Device and method for multiple analyte detection
View Patent ↗The invention is directed to a method and device for simultaneously testing a sample for the presence, absence, and/or amounts of one or more of a plurality of selected analytes. The invention includes, in one aspect, a device for detecting or quantitating a plurality of different analytes in a liquid sample. Each chamber may include an analyte-specific reagent effective to react with a selected analyte that may be present in the sample, and detection means for detecting the signal. Also disclosed are methods utilizing the device.
1. A method for reacting a target nucleic acid and detecting an optical signal, comprising:
providing a substrate comprising a chamber, wherein the chamber is pre-loaded with a first unlabeled oligonucleotide such that the first unlabeled oligonucleotide is immobilized on the substrate, and the chamber is pre-loaded with a second unlabeled oligonucleotide such that the second unlabeled oligonucleotide is immobilized on the substrate;
introducing a target nucleic acid sequence into the chamber;
reacting the target nucleic acid sequence with both the first unlabeled oligonucleotide and the second unlabeled oligonucleotide in the chamber, to form a first reaction product in the chamber;
reacting the first reaction product with an optically detectable detection reagent in the chamber to form a second reaction product; and
detecting an optical signal produced in the chamber by the second reaction product in the chamber,
wherein the substrate further comprises a channel in fluid communication with the chamber.
2. The method of claim 1 , wherein reacting the target nucleic acid sequence with both the first unlabeled oligonucleotide and the second unlabeled oligonucleotide in the chamber, to form a first reaction product in the chamber, comprises at least one of a polymerase chain reaction, a ligase chain reaction, an oligonucleotide ligation assay, and a hybridization assay.
3. The method of claim 1 , wherein the first unlabeled oligonucleotide and the second unlabeled oligonucleotide comprise a primer pair suitable for amplifying the target nucleic acid sequence.
4. The method of claim 1 , wherein at least one of the first unlabeled oligonucleotide and the second unlabeled oligonucleotide comprises a sequence that is complementary to at least a portion of the target nucleic acid sequence.
5. The method of claim 1 , wherein the second reaction product is detected through an optically transparent window in the chamber.
6. The method of claim 1 , wherein the target nucleic acid sequence is introduced into the chamber by vacuum action.
7. The method of claim 1 , wherein at least one of the pre-loaded first unlabelled oligonucleotide and the pre-loaded second unlabelled oligonucleotide are dried in the chamber.
8. The method of claim 1 , wherein the optically detectable detection reagent comprises a fluorescent dye.
9. The method of claim 1 , wherein the optically detectable detection reagent comprises a fluorescer dye and a quencher dye.
10. The method of claim 1 , wherein at least one of the first unlabeled oligonucleotide and the second unlabeled oligonucleotide comprises a polynucleotide analog.
11. The method of claim 1 , wherein the first unlabeled oligonucleotide and the second unlabeled oligonucleotide bind to adjacent, contiguous regions of the target nucleic acid sequence.
12. The method of claim 1 , wherein the first unlabeled oligonucleotide has a sequence that is complementary to at least a portion of the target nucleic acid sequence.
13. The method of claim 1 , wherein the second unlabeled oligonucleotide has a sequence that is complementary to at least a portion of the target nucleic acid sequence.
14. The method of claim 1 , wherein the first unlabeled oligonucleotide has a sequence that is complementary to at least a first portion of the target nucleic acid sequence, and where the second unlabeled oligonucleotide has a sequence that is complementary to at least a second portion of the target nucleic acid sequence.
15. The method of claim 1 , wherein the reacting of the target nucleic acid sequence with both the first unlabeled oligonucleotide and the second unlabeled oligonucleotide in the chamber, further comprises reacting the target nucleic acid sequence with nucleotide triphosphates, to form the first reaction product in the chamber.
16. The method of claim 1 , wherein the chamber comprises a plurality of chambers formed in the substrate, the channel comprises a plurality of channels, and each of the plurality of chambers is in fluid communication with one of the plurality of channels.