Detection method using both fluorescence and chemiluminescence labels
AssigneeAccess Medical Systems, Ltd.
InventorsHaode Chen, Heng Wu, Hong Tan, Robert F. Zuk
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Abstract
The present invention is directed to an immunoassay method for detecting an analyte in a liquid sample; and provides accuracy and reproducibility for both high concentration and low concentration samples. The method uses both fluorescence marker and chemiluminescence marker, and reads both fluorescence signal and chemiluminescence signal. The analyte concentration is determined based on either a calibration curve for the fluorescence signal, or a calibration curve chemiluminescence signal, using a pre-established analyte concentration value.
Claims20
A method of detecting an analyte in a liquid sample, comprising the steps of:
- (a) dipping a probe tip in a sample solution to bind an analyte, if present, to a first antibody on the probe tip, wherein the probe having a first antibody immobilized on the tip of the probe;
- (b) dipping the probe tip in a reagent solution comprising a second antibody conjugated to a first member of a binding pair to form a first immunocomplex among the analyte, the first antibody, and the second antibody on the probe tip, wherein the first antibody and the second antibody are two different antibodies each against the analyte;
- (c) dipping the probe tip in a wash solution;
- (d) dipping the probe tip in a solution comprising (i) a second member of binding pair labeled with a fluorescent label and (ii) the second member of binding pair labeled with a chemiluminescent label, to form a second immunocomplex having the fluorescent label and a third immunocomplex having the chemiluminescent label on the probe tip;
- (e) dipping the probe tip in a read vessel and measuring the fluorescent signal of the second immunocomplex bound on the probe tip;
- (f) quantitate the analyte concentration based on the fluorescent signal against a first calibration curve;
- (g) dipping the probe tip to a triggering solution to generate a chemiluminescent signal from the third immunocomplex bound on the probe tip;
- (h) quantitate the analyte concentration based on the chemiluminescent signal against a second calibration curve, and
- (i) determining the analyte concentration based on either the fluorescent signal or the chemiluminescent signal.
A method of detecting an analyte in a liquid sample, comprising the steps of:
- (a) mixing a solution comprising a sample, a first antibody conjugated with a hapten, a second antibody conjugated with a first member of a binding pair, wherein the first antibody and the second antibody are two different antibodies each against the analyte;
- (b) dipping a probe tip into the solution of (a) to form a first immunocomplex among the analyte, the first antibody, and the second antibody on the probe tip;
- (c) dipping the probe tip in a wash solution;
- (d) dipping the probe tip in a solution comprising (i) a second member of binding pair labeled with a fluorescent label and (ii) the second member of binding pair labeled with a chemiluminescent label, to form a second immunocomplex having the fluorescent label and a third immunocomplex having the chemiluminescent label on the probe tip;
- (e) dipping the probe tip in a read vessel and measuring the fluorescent signal of the second immunocomplex bound on the probe tip;
- (f) quantitate the analyte concentration based on the fluorescent signal against a first calibration curve;
- (g) dipping the probe tip to a triggering solution to generate a chemiluminescent signal from the third immunocomplex bound on the probe tip;
- (h) quantitate the analyte concentration based on the chemiluminescent signal against a second calibration curve, and
- (i) determining the analyte concentration based on either the fluorescent signal or the chemiluminescent signal.
Dependencies are inferred from the claim text.