Detection of enhanced multiplex signals by surface enhanced raman spectroscopy

A method comprising:

• a) providing a target analyte bound to a first specific binding member;
• b) providing a capture reagent bound to a solid substrate, wherein the capture reagent comprises a Raman label;
• c) contacting the target analyte of a) with the capture reagent of b) under conditions suitable for forming a target analyte-capture reagent complex;
• d) contacting, prior to, concurrently with, or subsequent to c) the first specific binding partner with a second specific binding member functionally associated with a seed particle suitable for associating with a SERS substrate, wherein the first specific binding member binds to the second specific binding member and wherein neither the first nor the second specific binding members have Raman labels attached;
• and
• e) contacting the target analyte-capture reagent complex with electromagnetic radiation suitable for detecting a specific property associated with the analyte capture reagent complex by Raman spectroscopy.

A method comprising:

• a) providing a target analyte bound to a first specific binding member;
• b) providing a capture reagent bound to a solid substrate;
• c) contacting the target analyte of a) with the capture reagent of b) under conditions suitable for forming a target analyte-capture reagent complex;
• d) contacting, prior to, concurrently with, or subsequent to c) the first specific binding partner with a second specific binding member functionally associated with a seed particle suitable for associating with a SERS substrate, wherein the first specific binding member binds to the second specific binding member and wherein neither the first nor the second specific binding members have Raman libels attached;
• e) contacting the SERS substrate with a Raman label;
• and
• f) contacting the target analyte-capture reagent complex with electromagnetic radiation suitable for detecting a specific property associated with the analyte-capture reagent complex by Raman spectroscopy.

A test kit comprising:

• a) a container comprising at least one first specific binding member suitable for attachment to a target analyte;
• b) a container comprising at least one second specific binding member that binds to the first specific binding member, wherein the second specific binding member comprises a seed particle suitable for catalyzing the formation of a surface enhanced Raman scattering (SERS) substrate, wherein the SERS substrate can be activated to provide a SERS effect and wherein neither the first nor the second, specific binding members have Raman labels attached;
• c) a composition comprising a solid substrate comprising an array of capture reagents, a plurality of which comprise a Raman label;
• and