Device and method for multiple analyte detection
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 comprising:
pre-loading a first unlabeled oligonucleotide into a chamber;
pre-loading a second unlabeled oligonucleotide into the chamber;
after the pre-loading of the first unlabeled oligonucleotide and of the second unlabeled oligonucleotide, introducing a sample comprising a target sequence into the chamber;
reacting the target sequence with both the first unlabeled oligonucleotide and the second unlabeled oligonucleotide in the chamber, to form a reaction product in the chamber;
producing a detectable signal from the reaction product in the chamber; and
detecting more than one optical signal indicative of more than one different analyte in the sample, wherein the more than one optical signal comprises the detectable signal.
2 . The method of claim 1 , wherein reacting the target sequence with both the first unlabeled oligonucleotide and the second unlabeled oligonucleotide in the chamber, to form a 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 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 sequence.
5 . The method of claim 1 , wherein the reaction product is detected through an optically transparent window in the chamber.
6 . The method of claim 1 , wherein the sample 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 at least one of the first unlabeled oligonucleotide and the second unlabeled oligonucleotide comprises a polynucleotide analog.
9 . The method of claim 1 , wherein the first unlabeled oligonucleotide and the second unlabeled oligonucleotide bind to adjacent, contiguous regions of the target sequence.
10 . The method of claim 1 , wherein the more than one optical signal is detected sequentially.
11 . The method of claim 1 , wherein the more than one optical signal is detected simultaneously.
12 . The method of claim 1 , wherein the more than one optical signal comprises at least two distinctive wavelengths.
13 . The method of claim 1 , wherein the more than one optical signal is generated by a fluorescent dyes.
14 . The method of claim 1 , further comprising labeling the reaction product in the chamber, with a detectable label, wherein the detecting comprises detecting an optical signal generated by the label.
15 . The method of claim 14 , wherein the label comprises a fluorescent label.