture wherein proteins are digested by said protease;
b) to the first mixture of step a) adding
iv) silica particles;
v) isopropyl alcohol;
vi) a second portion of guanidine thiocyanate;
and
vii a second portion of non-ionic detergent,
to form a second mixture under conditions wherein DNA is bound to said silica particles;
c) separating silica particles with bound DNA from the second mixture;
d) to the separated silica particles with bound DNA adding a first wash solution, said first wash solution comprising guanidine hydrochloride or guanidine thiocyanate, and ethyl alcohol;
e) separating the silica particles with bound DNA from said first wash solution;
f) to the separated silica particles with bound DNA adding a second wash solution, said second wash solution comprising a buffer and ethyl alcohol;
g) separating the washed silica particles with bound DNA from said second wash solution;
h) eluting DNA from said washed silica particles with bound DNA to produce an isolated DNA sample.

16 more elements

A system for processing a plasma sample, the system comprising:

a) a first lysis reagent comprising guanidine thiocyanate and non-ionic detergent, or components for preparing said first lysis reagent;
b) a second lysis reagent comprising guanidine thiocyanate, non-ionic detergent, and isopropanol, or components for preparing said second lysis reagent;
c) a first wash solution comprising guanidine hydrochloride or guanidine thiocyanate and ethyl alcohol, or components for preparing said first wash solution;
d) a second wash solution comprising Tris buffer and ethyl alcohol, or components for preparing said second wash solution;
and
e) silica particles.

3 more elements

Dependencies are inferred from the claim text.

Facts

Application: 17/031,383Filed 24 Sep 2020
Priority: 30 Oct 2015
Published: 18 Mar 2021
Term ends: Not estimatedThe source records don’t support an estimate.

Classification8 codes

C12Q 1/6806
Involving nucleic acids: preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12N 15/1006
Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses…
C12Q 1/6837
Enzymatic or biochemical coupling of nucleic acids to a solid phase: using probe arrays or probe chips
C12Q 1/686
Nucleic acid amplification reactions: polymerase chain reaction [PCR]

Family

Application62/249,097Us provisional application · filed 30 Oct 2015
Application15/335,111Continuation · filed 26 Oct 2016
Application17/031,383Continuation
US 10,822,638 B2Related patent
US 11,674,168 B2Related patent
US 2017/0121704 A1Published application
US 2021/0079451 A1This patent

From the US filing history. Foreign family members aren’t listed.

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