Patent ID: 8372605

Claim:
A method of detecting an analyte, comprising: (i) combining: (a) an analyte; (b) a first proximity member, comprising a first analyte-specific binding entity that is capable of forming a complex with the analyte and that is conjugated to a first oligonucleotide comprising a first portion and a second portion; (c) a second proximity member, comprising a second analyte-specific binding entity that is capable of forming a complex with the analyte and that is conjugated to a second oligonucleotide comprising a portion that is capable of hybridizing to the first portion of the first oligonucleotide; and (d) a support comprising a capture oligonucleotide that is capable of hybridizing to the second portion of the first oligonucleotide, thereby forming a complex comprising the analyte, the first proximity member, the second proximity member and the capture oligonucleotide on the support, wherein each of the first analyte-specific binding entity and the second analyte-specific binding entity is an antibody or a functional fragment thereof that binds the analyte, wherein the complex further comprises a hybrid comprising the first portion of the first oligonucleotide and the portion of the second oligonucleotide; (ii) the first portion of the first oligonucleotide moiety and the portion of the second oligonucleotide moiety; extending the 3′ terminus of the first oligonucleotide and/or the second oligonucleotide in the complex and producing an amplicon; (iii) amplifying the amplicon and producing an amplification product; and (iv) detecting the analyte by the detection of the amplification product.