Radiation biodosimetry systems

Abstract

Disclosed herein are compositions and methods for accurately estimating the absorbed dose of radiation indicated by a subject based on the expression pattern of a panel of radiation-modulated (RM) genes at various time points following exposure of the subject to ionizing radiation.

1 . A radiation biodosimetry assay system, comprising a plurality of in vitro nucleic acid amplification reaction mixtures, each amplification reaction mixture comprising:
 (a) a probe of at least 10 nucleotides directed to at least one of SEQ ID NOs: 1-39, or the complementary sequences thereof, labeled with a fluorescent dye and a quencher; and   (b) one or more primer pairs, each primer pair comprising primers of at least 10 nucleotides directed to at least one of SEQ ID NOs: 1-39, or a complementary sequence thereof.   
     
     
         2 . The radiation biodosimetry assay system of  claim 1 , wherein the system comprises primers of at least 10 nucleotides and probes of at least 10 nucleotides directed to at least two of SEQ ID NOs: 1-39. 
     
     
         3 . The radiation biodosimetry assay system of  claim 2 , wherein the system comprises primers of at least 10 nucleotides and probes of at least 10 nucleotides directed to at least three of SEQ ID NOs: 1-39. 
     
     
         4 . The radiation biodosimetry assay system of  claim 1 , wherein the radiation biodosimetry assay system additionally comprises mRNA, or cDNA derived therefrom, from a subject exposed to ionizing radiation before the mRNA was obtained. 
     
     
         5 . The radiation biodosimetry assay system of  claim 1 , wherein the in vitro nucleic acid amplification reaction mixtures are provided in a multi-well plate. 
     
     
         6 . The radiation biodosimetry assay system of  claim 5 , wherein at least two nucleic acid probes directed to at least two different nucleic acid targets are in the same wells of the multi-well plate. 
     
     
         7 . A radiation biomarker assay kit, comprising
 a nucleic acid probe set comprising a plurality of nucleic acid probes of at least 10 nucleotides each that are directed to at least three of SEQ ID NOs: 1-39 or the complementary sequences thereof, the nucleic acid probe set further comprising a probe detectably labeled with a fluorescent dye and a quencher and configured for PCR amplification,   a set of primer pairs comprising a plurality of primer pairs each comprising primers of at least 10 nucleotides that are directed to at least three of SEQ ID NOs: 1-39 or the complementary sequences thereof, and   instructions for calculating an estimate of absorbed radiation dose from a fluorescence value obtained by contacting in vitro an mRNA sample from a human subject suspected of suffering from radiation exposure or cDNA derived therefrom to the nucleic acid probe set and a thermostable polymerase under PCR conditions.   
     
     
         8 . The radiation biomarker assay kit of  claim 7 , wherein the nucleic acid probe set comprises probes of at least 10 nucleotides directed to at least four of SEQ ID NOs: 1-39. 
     
     
         9 . The radiation biomarker assay kit of  claim 7 , wherein the nucleic acid probe set comprises probes of at least 10 nucleotides directed to at least five of SEQ ID NOs: 1-39. 
     
     
         10 . The radiation biomarker assay kit of  claim 7 , further comprising radiation exposure positive and negative control mRNA samples or cDNAs thereof. 
     
     
         11 . The radiation biomarker assay kit of  claim 7 , wherein the nucleic acid probes are provided in a multi-well plate. 
     
     
         12 . A method for estimating absorbed dose of ionizing radiation exposure received by a subject, comprising
 (i) determining the mRNA expression levels of mRNAs comprising at least one of SEQ ID NOs: 1-39 in a stabilized blood sample comprising mRNA from the subject to obtain an expression profile; and   (ii) transforming the gene expression profile into an estimate of absorbed radiation dose for the subject based on a mathematical algorithm.   
     
     
         13 . (canceled) 
     
     
         14 . (canceled) 
     
     
         15 . The method of  claim 12 , wherein transforming in step (ii) comprises calculating an estimate of absorbed radiation dose from a fluorescence value obtained by contacting in vitro an mRNA sample from a human subject suspected of suffering from radiation exposure or cDNA derived therefrom to a nucleic acid probe set and a thermostable polymerase under PCR conditions. 
     
     
         16 . The method of  claim 15 , wherein the nucleic acid probe set comprises a plurality of nucleic acid probes of at least 10 nucleotides each that are directed to at least three of SEQ ID NOs: 1-39 or the complementary sequences thereof, the nucleic acid probe set further comprising a probe detectably labeled with a fluorescent dye and a quencher and configured for PCR amplification. 
     
     
         17 . The method of  claim 15 , wherein the mRNA sample from a human subject or cDNA derived therefrom is further contacted to a set of primer pairs comprising a plurality of primer pairs each comprising primers of at least 10 nucleotides that are directed to at least three of SEQ ID NOs: 1-39 or the complementary sequences thereof. 
     
     
         18 . A method for radiation treatment triage of a subject in need thereof comprising:
 (i) determining the mRNA expression levels of mRNAs comprising at least one of SEQ ID NOs: 1-39 in a stabilized blood sample comprising mRNA from the subject to obtain a gene expression profile; and   (ii) providing a suitable treatment for radiation exposure to the subject based on the expression levels of the genes.   
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 18 , further comprising transforming the gene expression profile into an estimate of absorbed radiation dose for the subject. 
     
     
         22 . The method of  claim 21 , wherein transforming comprises calculating an estimate of absorbed radiation dose from a fluorescence value obtained by contacting in vitro an mRNA sample from a human subject suspected of suffering from radiation exposure or cDNA derived therefrom to a nucleic acid probe set and a thermostable polymerase under PCR conditions suitable for amplification of the mRNA or cDNA. 
     
     
         23 . The method of  claim 22 , wherein the nucleic acid probe set comprises a plurality of nucleic acid probes of at least 10 nucleotides each that are directed to at least three of SEQ ID NOs: 1-39 or the complementary sequences thereof, the nucleic acid probe set further comprising a probe detectably labeled with a fluorescent dye and a quencher and configured for PCR amplification. 
     
     
         24 . The method of  claim 22 , wherein the mRNA sample from a human subject or cDNA derived therefrom is further contacted to a set of primer pairs comprising a plurality of primer pairs each comprising primers of at least 10 nucleotides that are directed to at least three of SEQ ID NOs: 1-39 or the complementary sequences thereof.