Compositions and processes for analyte detection, quantification and amplification
First Claim
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1. A process for detecting or quantifying multiple different nucleic acid species of interest in a library, the process comprising the steps of:
- a) providing;
(i) an array of fixed or immobilized nucleic acids complementary to said nucleic acid species of interest;
(ii) a library of nucleic acid analytes isolated or derived from a biological source, which library of nucleic acid analytes may contain the nucleic acid species of interest sought to be detected or quantified, wherein each of said nucleic acid species of interest comprises at least one inherent universal detection target (UDT); and
(iii) labeled universal detection elements (UDE) which generate a signal directly or indirectly;
b) hybridizing said library (ii) with said array of nucleic acids (i) to form hybrids if said nucleic acid species of interest are present, thereby binding said nucleic acid species of interest comprising said inherent UDTs to said array;
c) contacting said inherent UDTs on said nucleic acid species of interest bound to said array with said labeled UDEs to form a complex between said labeled UDEs and said inherent UDTs, wherein said complex is bound to said array, and wherein the sequence of said labeled UDE consists of a sequence that binds to a 3′
polyA segment, or a consensus sequence when said inherent UDT is a nucleic acid; and
d) detecting or quantifying said multiple different nucleic acid species of interest by detecting or measuring the amount of signal generated from labeled UDEs complexed to inherent UDTs on each nucleic acid species of interest bound to said array.
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Abstract
This invention provides novel compositions and processes for analyte detection, quantification and amplification. Nucleic acid arrays and libraries of analytes are usefully incorporated into such compositions and processes. Universal detection elements, signaling entities and the like are employed to detect and if necessary or desirable, to quantify analytes. Amplification of target analytes are also provided by the compositions and processes of this invention.
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Citations
38 Claims
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1. A process for detecting or quantifying multiple different nucleic acid species of interest in a library, the process comprising the steps of:
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a) providing; (i) an array of fixed or immobilized nucleic acids complementary to said nucleic acid species of interest; (ii) a library of nucleic acid analytes isolated or derived from a biological source, which library of nucleic acid analytes may contain the nucleic acid species of interest sought to be detected or quantified, wherein each of said nucleic acid species of interest comprises at least one inherent universal detection target (UDT); and (iii) labeled universal detection elements (UDE) which generate a signal directly or indirectly; b) hybridizing said library (ii) with said array of nucleic acids (i) to form hybrids if said nucleic acid species of interest are present, thereby binding said nucleic acid species of interest comprising said inherent UDTs to said array; c) contacting said inherent UDTs on said nucleic acid species of interest bound to said array with said labeled UDEs to form a complex between said labeled UDEs and said inherent UDTs, wherein said complex is bound to said array, and wherein the sequence of said labeled UDE consists of a sequence that binds to a 3′
polyA segment, or a consensus sequence when said inherent UDT is a nucleic acid; andd) detecting or quantifying said multiple different nucleic acid species of interest by detecting or measuring the amount of signal generated from labeled UDEs complexed to inherent UDTs on each nucleic acid species of interest bound to said array. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A process for detecting or quantifying multiple different nucleic acid species of interest in a library, the process comprising the steps of:
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a) providing; (i) an array of fixed or immobilized nucleic acids complementary to said nucleic acid species of interest; (ii) a library of nucleic acid analytes isolated or derived from a biological source, which library of nucleic acid analytes may contain the nucleic acid species of interest sought to be detected or quantified, wherein each of said nucleic acid species of interest comprises at least one inherent universal detection target (UDT); and (iii) labeled universal detection elements (UDE) which generate a signal directly or indirectly; b) contacting said inherent UDTs on said nucleic acid species of interest with said labeled UDEs in said library of nucleic acid analytes to form one or more complexes, wherein the sequence of said labeled UDE consists of a sequence that binds to a 3′
polyA segment, or a consensus sequence when said inherent UDT is a nucleic acid;c) hybridizing said library of nucleic acid analytes with said array of nucleic acids (i) to form hybrids if said nucleic acid species of interest are present, wherein said one or more formed complexes are bound to said array; and d) detecting or quantifying said multiple different nucleic acid species of interest by detecting or measuring the amount of signal generated from labeled UDEs complexed to inherent UDTs on each nucleic acid species of interest bound to said array. - View Dependent Claims (21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38)
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Specification