Nephelometric method for determination of an antigen or antibody content in whole blood

• US 4,889,815 A
• Filed: 10/23/1986
• Issued: 12/26/1989
• Est. Priority Date: 10/25/1985
• Status: Expired due to Fees

• Current Assignee: Alta Diagnostic Machines Limited Immuno Diagnostic Research Laboratory The Medical School University of Birmingham
• Original Assignee: Alta Diagnostic Machines Limited
• Inventors: Deverill, Ian, Bradwell, Arthur R.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Saunders, David A.
• Application Number:
  US07/922,359
• Time in Patent Office: 1,160 Days
• Field of Search: 436/517, 436/536, 436/805, 436/825, 436/909, 436/518
• US Class Current: 436/517
• CPC Class Codes: G01N 21/47 Scattering, i.e. diffuse reflection G01N21/25, G01N21/41 take precedence G01N21/55 takes precedence
• CPC Class Codes: G01N 2201/122 Kinetic analysis; determining reaction rate
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/557 using kinetic measurement, i.e. time rate of progress of an antigen-antibody interaction
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/825 Pretreatment for removal of interfering factors from sample
• CPC Class Codes: Y10S 436/909 Nephelometry

Abstract

A method and apparatus for use in quantifying, in a whole blood sample in which the red cells are lysed, a component which will react with a reagent to form an antigen-antibody complex, the method comprising mixing the sample with the reagent to obtain the complex, exposing the sample to a source of radiation and measuring the intensity of radiation scattered through a given angle by the complex, and the apparatus including a container for receiving the sample which has been treated with the reagent to the component, the container being transparent to radiation having a wavelength falling within a given band width, typically 460-530 nm. A source of radiation within this band width is provided together with a device for detecting the intensity of radiation scattered through a given angle by the sample.

Claims

1. A method of quantifying, in a whole blood sample in which red cells are lysed, an antigen or antibody which will react with a reagent to form an antigen-antibody complex, comprising the steps of:

mixing said sample with said reagent to obtain said complex;

exposing said sample to a source of radiation;

measuring the intensity of radiation scattered through a given angle by said complex, the wavelength of said radiation being selected from the range 460-530 nm; and

relating the measured intensity to the amount of antigen or antibody in said sample.

2. A method as claimed in claim 1, wherein the intensity of said scattered radiation is measured at time intervals, to determine the rate of formation of said antigen-antibody complex.

3. A method as claimed in claim 1, wherein the wavelength of radiation is selected from the range 460-510 nm.

4. A method as claimed in claim 1 in which said radiation is obtained from a xenon flash tube.

5. A method as claimed in claim 1 in which an antigenic protein is quantified.

6. A method as claimed in claim 5 in which said protein is an immunoglobulin.

7. A method of quantifying, in a whole blood sample in which red cells are lysed, a protein which will react with a reagent to form an antigen-antibody complex, comprising the steps of:

mixing said sample with said reagent to obtain said complex;

exposing said sample to a source of radiation;

measuring the intensity of radiation scattered through a given angle by said complex, the wavelength of said radiation being such that the ratio of the intensity of detected scattered radiation to that of the radiation absorbed by haemoglobin from said lysed red cells is maximized; and

relating the measured intensity to the amount of protein in said sample.

8. A method as claimed in claim 7 in which said radiation is obtained from a xenon flash tube.

9. A method as claimed in claim 7 wherein the intensity of the scattered radiation is measured at time intervals, to determine a rate of formation of said antigen-antibody complex.

10. A method as claimed in claim 7 in which said protein is an immunoglobulin.