Genetic analysis and authentication
View Patent ↗This invention provides compositions and methods for genetic testing of an organism and for correlating the results of the genetic testing with a unique marker that unambiguously identifies the organism. The markers may be internal markers, such as for example single nucleotide polymorphisms (SNPs), short tandem repeats (STRs), or other sites within a genomic locus. Alternatively, the markers may be external, such that they are separately added to the genetic sample before testing.
1. A method for analyzing a target nucleic acid sequence in a patient genomic sample and for identifying said patient providing the sample, said method comprising the following steps:
(a) providing a first set of oligonucleotides, wherein members of the first set have different sequences and said members are attached to beads which are associated with an optically distinguishable characteristic that identifies the sequences of the oligonucleotides attached thereto, the first set of oligonucleotides for identifying target nucleic acid sequences in the patient sample;
(b) a second set of oligonucleotide wherein members of the second set have different sequences and said members are attached to beads which are associated with an optically distinguishable characteristic that identities the sequences of the oligonucleotides attached thereto, the second set of oligonucleotides for identifying a number of marker sequences in the patient genomic sample, said marker sequences each including at least one polymorphic marker;
(c) contacting the target sequence and a plurality of marker sequences to said first and said second set of oligonucleotides under conditions permitting annealing of the first set of oligonucleotides to the target sequence or annealing of the second set of oligonucleotides to the marker sequences, or said annealing followed by elongation of the annealed oligonucleotides; and
(d) detecting the annealing or annealing followed by elongation between the oligonucleotides of the first set to the target sequence and the oligonucleotides of the second set with the marker sequences, said method thereby providing the analysis of the target sequence and the identification of said patient.
2. The method of claim 1 , wherein the beads are arranged in a planar array.
3. The method of claim 2 , wherein the bead array composes subarrays, with the oligonucleotides of the second set and oligonucleotides of the first set being located in different subarrays.
4. The method of claim 2 , wherein the planar bead array is disposed on an electrode.
5. A method for analyzing a target nucleic acid sequence in a patient genomic sample and for identifying said patient providing the sample, said method comprising the following steps:
(a) providing a first set of oligonucleotides wherein members of the first set have different sequences and sad members are attached to beads which are associated with an optically distinguishable characteristic that identifies the sequences of the oligonucleotides attached thereto, the first set of oligonucleotides for identifying target nucleic acid sequences;
(b) contacting said first set of oligonucleotides with a solution containing the target nucleic acid sequence under conditions permitting annealing of the first set of oligonucleotides with the target nucleic acid sequence or said annealing followed by elongation of the annealed oligonucleotides;
(c) labeling the solution with a molecular label that uniquely identifies said target solution, such the patient identity is determined by interrogating said label, wherein the label may be added to said sample before said target solution is contacted with the first set of oligonucleotides, or afterwards, or concurrently therewith; and
(d) detecting said annealing or said annealing followed by elongation of said oligonucleotides and correlating the optically distinguishable characteristic with the oligonucleotides which annealed or which were elongated in order to identify said oligonucleotides and thereby identify the patient providing the sample.
6. The method of claim 2 , wherein the beads are associated with a chemical label that uniquely identifies the oligonucleotides attached to said beads, wherein the chemical label comprises one or more fluorophore dyes.
7. The method of claim 2 , wherein the target nucleic acid sequence is analyzed to determine if it contains a mutation and wherein the target nucleic acid and the polymorphic markers are located on the same gene.
8. The method of claim 2 , wherein the target nucleic acid sequence is analyzed to determine if it contains a mutation and wherein the target nucleic acid and the polymorphic markers are located in different genes.
9. The method of claim 5 , further comprising the step of detecting the molecular label to identity the patient.
10. The method of claim 5 , wherein the labeling of the sample comprises adding one or more fluorescent tags, said tags thereby becoming associated with the target solution.
11. The method of claim 5 , wherein the labeling of the target solution comprises adding oligonucleotide sequence tags.
12. The method of claim 11 , further comprising a set of sequence tag probes designed to hybridize with the sequence tags, said sequence tag probes being attached to beads, wherein said beads are encoded to uniquely identify the sequence tag probes attached to said beads.
13. The method of claim 1 wherein the target nucleic acid sequence and the marker sequences are is derived from the patient genomic sample by amplification of one or more nucleic acid subsequences within the patient genomic sample.
14. The method of claim 5 wherein the target nucleic acid sequence is derived from the patient genomic sample by amplification of one or more nucleic acid subsequences within the patient genomic sample.
15. The method of claims 13 or 14 wherein the amplification is performed using the polymerase chain reaction.
16. The method of claim 1 or 5 wherein the oligonucleotides are capable of being elongated following annealing, by incorporation of one or more types of deoxyribonucleotide triphosphates or di-deoxyribonucleotide triphosphates.
17. The method of claim 1 or 5 , wherein the target nucleic acid sequence includes a mutation site and oligonucleotides in the set are designed to detect different nucleotides at the mutation site.
18. The method of claim 1 , wherein the marker sequence includes a polymorphic site and said second set includes members designed to detect the different nucleotides at the polymorphic site.
19. The method of claim 1 or 5 , wherein the target sequence includes a mutation site and oligonucleotides in the set are capable of annealing to subsequences in the target sequence including the mutation site, or within a range of proximity to the mutation site.
20. The method of claim 1 , wherein the marker sequence includes a polymorphic site and the second set of oligonucleotides includes members capable of annealing to subsequences in the marker sequence including the polymorphic site, or within a range of proximity to the polymorphic site.
21. The method of claims 19 , wherein the mutation or polymorphic site is a single nucleotide polymorphism.
22. The method of claim 19 , wherein the mutation or polymorphic site is a short term tandem repeat site.
23. The method of claim 16 wherein a fraction of at least one type of deoxyribonucleotide triphosphate or di-deoxyribonucleotide triphosphate is labeled so as to generate an optically detectable signature associated with the elongation product following incorporation.
24. The method of claim 23 wherein the labels are fluorophore dyes.
25. The method of claim 1 or 5 wherein the label is an inorganic nanoparticle label, including semiconductor Quantum Dot particles and Resonant Light Scattering metal nanoparticles.