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Quantitative immunochromatographic assays

  • US 5,753,517 A
  • Filed: 03/29/1996
  • Issued: 05/19/1998
  • Est. Priority Date: 03/29/1996
  • Status: Expired due to Term
First Claim
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1. A method for quantitatively measuring the amount of an analyte of interest in a sample, comprising the steps of:

  • a. obtaining a fluid sample to be assayed for the analyte of interest;

    b. supplying a rapid antigen measurement platform apparatus, the apparatus comprising a membrane strip, the membrane strip comprising an application point, a contact region, a control reaction zone and a detection zone, wherein the contact region is between the application point and the detection zone;

    c. contacting the application point of the membrane strip with the fluid sample;

    d. maintaining the membrane strip under conditions which are sufficient to allow fluid to transport analyte of interest in the fluid sample by capillary action through the strip to the contact region, the contact region having a population of antibody-coated particles imbedded therein, wherein the antibody is an antibody to the analyte of interest, and a population of internal control particles imbedded therein, wherein the internal control particles are coated with an antibody to a control detection reagent;

    e. further maintaining the membrane strip under conditions which are sufficient to allow analyte of interest to bind to the antibody-coated particles, thereby generating contacted antibody-coated particles;

    to allow the fluid in the sample to transport the contacted antibody-coated particles by capillary action through the strip to the detection zone, the detection zone having the analyte of interest immobilized thereon, and to allow the fluid in the sample to transport the internal control particles by capillary action through the strip to the control reaction zone, the control reaction zone having the control detection reagent immobilized thereon;

    to allow the immobilized analyte of interest in the detection zone to bind to contacted antibody-coated particles which are insufficiently coated with the analyte of interest in the fluid sample, and to allow the internal control particles to bind to the control detection reagent; and

    f. detecting the amount of contacted antibody-coated particles bound to the immobilized analyte of interest in the detection zone, and the amount of internal control particles bound to the control detection reagent in the control reaction zone,wherein the amount of analyte of interest in the fluid sample is inversely related to a ratio of the amount of antibody-coated particles bound to the immobilized analyte of interest in the detection zone, to the amount of internal control particles bound to the control detection reagent in the control reaction zone.

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