Reaction mixtures for detecting nucleic acids altered by cancer in peripheral blood

Abstract

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.

What is claimed is: 
     
         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.