초록 ▼

A waveguide binding assay method involves detecting the scattering of light directed into the waveguide, the scattering being the result of scattering labels specifically bound to the waveguide within the penetration depth of an evanescent wave. The waveguide may be transparent plastic or glass and

A waveguide binding assay method involves detecting the scattering of light directed into the waveguide, the scattering being the result of scattering labels specifically bound to the waveguide within the penetration depth of an evanescent wave. The waveguide may be transparent plastic or glass and the binding is typically by oligonucleotide hybridization or immunological capture. Light scattering labels include colloidal metals or non-metals, including gold, selenium and latex. A light absorbing member consisting of dye or concentrated particles may also be employed to enhance signal. Real-time binding and dissociation can be monitored visually or by video imaging, such as with a CCD camera and frame grabber software. Hybridization mismatches of as few as one base can be distinguished by real-time melting curves.

대표청구항 ▼

A method for detecting the presence or amount of one or more specific binding analytes in a fluid sample, the method comprising: (a) providing a waveguide device, the waveguide device comprising (i) a transparent element having a refractive index greater than that of the fluid sample; (ii) a light r

A method for detecting the presence or amount of one or more specific binding analytes in a fluid sample, the method comprising: (a) providing a waveguide device, the waveguide device comprising (i) a transparent element having a refractive index greater than that of the fluid sample; (ii) a light receiving edge; and (iii) a reactive surface comprising a first specific binding member of at least one cognate binding pair immobilized at a plurality of sites on the surface of the element, other non-situs portions of the reactive surface having no specific binding member immobilized thereon; wherein said first specific binding member, through intermediate cognate binding pairs if desired, specifically binds at least one analyte; (b) contacting the reactive surface with a sample suspected to contain said one or more analytes and with a light scattering label attached to a specific binding member of a second cognate binding pair which, through intermediate cognate binding pairs if desired, specifically binds said one or more analytes, in the case of a sandwich assay, or the immobilized first specific binding member of said first cognate binding pair, in the case of a competitive assay; thereby forming light scattering label complexes attached to the plurality of sites in proportion to the amount of analyte in the sample; (c) illuminating the light receiving edge of the waveguide with light effective to create total internal reflection within the waveguide, thereby simultaneously illuminating the entire reactive surface; (d) collecting visually detectable light scattered by said light scattering label; (e) comparing the degree of light scattering at each situs with either (i) the degree of light scattering at a non-situs portion, or (ii) the degree of light scattering at another situs, or both, whereby light scattering at each situs correlates to the presence or amount of the analyte for which the immobilized specific binding member at that situs is specific.