Radiation biodosimetry systems

Patent metadata

Field	Value
Publication number	US-10435747-B2
Application number	US-201514823433-A
Country	US
Kind code	B2
Filing date	Aug 11, 2015
Priority date	Aug 19, 2014
Publication date	Oct 8, 2019
Grant date	Oct 8, 2019

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.

First claim

We claim:

1. A radiation biodosimetry assay system, comprising a plurality of in vitro nucleic acid amplification reaction mixtures, each amplification reaction mixture comprising:
(i) a probe of at least 10 nucleotides directed to SEQ ID NOs: 6, 8, 9, 10 or 14, or the complementary sequence thereof, labeled with a fluorescent dye and a quencher; and
(ii) one or more primer pairs, each primer pair comprising primers of at least 10 nucleotides directed to SEQ ID NOs: 6, 8, 9, 10, or 14, or a complementary sequence thereof, wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to at least three of SEQ ID NOs: 6, 8, 9, 10, or 14, or the complementary sequences thereof.

2. The radiation biodosimetry assay system of claim 1 , wherein the radiation biodosimetry assay system additionally comprises mRNA or cDNA from a subject exposed to ionizing radiation before the mRNA or cDNA was obtained.

3. The radiation biodosimetry assay system of claim 1 , wherein the in vitro nucleic acid amplification reaction mixtures are provided in a multi-well plate.

4. The radiation biodosimetry assay system of claim 3 , 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.

5. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs directed to at least four of SEQ ID NOs: 6, 8, 9, 10, or 14, or complementary sequences thereof.

6. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to SEQ ID NOs: 6, 8, and 10, or the complementary sequences thereof.

7. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to SEQ ID NOs: 6, 8, and 14, the complementary sequences thereof.

8. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to SEQ ID NOs: 8, 9, and 10, or the complementary sequences thereof.

9. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to SEQ ID NOs: 8, 9, and 14, or the complementary sequences thereof.

10. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to SEQ ID NOs: 6, 9, and 10, or the complementary sequences thereof.

11. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to SEQ ID NOs: 6, 9, and 14, or the complementary sequences thereof.

12. The radiation biodosimetry assay system of claim 1 , wherein the system comprises primer pairs of at least 10 nucleotides and probes of at least 10 nucleotides directed to SEQ ID NOs: 6, 8, and 9, or the complementary sequences thereof.

13. 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: 6, 8, 9, 10, or 14 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: 6, 8, 9, 10, or 14 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 or cDNA sample from a human subject suspected of suffering from radiation exposure to the nucleic acid probe set and a thermostable polymerase under PCR conditions.

14. The radiation biomarker assay kit of claim 13 , wherein the nucleic acid probe set comprises no more than 100 probes.

15. The radiation biomarker assay kit of claim 13 , wherein the nucleic acid probe set comprises nucleic acid probes directed to each of SEQ ID NOs: 6, 8, 9, 10, and 14 or the complementary sequences thereof.

16. The radiation biomarker assay kit of claim 13 , further comprising radiation exposure positive and negative control mRNA samples or cDNAs thereof.

17. The radiation biomarker assay kit of claim 13 , further comprising a nucleic acid probe of at least 10 nucleotides directed to at least one of SEQ. ID NOs:14, 16, 17, 25, or 29, or the complementary sequences thereof.

18. The radiation biomarker assay kit of claim 17 , wherein the probe set comprises no more than 100 probes.

19. The radiation biomarker assay kit of claim 17 , wherein the nucleic acid probes are provided in a multi-well plate.

20. The radiation biomarker assay kit of claim 19 , 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.

21. The radiation biomarker assay kit of claim 17 , further comprising radiation exposure positive and negative control mRNA samples or cDNAs thereof.

22. The radiation biomarker assay kit of claim 13 , wherein the kit comprises nucleic acid probes directed to at least four of SEQ ID NOs: 6, 8, 9, 10, or 14, or complementary sequences thereof.

Assignees

• Labaer Joshua
• Gillis Kristin
• Wallstrom Garrick
• Park Jin
• Murugan Vel
• Magee Mitch
• Univ Arizona State

Inventors

• Labaer Joshua
• Gillis Kristin
• Wallstrom Garrick
• Park Jin
• Murugan Vel
• Magee Mitch

Classifications

• C12Q1/6827

  for detection of mutation or polymorphism · CPC title

• C12Q2600/156

  Polymorphic or mutational markers · CPC title

• C12Q1/6883 Primary

  for diseases caused by alterations of genetic material · CPC title

• C12Q2537/165

  Mathematical modelling, e.g. logarithm, ratio · CPC title