US 12,123,051 B2
Nucleic acid ratio determination
Alison Velyian Todd, Glebe (AU); and Nicole Elizabeth Lima, Cremorne (AU)
Assigned to SPEEDX PTY LTD, Eveleigh (AU)
Appl. No. 16/619,932
Filed by SpeeDx Pty Ltd, Eveleigh (AU)
PCT Filed Dec. 21, 2018, PCT No. PCT/AU2018/051406
§ 371(c)(1), (2) Date Dec. 5, 2019,
PCT Pub. No. WO2019/119072, PCT Pub. Date Jun. 27, 2019.
Claims priority of application No. 2017905138 (AU), filed on Dec. 21, 2017.
Prior Publication US 2020/0199651 A1, Jun. 25, 2020
Int. Cl. C12Q 1/686 (2018.01); C12Q 1/18 (2006.01); C12Q 1/6851 (2018.01); G16B 20/00 (2019.01); G16B 25/10 (2019.01); G16B 25/20 (2019.01)
CPC C12Q 1/686 (2013.01) [C12Q 1/18 (2013.01); C12Q 1/6851 (2013.01); G16B 20/00 (2019.02); G16B 25/10 (2019.02); G16B 25/20 (2019.02)] 20 Claims
 
1. A method for normalising quantitative data obtained by amplification of nucleic acids from a cell, organism, or virus, the method comprising:
(i) performing a combined amplification of genomic DNA from a first gene and RNA transcribed from the first gene thereby obtaining quantitative data, which is a combined measure of the genomic DNA from the first gene and RNA transcribed from the first gene, and an amplification of a sequence of non-transcribed genomic DNA in the cell organism or virus, thereby obtaining quantitative data, which is a measure of the sequence of non-transcribed genomic DNA,
wherein the genomic DNA from the first gene and RNA transcribed from the first gene, and the sequence of non-transcribed genomic DNA are co-amplified in the same reaction,
wherein the nucleic acids from the cell, organism, or virus are an extract of total nucleic acids,
and wherein the reaction comprises using reverse transcriptase;
(ii) normalizing the quantitative data from the combined amplification to the quantitative data from the amplification of the sequence of the non-transcribed genomic DNA to derive a normalised value (nV) representative of the relative amounts of:
said genomic DNA and RNA transcripts of the first gene, to the non-transcribed genomic DNA, present within the nucleic acid sample prior said amplification.