Assay verification control for an automated analytical system

• US 5,635,364 A
• Filed: 01/03/1994
• Issued: 06/03/1997
• Est. Priority Date: 03/27/1992
• Status: Expired due to Term

• Current Assignee: Abbott Laboratories Incorporated
• Original Assignee: Abbott Laboratories Incorporated
• Inventors: Raymoure, William J., Mitchell, James E., Clemens, John M., Hills, David B., Tayi, Apparao, Martin, Richard R., Hendrick, Kendall B., Yost, David A., Pennington, Charles D., Kanewske, William J. III, Walker, Donny R., Moore, Larry W., Clark, Frederick L., Smith, B. Jane, Cloonan, Kevin M., Rumbaugh, William D., Spronk, Adrian M., Walker, Edna S., Schrier, Paul R., Merriam, Richard A., Watkins, William E. III, Wohlford, Robert A., Vaught, James A., Winter, Gary E., Lagocki, Peter A., Hance, Robert B., Stanton, Alyn K., McDowell, Douglas D., Clift, Gilbert, Oleksak, Carl M., Schmidt, Linda S., Vickstrom, Richard L.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/176,173
• Time in Patent Office: 1,247 Days
• Field of Search: 422/63, 422/64, 435/7.92, 435/7.93, 435/7.94, 435/962, 435/967, 436/43, 436/172, 436/518, 436/526, 436/533, 436/534, 436/536, 436/538, 436/539, 436/541, 436/805
• US Class Current: 435/7.92
• CPC Class Codes: B01F 31/24 the containers being submitted to a rectilinear movement
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/042 Caps; Plugs
• CPC Class Codes: B01L 2300/043 Hinged closures
• CPC Class Codes: B01L 3/08 Flasks
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: B01L 9/06 Test-tube stands; Test-tube holders
• CPC Class Codes: B29C 45/00 Injection moulding, i.e. forcing the required volume of moulding material through a nozzle into a closed mould; Apparatus therefor injection blow-moulding B29C49/06
• CPC Class Codes: B29K 2995/0032 Birefringent
• CPC Class Codes: B29L 2023/22 Tubes or pipes, i.e. rigid bent tubes not for use as pipe couplings B29L2023/004
• CPC Class Codes: G01N 2001/007 Devices specially adapted for forensic samples, e.g. tamper-proofing, sample tracking
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 2035/00168 Manufacturing or preparing test elements
• CPC Class Codes: G01N 2035/00287 movable lid/cover for sample or reaction tubes
• CPC Class Codes: G01N 2035/00356 Holding samples at elevated temperature (incubation)
• CPC Class Codes: G01N 2035/00386 using fluid heat transfer medium
• CPC Class Codes: G01N 2035/00455 Controlling humidity in analyser
• CPC Class Codes: G01N 2035/00475 Filters
• CPC Class Codes: G01N 2035/00495 : Centrifuges
• CPC Class Codes: G01N 2035/00524 : Mixing by agitating sample carrier
• CPC Class Codes: G01N 2035/00752 : bar codes
• CPC Class Codes: G01N 2035/0093 : random access not determined by physical position
• CPC Class Codes: G01N 2035/0405 : manipulating closing or opening means, e.g. stoppers, screw caps, lids or covers
• CPC Class Codes: G01N 2035/0436 : with pre-packaged reagents, i.e. test-packs
• CPC Class Codes: G01N 2035/0441 : for samples
• CPC Class Codes: G01N 2035/0443 : for reagents
• CPC Class Codes: G01N 2035/0444 : for cuvettes or reaction vessels
• CPC Class Codes: G01N 2035/0446 : Combinations of the above
• CPC Class Codes: G01N 2035/0448 : composed of interchangeable ring elements
• CPC Class Codes: G01N 2035/0453 : Multiple carousels working in parallel
• CPC Class Codes: G01N 2035/0465 : Loading or unloading the conveyor
• CPC Class Codes: G01N 2035/1025 : Fluid level sensing
• CPC Class Codes: G01N 2035/1076 : plurality or independently movable heads
• CPC Class Codes: G01N 2035/1086 : Cylindrical, e.g. variable angle
• 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 21/645 : Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/76 : Chemiluminescence; Bioluminescence
• 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 33/537 : with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/538 : by sorbent column, particles or resin strip , i.e. sorbent materials
• CPC Class Codes: G01N 33/542 : with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/54313 : the carrier being characterised by its particulate form
• CPC Class Codes: G01N 35/0092 : Scheduling
• CPC Class Codes: G01N 35/0099 : comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/025 : having a carousel or turntable for reaction cells or cuvettes
• CPC Class Codes: G01N 35/1004 : Cleaning sample transfer devices
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices
• CPC Class Codes: H01J 49/04 : Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components
• CPC Class Codes: Y10S 435/962 : Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 435/967 : Standards, controls, materials, e.g. validation studies, buffer systems
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis

Abstract

A method for verifying that an assay methodology is properly performed, that assay reagents employed possess the necessary potency for accurately performing such assay methodology, and whether or not test samples or assay reagents have been tampered with or are adulterated, is described. The method is performed by employing an assay verification sample, comprising a positive analyte component and the test sample under analysis, wherein the assay verification sample is analyzed employing the same assay reagents and essentially the same assay methodology employed to analyze the test sample. The method is particularly useful for performing heterogeneous immunoassays on an automated continuous and random access analytical system.

Claims

1. A method for verifying a result in a heterogeneous immunoassay for determining the presence or amount of an analyte present in a test sample, comprising the steps of:

(a) forming a reaction mixture comprising said test sample and a labeled reagent comprising a substance that specifically binds to said analyte labeled with a detectable moiety to form a bound species of said labeled reagent and a free species of said labeled reagent, wherein said bound species of said labeled reagent comprises said analyte and said labeled reagent and said free species of said labeled reagent does not contain said analyte;

(b) dividing said reaction mixture containing said bound species and free species into a first portion and a second portion;

(c) forming an assay verification sample comprising said second portion of said reaction mixture and a positive analyte component, in a known amount, capable of providing a detectable response as an indication of the presence or amount of said positive analyte component present in said assay verification sample;

(d) independently analyzing said first portion of said reaction mixture to provide a first result indicative of the presence or amount of said analyte present in said test sample, said step of analyzing said first portion comprises the steps of (I) separating said free species from said bound species by means of a solid phase material, said solid phase material having a substance that specifically binds to said bound species but not capable of binding said free species, brought into contact with said reaction mixture and (ii) correlating the amount of labeled reagent in said free species or said bound species to the amount or presence of said analyte in said test sample;

(e) independently analyzing said assay verification sample to provide a second result indicative of the presence or amount of said positive analyte component in said assay verification sample, said step of analyzing said assay verification sample comprising the steps of (I) separating said free species from said bound species by means of a solid phase material, said solid phase material having a substance that specifically binds to said bound species but not capable of binding said free species, brought into contact with said assay verification sample and (ii) correlating the amount of labeled reagent in said free species or said bound species to the amount or presence of said analyte present in said assay verification sample; and

(f) determining if said second result is as expected, given knowledge that said assay verification sample contains said positive analyte component or said positive analyte component in said known amount, to verify said first result.

2. The method of claim 1, wherein the steps of forming said reaction mixture and said assay verification sample are performed sequentially.

3. The method of claim 1, wherein said positive analyte component is said analyte or an analog of said analyte.

4. The method of claim 1, wherein said solid phase material is selected from the group consisting of beads, particles, and microparticles.

5. A method for verifying a result in a heterogeneous immunoassay for determining the presence or amount of an analyte present in a test sample, wherein said heterogeneous assay is a competitive immunoassay, comprising the steps of:

(a) forming a mixture comprising said test sample and a labeled reagent comprising said analyte or an analog of said analyte labeled with a detectable moiety;

(b) dividing said mixture into a first portion and a second portion;

(c) forming an assay verification sample comprising said second portion of said reaction mixture and a positive analyte component, in a known amount, capable of providing a detectable response as an indication of the presence or amount of said positive analyte component present in said assay verification sample;

(d) independently analyzing said first portion of said reaction mixture to provide a first result indicative of the presence or amount of said analyte present in said test sample, said step of analyzing said first portion comprising the steps of (I) contacting said mixture with a solid phase material, said solid phase material having a substance that specifically binds to said analyte or said labeled reagent to form a bound species of said labeled reagent and a free species of said labeled reagent and (ii) correlating the amount of said bound species of said labeled reagent or said free species of said labeled reagent to the amount or presence of said analyte present in said test sample;

(e) independently analyzing said assay verification sample to provide a second result indicative of the presence or amount of said positive analyte component in said assay verification sample, said step of analyzing said assay verification sample comprising the steps of (I) contacting said mixture with a solid phase material, said solid phase material having a substance that specifically binds to said analyte or said labeled reagent to form a bound species of said labeled reagent and a free species of said labeled reagent and (ii) correlating the amount of said bound species of said labeled reagent or said free species of said labeled reagent to the amount or presence of said analyte present in said assay verification sample; and

(f) determining if said second result is as expected, given knowledge that said assay verification sample contains said positive analyte component or said positive analyte component in said known amount, to verify said first result.

6. A method for verifying a result obtained in an analytical procedure for determining the presence or amount of an analyte in a test sample, comprising the steps of:

(a) forming a reaction mixture comprising said test sample and at least one appropriate assay reagent suitable for analysis of said test sample for said analyte;

(b) independently analyzing said reaction mixture to provide a first result indicative of the presence or amount of said analyte in said test sample;

(c) forming an assay verification sample comprising said independently analyzed reaction mixture of step (b) and a positive analyte component capable of providing a detectable response as an indication of the presence or amount of said positive analyte component present in said assay verification sample;

(d) independently analyzing said assay verification sample to provide a second result indicative of the presence or amount of said positive analyte component in said assay verification sample; and

(e) determining if said second result is as expected, given knowledge that said assay verification sample contains said positive analyte component or said positive analyte component in said known amount, to verify said first result.

7. The method of claim 6, wherein said second result indicates said first result could be inaccurate if said second result is not as expected given knowledge that said assay verification sample contains said positive analyte component.

8. The method of claim 6, wherein said second result indicates said first result is accurate if said second result is as expected given knowledge that said assay verification sample contains said positive analyte component.

9. The method of claim 6, wherein the steps of forming said reaction mixture and said assay verification sample are performed simultaneously.

10. The method of claim 6, wherein the steps of forming said reaction mixture and said assay verification sample are performed sequentially.

11. The method of claim 6, wherein said positive analyte component is selected from the group consisting of said analyte and analogs of said analyte.

12. The method of claim 6 wherein said analytical procedure is an immunoassay.

13. The method of claim 12, wherein said immunoassay is a homogeneous immunoassay.

14. The method of claim 12, wherein said immunoassay is a heterogeneous immunoassay.

15. A method for verifying a result in a heterogeneous immunoassay for determining the presence or amount of an analyte present in a test sample, comprising the steps of:

(a) forming a reaction mixture comprising said test sample and a labeled reagent comprising a substance that specifically binds to said analyte labeled with a detectable moiety to form a bound species of said labeled reagent and a free species of said labeled reagent, wherein said bound species of said labeled reagent comprises said analyte and said labeled reagent and said free species of said labeled reagent does not contain said analyte;

(b) dividing said reaction mixture containing said bound species and said free species into a first portion and a second portion;

(c) forming an assay verification sample comprising said second portion of said reaction mixture and a positive analyte, in a known amount, capable of providing a detectable response as an indication of the presence or amount of said positive analyte component present in said assay verification sample;

(d) independently analyzing said first portion of said reaction mixture to provide a first result indicative of the presence or amount of said analyte present in said test sample, said step of analyzing said first portion comprising the steps of (I) separating said free species from said bound species by means of a solid phase material, said solid phase material having a substance that specifically binds to said bound species but not capable of binding said free species, brought into contact with said reaction mixture and (ii) correlating the amount of labeled reagent in said free species or said bound species to the amount or presence of said analyte present in said test sample;

(e) adding assay verification sample to said independently analyzed first portion of said reaction mixture to provide a second result indicative of the presence or amount of said positive analyte component in said assay verification sample, said step of analyzing said assay verification sample comprising the steps of (I) separating said free species from said bound species by means of a solid phase material, said solid phase material having a substance that specifically binds to said bound species but not capable of binding said free species, brought into contact with said assay verification sample and (ii) correlating the amount of labeled reagent in said free species or said bound species to the amount or presence of said analyte present in said assay verification sample; and

(f) determining if said second result is as expected, given knowledge that said assay verification sample contains said positive analyte component or said positive analyte component in said known amount, to verify said first result.

16. The method of claim 15, wherein the steps of forming said reaction mixture and said assay verification sample are performed sequentially.

17. The method of claim 15, wherein said positive analyte component is said analyte or an analog of said analyte.

18. The method of claim 15, wherein said solid phase material is selected from the group consisting of beads, particles, and microparticles.

19. A method for verifying a result in a heterogeneous immunoassay for determining the presence or amount of an analyte present in a sample, wherein said heterogeneous immunoassay is a competitive heterogeneous immunoassay, comprising the steps of:

(a) forming a mixture comprising said test sample and a labeled reagent comprising said analyte or an analog of said analyte labeled with a detectable moiety;

(b) dividing said mixture into a first portion and a second portion;

(c) forming an assay verification sample comprising said second portion of said reaction mixture and a positive analyte component, in a known amount, capable of providing a detectable response as an indication of the presence or amount of said positive analyte component present in said assay verification sample;

(d) independently analyzing said first portion of said reaction mixture to provide a first result indicative of the presence or amount of said analyte present in said test sample, said step of analyzing said first portion comprising the steps of (I) contacting said mixture with a solid phase material, said solid phase material having a substance that specifically binds to said analyte or said labeled reagent to form a bound species of said labeled reagent and a free species of said labeled reagent and (ii) correlating the amount of said bound species of said labeled reagent or said free species of said labeled reagent to the amount or presence of said analyte present in said test sample;

(e) independently analyzing said assay verification sample to provide a second result indicative of the presence or amount of said positive analyte component in said assay verification sample, said step of analyzing said assay verification sample comprising the steps of (I) contacting said mixture with a solid phase material, said solid phase material having a substance that specifically binds to said analyte or said labeled reagent to form a bound species of said labeled reagent and a free species of said labeled reagent and (ii) correlating the amount of said bound species of said labeled reagent or said free species of said labeled reagent to the amount or presence of said analyte present in said assay verification sample; and

(f) determining if said second result is as expected, given knowledge that said assay verification sample contains said positive analyte component or said positive analyte component in said known amount, to verify said first result.