Method for detecting the presence of an analyte in a sample

• US 7,097,983 B2
• Filed: 09/24/2001
• Issued: 08/29/2006
• Est. Priority Date: 07/16/1997
• Status: Expired due to Term

• Current Assignee: Charm Sciences, Inc.
• Original Assignee: Charm Sciences, Inc.
• Inventors: Markovsky, Robert J., Donahue, Paul R., Scheemaker, Joan L., Boyer, Cheryl A., Trivedi, Shefali B., Glickman, Yael Agi, Charm, Stanley E., Saul, Steven J., Skiffington, Richard T., Zomer, Eliezer
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): CHEU, CHANGHWA J
• Application Number:
  US09/961,998
• Publication Number:
  US 20030207442A1
• Time in Patent Office: 1,800 Days
• Field of Search: 435/7.1, 435/7.92, 435/970, 435/805, 436/501, 436/514, 436/530, 436/535, 436/536, 436/524, 436/525, 436/8, 436/15, 422/56, 422/60, 422/61
• US Class Current: 435/7.1
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A23K 10/30 from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms obtained by microbiological or biochemical processes, e.g. using yeasts or enzymes, A23K10/10
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: C11B 3/001 by a combination of two or more of the means hereafter
• CPC Class Codes: C11B 7/00 Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
• CPC Class Codes: C11C 1/04 by hydrolysis
• CPC Class Codes: C11C 3/10 Ester interchange
• CPC Class Codes: C11C 3/123 using catalysts based principally on nickel or derivates
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/8222 Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
• CPC Class Codes: C12N 15/8234 Seed-specific, e.g. embryo, endosperm
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/0083 Miscellaneous (1.14.99)
• CPC Class Codes: G01N 2201/0221 Portable; cableless; compact; hand-held
• CPC Class Codes: G01N 33/537 with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/105831 : Protein or peptide standard or control [e.g., hemoglobin, etc.]

Abstract

The invention features a method of determining whether one or more members of an analyte family are present in a sample. The method makes use of a test zone binder, e.g., a receptor that can bind one or a plurality of analytes within an analyte family, the analytes family defined by similar structural binding sites. Members of an analyte family can have different detection level requirements and, therefore, additional analyte binders can be employed to adjust test sensitivity for a subset of the analytes in the analyte family individually, so that each analyte can be detected only if it is present in the sample above a predetermined threshold.

Claims

1. A method for determining whether one or more members of an analyte family is present in a test sample at or above a predetermined threshold level, wherein the predetermined threshold level for one member of said analyte family differs from the predetermined threshold level for at least one other member of said analyte family, said method comprising the steps of:

(a) allowing said sample to contact a receptor, wherein said receptor is characterized by an ability to bind to at least two members of said analyte family, so as to allow said receptor to bind said at least two members of said analyte family in said sample, if present, to form an analyte-receptor complex with said present member, whereby said method has an initial analyte-specific test sensitivity level for each of said at least two members respectively;

(b) prior to, concurrently with, or subsequent to step (a), allowing said sample to contact an amount of one or more analyte-specific adjustment binders, each adjustment binder having binding specificity for at least one selected member of said analyte family but not all of said at least two members of the analyte family, said analyte-specific adjustment binder competing with said receptor for binding to said at least one selected member, the amount of each of said one or more analyte-specific adjustment binders chosen to make an amount of said at least one selected member unavailable for binding to said receptor, said amount of unavailable selected member being sufficient to adjust said selected member'"'"'s respective sensitivity level to said selected member'"'"'s predetermined threshold level;

(c) subsequent to steps (a) and (b), allowing said sample to come into contact with a receptor binder on a solid support, said receptor binder characterized in that said receptor binder binds to the receptor but does not bind to the analyte-receptor complex;

(d) prior to, concurrently with, or subsequent to steps (a)–

(c), allowing said receptor to be tagged with a detectable label; and

(e) detecting receptor bound to said receptor binder on said solid support as an inverse indication of the presence of one or more of said members of said analyte family in said sample at or above said predetermined threshold level.

2. The method of claim 1, further comprising the steps of:

(f) subsequent to steps (a) and (b), allowing said sample to come into contact with a control binder, said control binder characterized in that it binds both to said receptor and to said analyte-receptor complex;

(g) detecting receptor and analyte-receptor complex bound to said control binder; and

(h) comparing binding to said control binder with binding to said receptor binder, whereby said one or more members of said analyte family is present in said sample at or above said threshold level when said binding to said control binder exceeds binding to said receptor binder.

3. The method of claim 1, wherein said receptor is an enzyme.

4. The method of claim 1, wherein said receptor is an antibody.

5. The method of claim 1, wherein said receptor is isolated from B. stearothermophilus.

6. The method of claim 1, further comprising a second receptor, said second receptor having binding affinity for at least one additional analyte in said sample.

7. The method of claim 1, further comprising one or more additional receptors which bind analytes within the same analyte family to which the first receptor is able to bind, said additional receptor having a different sensitivity for one or more analytes within said analyte family as compared to said first receptor.

8. The method of claim 7, wherein said additional receptor binds to cloxacillin.

9. The method of claim 1, wherein the receptor binder comprises an analyte conjugate.

10. The method of claim 1, wherein said receptor binder comprises ceforanide linked to a protein.

11. The method of claim 2, wherein said control binder is a rabbit anti-receptor antibody.

12. The method of claim 1, wherein said sample is allowed to contact said receptor at a temperature of 30°

C. or greater.

13. The method of claim 1, wherein said adjustment binder is selected from the group consisting of an enzyme, a receptor, and an antibody.

14. The method of claim 1, wherein the adjustment binder is specific for an analyte selected from the group consisting of ceftiofur, cephapirin, amoxicillin, ampicillin and penicillin.

15. The method of claim 1, wherein the adjustment binder is specific for cloxacillin.

16. The method of claim 1, wherein the sample is allowed to contact the adjustment binder prior to contact with the receptor.

17. The method of claim 1, wherein a result that said analyte is present in said sample at or above a threshold level is a positive result, and a positive does not differentiate one particular analyte from another.

18. The method of claim 1, wherein the receptor is bound to detectable microparticles.

19. The method of claim 1, wherein the receptor comprises protein-coated, gold microsphere beads.

20. The method of claim 1, wherein the method detects cephapirin at a sensitivity of about 15 to 20 ppb.

21. The method of claim 1, wherein the method detects ceftiofur at a sensitivity of about 40 to 50 ppb.

22. The method of claim 1, wherein said analyte is a toxin.

23. The method of claim 1, wherein said analyte is a beta lactam.

24. The method of claim 1, wherein said analyte is a tetracycline.

25. The method of claim 1, wherein said analyte is a sulfonamide.

26. The method of claim 1, wherein said analyte is a macrolide.

27. The method of claim 1, wherein said analyte is an aminoglycoside.

28. The method of claim 1, wherein said analyte is a quinolone.

29. The method of claim 1, wherein said analyte is a pesticide.

30. The method of claim 1, wherein said analyte is a microorganism.

31. The method of claim 1, wherein said method further comprises the steps of:

(a) providing a test device, said test device comprising;

(i) a support strip;

(ii) a sample-absorbing matrix attached to said support strip, said sample-absorbing matrix having material for absorbing an amount of the sample;

(iii) a mobile-phase support for holding a mobile-phase composition, said mobile-phase support and mobile-phase composition being in fluid communication with said sample absorbing matrix, wherein said mobile-phase composition comprises said receptor and one or more of said analyte-specific adjustment binders;

(iv) a stationary-phase membrane in contact with the mobile-phase support and having a first end and a second end, wherein said membrane allows lateral capillary flow of the sample from the first end to the second end; and

(v) at least one test zone on the membrane having said receptor binder bound to said membrane; and

(b) applying said sample to said sample-absorbing matrix.

32. The method of claim 31, wherein said test device further comprises a control zone on the membrane, said control zone comprising a control binder characterized in that it binds both to said receptor and to said analyte-receptor complex, said method further comprising the step of detecting receptor and analyte-receptor complex bound to said control binder at said control zone.

33. The method of claim 31, wherein the sample-absorbing matrix includes a dry, compressed, cellulosic-membrane material.

34. The method of claim 31, wherein said test device further includes a disposal zone at the second end.

35. The method of claim 31, wherein said test device further includes an elongate blister housing enclosing said support strip, sample-absorbing matrix, mobile-phase support, mobile-phase composition, stationary-phase membrane, test zone and control zone, said elongate blister housing defining an elongated strip cavity having a first end and a second end.

36. The method of claim 35, wherein the elongated blister housing comprises a transparent blister housing secured to a tape strip having a peelable end tab.

37. The method of claim 35, wherein said elongate blister housing includes a transparent, top-cover section to allow observation of test results on the test device.

38. The method of claim 35, wherein said elongate blister housing is characterized by an expansion cavity housing at the one end and extending outwardly from a top cover.

39. The method of claim 38, wherein the expansion cavity includes a top cover which has one or more apertures therein to increase the penetration efficiency of the liquid sample into the sample-absorbing matrix.

40. The method of claim 35, wherein the elongate blister housing is formed of a transparent plastic material.

41. The method of claim 35, wherein the first end comprises a means for sealing an expansion cavity.

42. The method of claim 35, wherein the elongated blister housing includes an expansion cavity at the one end of the housing and opposite to the sample-absorbing matrix, to permit selected expansion of the sample-absorbing matrix to fill the expansion cavity.

43. The method of claim 32, wherein said detecting comprises observing a signal, said method further comprising the step of comparing the detectable signal of the control zone to the detectable signal of the test zone, the test device arranged and constructed to provide a positive or negative result when a member of the analyte family is present at or above a threshold level, said member having a threshold level which is different from the threshold level of other members of the analyte family, said positive result indicated by a more intense signal in the control zone as compared to the test zone and said negative result indicated by a less intense signal in the control zone as compared to the test zone.

44. The method of claim 2, further comprising the step of using a reader for said comparing.

45. The method of claim 31, wherein said mobile-phase composition further comprises one or more additional receptors which bind analytes within the same analyte family to which the first receptor is able to bind, said additional receptor having a different sensitivity for one or more analytes within said analyte family as compared to said first receptor.

46. The method of claim 31, wherein said mobile-phase composition further comprises a second receptor, said second receptor having binding affinity for at least one additional analyte in said sample.

47. The method of claim 46, wherein said device comprises at least two test zones, wherein a first test zone of said test zones comprises a first receptor binder characterized in that said first receptor binder binds to said first receptor but does not bind to the analyte receptor-complex, and wherein a second test zone of said test zones comprises a second receptor binder characterized in that said second receptor binder binds to the second receptor but does not bind to the analyte-receptor complex.