Method for qualitative and quantitative determination of immunological reactions

• US 4,197,088 A
• Filed: 09/23/1977
• Issued: 04/08/1980
• Est. Priority Date: 09/23/1977
• Status: Expired due to Term

• Current Assignee: Gamma Biologicals, Inc. (Werfen SA)
• Original Assignee: Akro-Medic Engineering Incorporated
• Inventors: Acker, Jesse L., Meserol, Peter M.
• Primary Examiner(s): Marantz, Sidney
• Application Number:
  US05/835,996
• Time in Patent Office: 928 Days
• Field of Search: 23/230 B, 73/64.1, 424/12, 356/39, 250/574
• US Class Current: 436/528
• CPC Class Codes: G01N 2035/00099 Characterised by type of test elements
• CPC Class Codes: G01N 21/03 Cuvette constructions
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/5911 of the scanning type scanning per se G02B
• CPC Class Codes: G01N 21/75 Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated systems in which material is burnt in a flame or plasma G01N21/72, G01N21/73
• CPC Class Codes: G01N 21/82 producing a precipitate or turbidity
• CPC Class Codes: G01N 33/48 Biological material, e.g. blood, urine G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence; Haemocytometers counting blood corpuscules distributed over a surface by scanning the surface G06M11/02
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/5304 Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 35/00029 provided with flat sample substrates, e.g. slides G01N35/028 takes precedence
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/814 Pregnancy
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin

Abstract

A method for qualitative and quantitative determination of agglutination in immunoreactive systems, and for determining the concentration of the immunoreactive components of such system, which method comprises introducing a fluid biological test specimen and a reagent therefor (e.g., latex spheres) into a reaction zone in an image cell and evenly mixing and incubating the mixture thereby forming agglutinated particles. The reaction zone is then transilluminated and the light beams transmitted therethrough are imaged on an image sensor. Dark areas are formed on the surface of the image sensor which are measured, preferably electronically. This procedure is repeated for a reference specimen and the total dark imaged area is compared with the total dark area obtained for the unknown specimen for qualitative determination.

In order to obtain the concentration of the immunoreactive component in the unknown specimen, the foregoing procedure is repeated for at least two specimens with known antibody concentrations, one of which may be a negative control. The concentration of the immunoreactive component of the unknown sample is then determined from the total dark imaged area thereof and the relationship between the total dark imaged areas of the known specimen and their respective concentrations.

Claims

1. A method for qualitative determination of the immunoreactive component of a biological fluid test specimen, which method comprises:

(a) introducing said biological fluid test specimen with a reagent therefor into a reaction zone, said reagent comprising substantially opaque immunologically active colloidal particles,

(b) evenly mixing and incubating said biological fluid test specimen and said reagent in said reaction zone,

(c) transilluminating the contents of said reaction zone with radiant energy and imaging the light beams transmitted therethrough on the surface of an image sensor thereby forming dark areas corresponding to agglutinated particles in said reaction zone.

(d) measuring the total dark areas on the surface of said image sensor,

(e) repeating steps (a) through (d) for a reference sample, and

(f) comparing the total dark imaged areas obtained for said biological fluid test specimen with the total imaged dark areas corresponding to said reference sample.

2. A method as in claim 1 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

3. A method as in claim 1 wherein said incubation is carried out at a temperature of from about 25°

C. to about 50°

C. for about 5 minutes to about 15 minutes.

4. A method as in claim 3 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

5. A method as in claim 1 wherein said reagent is charcoal particles.

6. A method as in claim 5 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

7. A method as in claim 5 wherein said incubation is carried out at a temperature of from about 25°

C. to about 50°

C. for about 5 minutes to about 15 minutes.

8. A method as in claim 7 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

9. A method as in claim 1 wherein said reagent is latex spheres.

10. A method as in claim 9 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

11. A method as in claim 9 wherein said incubation is carried out at a temperature of from about 25°

C. to about 50°

C. for about 5 minutes to about 15 minutes.

12. A method as in claim 11 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

13. A method of determining the concentration of immunoreactive components of biological fluid test specimen, which method comprises:

(a) introducing said biological fluid test specimen with a reagent therefor into a reaction zone, said reagent comprising substantially opaque immunologically active colloidal particles,

(b) evenly mixing and incubating said biological fluid test specimen and said reagent in said reaction zone,

(c) transilluminating the contents of said reaction zone with radiant energy and imaging the light beams transmitted therethrough on the surface of an image sensor thereby forming dark areas corresponding to agglutinated particles in said reaction zone,

(d) measuring the total dark areas on the surface of said image sensor,

(e) repeating steps (a) through (d) for at least two control samples with known but different concentrations of immunoreactive components,

(f) relating the total dark areas of said control specimen with the respective concentrations of said immunoreactive components, and

(g) determining the concentration of said immunoreactive components of said biological fluid test specimen from the relationship obtained in step (f).

14. A method as in claim 13 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

15. A method as in claim 13 wherein said incubation is carried out at a temperature out at a temperature of from about 25°

C. to about 50°

C., for about 5 minutes to about 15 minutes.

16. A method as in claim 15 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

17. A method as in claim 13 wherein said reagent is charcoal.

18. A method as in claim 17 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

19. A method as in claim 17 wherein said incubation is carried out at a temperature of from about 25°

C. to about 50°

C., for about 5 minutes to about 15 minutes.

20. A method as in claim 19 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

21. A method as in claim 13 wherein said reagent is latex spheres.

22. A method as in claim 21 wherein said areas corresponding to the agglutinated particles are measured by electronic means.

23. A method as in claim 21 wherein said incubation is carried out at a temperature of from about 25°

C. to about 50°

C., for about 5 minutes to about 15 minutes.

24. A method as in claim 23 wherein said areas corresponding to the agglutinated particles are measured by electronic means.