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US 10,870,891 B2Granted
Diagnostic test system for specific, sensitive and reproducible detection of circulating nucleic acids in whole blood
AssigneeBiodesix, Inc.
InventorsHestia Mellert, Leisa Jackson, Gary A. Pestano
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Abstract
The present disclosure relates to a rapid diagnostic test system that includes the prospective collection of whole blood, preservation of circulating nucleic acids at ambient temperature, and the reproducible detection of nucleic acids including DNA and mRNA (including fusion transcripts and differentially expressed transcripts) by different genomic methodologies.
Claims15
A method of detecting a fragmented RNA in a freshly-collected whole blood sample from a mammalian subject comprising:
- a) recovering blood components from a whole blood sample from a mammalian subject, said blood components including plasma and buffy coat from the whole blood sample;
- b) isolating free RNA, RNA associated with, exosomes and/or platelets in platelet enriched plasma (PEP), RNA associated with PBMCs, or a combination thereof from said blood components;
- c) purifying and concentrating the isolated RNA of step (b);
- d) amplifying said RNA of step (c) or a complimentary DNA generated from said RNA of step (c) wherein generating complimentary DNA comprises performing reverse transcription of said isolated RNA using SuperScript IV reverse transcriptase;
- e) purifying and concentrating the amplified RNA;
- and
- f) identifying and/or quantifying amplified said RNA or complimentary DNA from step (e).
4 more elements
A method of detecting a cytokeratin nucleic acid in a sample comprising:
- a) obtaining a cytokeratin RNA containing sample;
- b) annealing forward and reverse primers that hybridize to said cytokeratin nucleic acid with said sample;
- c) amplifying said cytokeratin RNA;
- and
- d) detecting amplified cytokeratin RNA.
2 more elements
Dependencies are inferred from the claim text.