REACTION MIXTURES FOR DETECTING NUCLEIC ACIDS ALTERED BY CANCER IN PERIPHERAL BLOOD
Analyzing peripheral blood RNA populations presents an effective, accurate, minimally invasive method of determining a patient's cancer status. Using circulating free RNA of the genes disclosed herein, systems and methods are disclosed which can accurately identify cancer signatures in the patient blood samples.
1 . A reaction mixture comprising:
a biological sample suspected of containing circulating free nucleic acids derived from a target formylpeptide receptor gene;
one or more oligonucleotide probes specific for the circulating free nucleic; and
amplification reagents for amplifying the circulating free nucleic acids using the one or more oligonucleotide probes.
2 . The reaction mixture of claim 1 , wherein the target formylpeptide receptor gene is FPR1.
3 . The reaction mixture of claim 1 , wherein the biological sample comprises blood components.
4 . The reaction mixture of claim 1 , wherein the circulating free nucleic acids comprises circulating free RNA transcripts derived from the target formylpeptide receptor gene.
5 . The reaction mixture of claim 1 , wherein the one or more oligonucleotide probes comprise an amplification primer, a sequencing primer, a microarray probe, or a combination of any thereof.
6 . The reaction mixture of claim 1 , wherein at least one of the one or more oligonucleotide probes comprises a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
7 . The reaction mixture of claim 1 , wherein the amplification reagents comprise one or more of the following reagents: nucleic acid polymerases, reverse transcriptases, DNAses, RNases, reaction buffers, reducing agents, surfactants, divalent cations, and dNTPs.
8 . The reaction mixture of claim 1 , wherein the reaction mixture is a qPCR mixture reaction or an RT-PCR mixture reaction.
9 . A kit for determining cancer risk in a subject, comprising one or more oligonucleotide probes specific for circulating free nucleic acids derived from a formylpeptide receptor gene and a quantification standard for quantifying the level of circulating free formylpeptide receptor nucleic acids in, the subject.
10 . The kit of claim 9 , wherein the quantification standard comprises synthetic formylpeptide receptor nucleic acids.
11 . The kit of claim 9 , wherein the quantification standard comprises at least three synthetic nucleic acid molecules having the nucleotide sequences set forth in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively.
12 . The kit of claim 9 , further comprising oligonucleotide probes specific for circulating free nucleic acids derived from one or more reference genes.
13 . The kit of claim 12 , wherein the one or more reference genes comprise an Actin B gene (ACTB) or a heterogeneous nuclear ribonucleoprotein gene (HNRNPA1).
14 . The kit of claim 13 , wherein at least one of the one or more probes specific for circulating free nucleic acids derived from ACTB comprises a nucleotide sequence of SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6.
15 . The kit of claim 13 , wherein at least one of the one or more probes specific for circulating free nucleic acids derived from HNRNPA1 comprises a nucleotide sequence of SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9.
16 . The kit of claim 8 , wherein, the kit is configured to determine lung cancer risk.
17 . The kit of claim 16 , wherein the kit is configured to determine non-small cell lung cancer risk.
18 . The kit of claim 8 , further comprising reagents suitable for the lysis of cells.
19 . The kit of claim 9 , further comprising reagents for the isolation of circulating free nucleic acids, and/or for the storage of the circulating free nucleic acids.
20 . The kit of claim 9 , further comprising a notice, in the form prescribed by a governmental agency regulating the manufacture, sale, or use of pharmaceutical or biological products, which reflects approval by the agency of manufacture, sale, or use for human administration.