Multiplexed detection of analytes
First Claim
Patent Images
1. A method of detecting at least first and second target molecules in a sample comprising:
- a) contacting said first and second target molecule with a composition comprising;
i) an amplification enzyme; and
ii) first and second target probes, said first and second target probes comprising;
(a) a first and a second bioactive agent, respectively, wherein said first and second bioactive agents specifically bind to said first and second target molecules, respectively;
(b) a first and a second adapter sequence, respectively, wherein said first adapter sequence identifies said first target molecule and said second adapter sequence identifies said second target molecule; and
(c) at least a first and a second upstream universal priming sequences;
b) amplifying said first and second adapter sequences using said first and second universal priming sequences, wherein no ligation is performed, to form first and second amplicons, respectively;
c) detecting said first and second amplicons, respectively, to indicate the presence of said first and second target molecules in said sample.
1 Assignment
0 Petitions
Accused Products
Abstract
The present invention is directed to sensitive and accurate multiplexed assays for target analyte detection.
424 Citations
34 Claims
-
1. A method of detecting at least first and second target molecules in a sample comprising:
-
a) contacting said first and second target molecule with a composition comprising;
i) an amplification enzyme; and
ii) first and second target probes, said first and second target probes comprising;
(a) a first and a second bioactive agent, respectively, wherein said first and second bioactive agents specifically bind to said first and second target molecules, respectively;
(b) a first and a second adapter sequence, respectively, wherein said first adapter sequence identifies said first target molecule and said second adapter sequence identifies said second target molecule; and
(c) at least a first and a second upstream universal priming sequences;
b) amplifying said first and second adapter sequences using said first and second universal priming sequences, wherein no ligation is performed, to form first and second amplicons, respectively;
c) detecting said first and second amplicons, respectively, to indicate the presence of said first and second target molecules in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
-
-
29. A method for multiplex detection of a plurality of target molecules in a sample, said method comprising;
-
a) contacting said plurality of target molecules with a composition comprising a plurality of target probes, each comprising;
i) a bioactive agent, wherein each bioactive agent binds to a unique target molecule;
ii) an adapter sequence that identifies said unique target molecule that binds the bioactive agent; and
iii) at least one upstream universal priming sequence;
b) removing unbound target probes, leaving bound target probes;
c) contacting said bound target probes with an amplification enzyme;
d) amplifying said adapter sequences of said bound target probes using said at least one universal priming sequence, wherein no ligation is performed, to form a plurality of amplicons;
e) detecting said plurality of amplicons, to indicate the presence of said target molecules in said sample. - View Dependent Claims (30, 31, 32, 33, 34)
-
Specification