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US 5,629,156 AGranted
Multiple capture probe sandwich assays
AssigneeAmoco Corporation
InventorsJyotsna Shah, Walter King, Jing Liu, James Smith, Eugene Serpe, Sonya Popoff
Life of this patent
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Abstract
New methods for performing sandwich assays have been discovered. The assays utilize dual capture probes and eliminate noise resulting from nonspecific binding of detector probe to capture probe.
Claims9
A method for assaying a sample for target nucleic acid comprising the steps of:
- (a) contacting the sample with a first capture probe and detector probe which first capture probe and detector probe each comprise a nucleic acid segment which in the presence of target nucleic acid bind to the target nucleic acid to form a first capture probe:
- target nucleic acid:
- detector probe ternary complex and wherein the detector probe is amplifiable by the enzyme Q.beta. replicase;
- (b) separating the first capture probe:
- target nucleic acid:
- detector probe ternary complex from the sample and introducing the ternary complex to a second medium;
- (c) separating the target nucleic acid:
- detector probe segment of the ternary complex from the first capture probe;
- (d) contacting the second medium with a second capture probe which comprises a nucleic acid which in the presence of target nucleic acid binds to the target nucleic acid to form a second capture probe:
- target nucleic acid;
- detector probe ternary complex;
- (e) separating the second capture probe:
- target nucleic acid:
- detector probe ternary complex from the second medium;
- (f) amplifying such detector probe as is present in the separated second capture probe:
- target nucleic acid:
- detector probe ternary complex with Q.beta. replicase;
- and
- (g) monitoring for the presence or absence of amplification of detector probe.
16 more elements
A method for assaying a sample for target nucleic acid comprising the steps of:
- (a) contacting the sample with a first capture probe and a second capture probe which first and second capture probe each comprise a nucleic acid segment which in the presence of target nucleic acid bind to the target nucleic acid to form a target complex;
- (b) contacting the sample with a second support which binds to the second capture probe;
- (c) separating the second support and bound target complex from the sample and introducing the second support and bound target complex into a second medium;
- (d) separating the target complex from the second support and separating the second support from the second medium;
- (e) contacting the second medium with a first support which binds to the first capture probe;
- (f) separating the first support and bound target complex from the second medium and introducing the first support and bound target complex into a third medium;
- (g) separating the second capture probe:
- target nucleic acid segment of the target complex from the first support and first capture probe and separating the first support and first capture probe from the third medium;
- (h) contacting the third medium with fresh first capture probe and detector probe which is amplifiable by the enzyme Q.beta. replicase and comprises a nucleic acid segment which in the presence of target nucleic acid binds to the target nucleic acid forming a quaternary complex of first capture probe, second capture probe and detector probe bound to target nucleic acid;
- (i) contacting the third medium with fresh first support which binds with the first capture probe;
- (j) separating the first support and bound quaternary complex from the third medium and introducing the first support and bound quaternary complex into a fourth medium;
- (k) separating the first support and bound first capture probe from the second capture probe:
- target nucleic acid:
- detector probe complex and separating the first support and bound first capture probe from the fourth medium;
- (l) amplifying such detector probe as is present in the second capture probe:
- target nucleic acid:
- detector probe complex with Q.beta. replicase;
- and
- (m) monitoring for the presence or absence of amplified detector probe.
16 more elements
Dependencies are inferred from the claim text.