Detection of skatole for meat quality

• US 4,906,563 A
• Filed: 12/28/1987
• Issued: 03/06/1990
• Est. Priority Date: 12/28/1987
• Status: Expired due to Fees

• Current Assignee: Idetek, Inc. (IDEXX Laboratories Incorporated)
• Original Assignee: Idetek, Inc. (IDEXX Laboratories Incorporated)
• Inventors: Singh, Prithipal, Sinha, Ashoke
• Primary Examiner(s): Kepplinger, Esther M.
• Assistant Examiner(s): Scheiner, Toni R.
• Application Number:
  US07/138,305
• Time in Patent Office: 799 Days
• Field of Search: 435/7, 435/18, 435/25, 435/810, 436/518, 436/815, 436/822, 436/809, 436/810
• US Class Current: 435/7.92
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/963 Methods of stopping an enzyme reaction or stabilizing the test materials
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/815 Test for named compound or class of compounds
• CPC Class Codes: Y10S 436/822 Identified hapten

Abstract

A method for detecting boar taint in a pig sample is provided. Anti-skatole antibodies are combined in a reaction buffer with a pig sample to form a reaction mixture. After incubating, the amount of skatole-antibody complex formation is determined. In a particular embodiment, the method utilizes solid substrate-affixed antibodies in a competitive inhibition assay with an enzyme-labeled skatole analogue. Kits facilitating the method are also provided.

Claims

1. A method for detecting boar taint in a pig comprising:

(a) combining in a reaction buffer a sample from said pig and anti-skatole antibodies to form a reaction mixture;

(b) incubating said reaction mixture for a period of time sufficient to form an antibody-skatole complex with any skatole present in said reaction mixture; and

(c) determining the amount of antibody-skatole complex as an indication of boar taint in said pig.

2. The method of claim 1 wherein said sample comprises a member selected from the group consisting of whole blood, serum and plasma.

3. The method of claim 1 wherein said sample comprises backfat extract.

4. The method of claim 1 wherein said antibodies are affixed to a solid support.

5. The method of claim 4 wherein a known amount of labeled skatole or a labeled skatole analogue is added to said reaction mixture together with said sample and said determining comprises detecting the amount of label bound to said solid support.

6. The method of claim 1 wherein said sample is reacted with a known amount of said anti-skatole antibodies for a time sufficient to form an antibody-skatole complex with any skatole present in said sample to form a reaction mixture, said reaction mixture is incubated with solid support affixed skatole or a solid support-affixed skatole analogue and the amount of antibodies either bound to the support or in said reaction mixture is determined.

7. A method for detecting boar taint in a pig comprising:

(a) combining in a reaction buffer a blood sample from said pig and anti-skatole antibodies affixed to a solid support to form a reaction mixture;

(b) incubating said reaction mixture for a period of time sufficient to form an antibody-skatole complex with any skatole present in said reaction mixture;

(c) washing said solid support; and

(d) determining the amount of antibody-skatole complex affixed to said solid support as an indication of boar taint in said sample.

8. A method for detecting boar taint in a pig comprising:

(a) combining in a reaction buffer a labeled skatole analogue, a blood sample from said pig and anti-skatole antibodies affixed to a solid support to form a reaction mixture;

(b) incubating said reaction mixture for a period of time sufficient to form an antibody-skatole complex with any skatole present in said reaction mixture;

(c) washing said solid support; and

(d) determining the amount of said label bound to said solid support in comparison to a sample having a known amount of skatole as an indication of boar taint in said pig.

9. The method of claim 8 wherein said label is an enzyme.

10. The method of claim 9 wherein said enzyme is a hydrolase.

11. The method of claim 9 wherein said enzyme is an oxidoreductase.

12. The method of claim 8 wherein said skatole or skatole analogue is indole-3-butyric acid, 3-carboxymethylindole or 3-methylindole.

13. The method of claim 12, wherein said labeled skatole analogue is an indol-3-butyric acid/alkaline phosphatase conjugate.

14. The method of claim 8, wherein said labeled skatole analogue is a skatole analogue/enzyme conjugate and said determining of step (d) comprises:

(i) adding a substrate for said enzyme to said solid support to form a second reaction mixture;

(ii) incubating said second reaction mixture for a time period sufficient for production of detectable substrate digestion product; and

(iii) measuring the optical density of said second reaction mixture.

15. A method for detecting boar taint in a pig comprising:

(a) combining in a reaction buffer a conjugate of an enzyme with skatole or a skatole analogue, a blood sample from said pig, and anti-skatole antibodies affixed to a solid support to form a first reaction mixture;

(b) incubating said first reaction mixture for a time period sufficient to form an antibody-skatole complex;

(c) washing said solid support with a wash buffer;

(d) incubating a solution containing a substrate for said enzyme with said solid support to form a second reaction mixture; and

(e) determining the optical density of said second reaction mixture in comparison to the optical density of sample having a known amount of skatole as an indication of boar taint in said pig.

16. A method for detecting boar taint in a pig comprising:

(a) combining a blood sample from said pig, indole-3-butyric acid/alkaline phosphatase conjugate in Tris-buffered saline, and anti-skatole antibodies affixed to a microtiter plate to form a first reaction mixture;

(b) incubating said reaction mixture at about 4°

C. for about four hours;

(c) washing said microtiter plate with salinepolyoxylenesorbitan buffer;

(d) incubating a p-nitrophenylphosphate solution with said microtiter plate at about 37°

C. for about one hour to form a second reaction mixture;

(e) altering the pH of said second reaction mixture to stop enzyme activity to form a third reaction mixture; and

(f) determining the optical density of said third reaction mixture in comparison to the optical density of a sample having a known amount of skatole to determine the skatole concentration of said sample,

wherein said pig is identified as having boar said sample has greater than about 0.3 ppm skatole.

17. The method of claim 16 wherein said pig blood sample comprises a member selected from the group consisting of serum and plasma.

18. A kit for determining boar taint in a pig blood sample comprising in separate containers (a) anti-skatole antibodies and (b) labeled skatole or a labeled skatole analogue.

19. The kit of claim 18 wherein said anti-skatole antibodies are affixed to a microtiter plate.

20. The kit of claim 18 wherein said skatole or skatole analogue is labeled with an enzyme.

21. The kit of claim 19 additionally comprising in a separate container skatole.