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 detecting or quantitating at least one polynucleotide sequence in a liquid sample comprising:
providing a device comprising a substrate defining a sample-distribution network having (i) a sample inlet, (ii) two or more detection chambers, and (iii) a channel in fluid communication with each of the detection chambers and the sample inlet, wherein at least one of the detection chambers contains pre-loaded and dried down first contents comprising a first sequence-specific polynucleotide binding polymer for detecting or quantitating a polynucleotide sequence, and at least a different one of the detection chambers contains pre-loaded and dried down second contents comprising a second sequence-specific polynucleotide binding polymer for detecting or quantitating a polynucleotide sequence, that differs from the first sequence-specific polynucleotide binding polymer;
loading a liquid sample comprising at least one polynucleotide sequence into the two or more detection chambers until the two or more detection chambers are loaded with the liquid sample such that the loading is stopped;
after the two or more detection chambers are loaded with liquid sample and the loading is stopped, isolating the two or more detection chambers from one another and isolating the first contents from the second contents such that the liquid sample is prevented from flowing from one of the detection chambers to another of the detection chambers;
reacting the loaded sample with at least one of the sequence-specific polynucleotide binding polymers under conditions effective to produce a detectable signal in at least one of the detection chambers; and
measuring the signal produced in the detection chambers to detect or quantitate the at least one polynucleotide sequence in the sample.
2. The method of claim 1 , wherein the channel comprises a single channel.
3. The method of claim 1 , wherein the channel comprises a first channel with which a first group of detection chambers is in fluid communication, and a second channel with which a second group of detection chambers is in fluid communication.
4. The method of claim 1 , wherein the channel comprises a plurality of branch channels, one branch channel in fluid communication for with each detection chamber.
5. The method of claim 1 , wherein at least one of the first and second sequence-specific polynucleotide binding polymers comprises an oligonucleotide primer pair suitable for amplifying, by polymerase chain reaction, a specific polynucleotide sequence that is flanked by sequences complementary to the primer pair.
6. The method of claim 1 , wherein the isolating comprises clearing the channel of liquid sample after the two or more detection chambers are loaded with the liquid sample.
7. The method of claim 1 , wherein the isolating comprises filling the channel with a fluid.
8. The method of claim 1 , wherein the reacting occurs in the detection chambers after the isolating, and wherein the detectable signal is produced in a plurality of the detection chambers.
9. The method of claim 1 , wherein the loading comprises dividing the liquid sample into sample portions each contained within respective detection chambers, and wherein the reacting occurs in one of the sample chambers for a respective sample portion, such that the detectable signal is produced in a plurality of the detection chambers.