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US 6,051,377 AGranted
Multiplex assay for nucleic acids employing transponders
AssigneePharmaseq, Inc.
InventorWlodek Mandecki
Life of this patent
Estimating the term…
Abstract
Disclosed are materials and methods for performing multiplex assays for nucleic acids, in which a transponder is associated with the bead(s) forming the solid phase used in the assay, nucleic acid probes are bound to the surface of the particles, and data concerning the assay is encoded on the transponder. A dedicated read/write device is used to remotely encode or read the data.
Claims19
A method of detecting a target nucleic acid sequence in a sample, comprising the steps of:
- (a) providing a solid phase comprising particles having transponders, the transponders having memory elements, the particles having a nucleic acid probe attached to a surface of the solid phase particles, the probe having a sequence complementary to a target sequence, the transponders having an index number encoded in the memory element;
- (b) contacting the solid phase with a sample to form a sample mixture;
- (c) increasing the amount of the target nucleic acid subjected to analysis by PCR amplification, using at least one oligonucleotide primer which is not immobilized on the solid phase, said PCR amplification comprising at least one cycle of:
- (1) denaturation of nucleic acids in the sample mixture;
- (2) hybridization of nucleic acids in the sample mixture;
- (3) chain extension with DNA polymerase;
- (d) analyzing the solid phase to detect the presence of a label indicative of binding of the target nucleic acid;
- and
- (e) decoding the data encoded on transponders using the dedicated read/write scanner to identify the class of transponders to which analytes are bound.
6 more elements
A method of performing a multiplex solid phase assay for target nucleic acids in a sample, comprising the steps of:
- (a) providing a particulate solid phase, the particles of the solid phase having transponders, the transponders having memory elements, and an oligonucleotide probe attached to a surface of the particle, the oligonucleotide probe complementary to a target sequence;
- p1 (b) the transponders comprising two or more classes of encoded transponders, each class having a different oligonucleotide bound to the surface of the particle, and each class having a different index number encoded on the transponders memory elements;
- (c) contacting the solid phase with a sample to form a sample mixture, the sample mixture containing two more transponders of different classes;
- (d) increasing the amount of the target nucleic acid subjected to analysis by PCR amplification, using at least one oligonucleotide primer which is not immobilized on the solid phase, said PCR amplification comprising at least one cycle of:
- (1) denaturation of nucleic acids in the sample mixture;
- (2) hybridization of nucleic acids in the sample mixture;
- (3) chain extension with DNA polymerase;
- (e) removing unbound sample components from the sample mixture;
- (f) analyzing the solid phase to detect a label indicative of the presence of bound analytes;
- and
- (g) decoding the data encoded on the transponders to identify the class of transponder to which an analyte is bound.
8 more elements
A method of detecting target nucleic acids in a sample, comprising the steps of:
- (a) introducing into the sample at least two populations of solid phase particles, each particle having a transponder and having an oligonucleotide probe attached to its surface, a first population having an oligonucleotide probe that hybridizes to a different target nucleic acid than a second population and the transponders in the first population being encoded with a different identification than the transponders of the second population;
- (b) increasing the amount of the target nucleic acid subjected to analysis by PCR amplification, using at least one oligonucleotide primer which is not immobilized on the solid phase, said PCR amplification comprising at least one cycle of:
- (1) denaturation of nucleic acids in the sample mixture;
- (2) hybridization of nucleic acids in the sample mixture;
- (3) chain extension with DNA polymerase;
- (c) analyzing the particles to detect a label indicating that target nucleic acid has bound to the probe;
- and
- (d) decoding the transponder to identify the probe.
5 more elements
A method of detecting a target nucleic acid sequence in a sample, comprising the steps of:
- (a) providing a solid phase comprising particles having transponders, the transponders having memory elements, the particles having a nucleic acid probe attached to a surface of the solid phase particles, the probe having a sequence complementary to a target sequence, the transponders having an index number encoded in the memory element;
- (b) contacting the solid phase with a sample to form a sample mixture;
- (c) performing at least one cycle of the ligation reaction, said ligation involving at least one oligonucleotide primer which is not immobilized on the solid phase, said ligation cycle comprising the following steps:
- (1) denaturation of nucleic acids in the sample mixture;
- (2) hybridization of nucleic acids in the sample mixture;
- (3) enzymatic reaction with a DNA ligase;
- (d) analyzing the solid phase to detect the presence of a label indicative of binding of the target nucleic acid;
- and
- (e) decoding the data encoded on transponders using the dedicated read/write scanner to identify the class of transponders to which analytes are bound.
6 more elements
A method of performing a multiplex solid phase assay for target nucleic acids in a sample, comprising the steps of:
- (a) providing a particulate solid phase, the particles of the solid phase having transponders, the transponders having memory elements, and an oligonucleotide probe attached to a surface of the particle, the oligonucleotide probe complementary to a target sequence;
- (b) the transponders comprising two or more classes of encoded transponders, each class having a different oligonucleotide bound to the surface of the particle, and each class having a different index number encoded on the transponders memory elements;
- (c) contacting the solid phase with a sample to form a sample mixture, the sample mixture containing two more transponders of different classes;
- (d) performing at least one cycle of the ligation reaction, said ligation involving at least one oligonucleotide primer which is not immobilized on the solid phase, said ligation cycle comprising the following steps:
- (1) denaturation of nucleic acids in the sample mixture;
- (2) hybridization of nucleic acids in the sample mixture;
- (3) enzymatic reaction with a DNA ligase;
- (e) removing unbound sample components from the sample mixture;
- (f) analyzing the solid phase to detect a label indicative of the presence of bound analytes;
- and
- (g) decoding the data encoded on the transponders to identify the class of transponder to which an analyte is bound.
8 more elements
A method of detecting target nucleic acids in a sample, comprising the steps of:
- (a) introducing into the sample at least two populations of solid phase particles, each particle having a transponder and having an oligonucleotide probe attached to its surface, a first population having an oligonucleotide probe that hybridizes to a different target nucleic acid than a second population and the transponders in the first population being encoded with a different identification than the transponders of the second population;
- (b) performing at least one cycle of the ligation reaction, said ligation involving at least one oligonucleotide primer which is not immobilized on the solid phase, said ligation cycle comprising the following steps:
- (1) denaturation of nucleic acids in the sample mixture;
- (2) hybridization of nucleic acids in the sample mixture;
- (3) enzymatic reaction with a DNA ligase;
- (c) analyzing the particles to detect a label indicating that target nucleic acid has bound to the probe;
- and
- (d) decoding the transponder to identify the probe.
5 more elements
A kit for detecting the presence of a nucleic acid in a sample, comprising:
- (a) at least one assay vessel, containing at least one solid phase particle having a transponder, and an oligonucleotide probe bound to a surface of the particle;
- and
- (b) at least one label reagent.
Dependencies are inferred from the claim text.