US 12,305,235 B2
Methods for detecting immune cell DNA and monitoring immune system
Dina M. Hafez, Los Altos, CA (US); Prashanthi Natarajan, Foster City, CA (US); Raheleh Salari, San Carlos, CA (US); Ryan Swenerton, Millbrae, CA (US); and Bernhard Zimmermann, Manteca, CA (US)
Assigned to Natera, Inc., San Carlos, CA (US)
Appl. No. 17/616,536
Filed by Natera, Inc., San Carlos, CA (US)
PCT Filed May 29, 2020, PCT No. PCT/US2020/035216
§ 371(c)(1), (2) Date Dec. 3, 2021,
PCT Pub. No. WO2020/247263, PCT Pub. Date Dec. 10, 2020.
Claims priority of provisional application 62/857,966, filed on Jun. 6, 2019.
Prior Publication US 2022/0251654 A1, Aug. 11, 2022
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6881 (2018.01); C12Q 1/6844 (2018.01)
CPC C12Q 1/6881 (2013.01) [C12Q 1/6844 (2013.01); C12Q 2600/156 (2013.01); C12Q 2600/16 (2013.01)] 16 Claims
OG exemplary drawing
 
1. A method of preparing a non-naturally occurring composition of amplified DNA derived from a biological sample useful for detecting or monitoring immune cells in a subject, comprising:
preparing a non-naturally occurring composition of amplified DNA by performing a multiplex amplification reaction on nucleic acids isolated from a biological sample of the subject to generate a set of amplicons, wherein each of the set of amplicons comprises recombined V(D)J gene segments at a gene locus of interest, wherein the multiplex amplification reaction is capable of amplifying at least about 70% of all possible V(D)J recombinations at the gene locus of interest, wherein the multiplex amplification reaction is performed with a first set of primers targeting V gene sequences targeted by one or more of the primers of SEQ ID NOs: 1-64 and a second set of primers targeting J gene sequences targeted by one or more of the primers of SEQ ID Nos: 65-76; and
analyzing the non-naturally occurring composition of amplified DNA by sequencing the set of amplicons, wherein sequences of the recombined V(D)J gene segments are indicative of presence of an immune cell in the biological sample.