Nanoparticle probs with Raman spectrocopic fingerprints for analyte detection

A method for detecting for the presence or absence of one or more target analytes in a sample, the target analytes having at least two binding sites, said method comprising:

• providing a substrate having bound thereto one or more types of a first specific binding member for immobilizing the target analyte onto said substrate;
• providing one or more types of particles having bound thereto (a) one or more Raman labels;
• and (b) a second specific binding member for binding to a specific target analyte, wherein (i) the Raman active label bound to each type of particle is different and serves as an identifier for a specific target analyte;
• (ii) the second specific binding member bound to each type of particle is different and is targeted to a specific target analyte;
• and (iii) the Raman label comprises at least one Raman label.
• contacting the particles, the sample and the substrate under conditions effective for specific binding interactions between the target analyte and first and second specific binding member so as to form a test substrate having particles complexed thereto in the presence of one or more target analytes in the sample;
• contacting the test substrate with a staining material to produce a detection substrate having a surface capable of causing surface-enhanced Raman scattering (SERS);
• and
• determining for the presence of said particle complexes on said detection substrate as an indication of the presence of one or more target analytes in the sample by obtaining and analyzing a SERS spectrum.

A method for detecting for the presence or absence of one or more target nucleic acids in a sample, the sequence of the nucleic acid having at least two portions, said method comprising:

• providing a substrate having oligonucleotides bound thereto, the oligonucleotides bound to the substrate having a sequence that is complementary to a first portion of the nucleic acid;
• providing one or more types of particles comprising oligonucleotides bound thereto and a Raman active label bound to a portion of the oligonucleotides, wherein (i) at least some of the oligonucleotides attached to each type of particle have a sequence that is complementary to a second portion of the sequence of a specific target nucleic acid;
• and (ii) the Raman label bound to each type of particles is different and serves as an identifier for a specific target nucleic acid, said Raman label comprising at least one Raman label.
• contacting the particles, the substrate, and the sample under conditions effective for hybridization of the oligonucleotides bound to the substrate with the first portion of the nucleic acid and for hybridization of the oligonucleotides attached to the particle with the second portion of the nucleic acid so as to form a test substrate having one or more particle complexes bound thereto when one or more target nucleic acids are present in said sample;
• contacting the test substrate with a staining material to produce a detection substrate having a surface capable of causing surface-enhanced Raman scattering (SERS);
• and
• determining for the presence of said particle complexes on said detection substrate as an indication of the presence of one or more target nucleic acids in the sample by obtaining and analyzing a SERS spectrum.

A method for detecting for the presence or absence of a target nucleic acid in a sample, the sequence of the nucleic acid having at least two portions, said method comprising:

• providing a substrate having oligonucleotides bound thereto, the oligonucleotides bound to the substrate having a sequence that is complementary to a first portion of the nucleic acid;
• providing a particle comprising oligonucleotides bound thereto and a Raman label bound to a portion of the oligonucleotides, wherein (i) at least some of the oligonucleotides attached to the particle have a sequence that is complementary to a second portion of the nucleic acid;
• and (ii) the Raman label bound to particles serves as an identifier for the target nucleic acid, said Raman label comprising at least one Raman label providing a detectable or measurable Raman scattering signal when illuminated by radiation capable of inducing a Raman scattering;
• contacting the particles, the substrate, and the sample under conditions effective for hybridization of the oligonucleotides bound to the substrate with the first portion of the nucleic acid and for hybridization of the oligonucleotides attached to the particle with the second portion of the nucleic acid so as to form a test substrate having a particle complex bound thereto when said target nucleic acid is present in said sample;
• contacting the test substrate with a staining material to produce a detection substrate having a surface capable of causing surface-enhanced Raman scattering (SERS);
• and
• determining for the presence of said particle complex on said detection substrate as an indication of the presence of the target nucleic acid in the sample by obtaining and analyzing a SERS spectrum.

A method for screening one or more molecules to determine whether the molecule is a ligand to one or more specific receptors, the molecules are present in a sample, said method comprising:

• providing a substrate having bound thereto one or more specific receptors;
• providing conjugates comprising particles, oligonucleotides bound to the particles, a Raman active label bound to a portion of the oligonucleotides, and the molecule from said sample bound to a portion of the oligonucleotides, wherein said Raman active label comprising at least one Raman active molecule providing a detectable or measurable Raman scattering signal when illuminated by radiation capable of inducing a Raman scattering;
• contacting the particles, sample and substrate under conditions effective for specific binding interactions between the molecule bound to the particles with the specific receptor bound to the substrate so as to form a test substrate having particles complexed thereto when the molecule is a ligand to a specific receptor;
• contacting the test substrate with a staining material to produce a detection substrate having a surface capable of causing surface-enhanced Raman scattering (SERS);
• and
• determining for the presence of said particle complexes on said detection substrate as a confirmation of a ligand to a specific receptor by obtaining and analyzing a SERS spectrum.