THYROID ANALYTE DETECTION AND MEASUREMENT

• US 20100120169A1
• Filed: 11/09/2009
• Published: 05/13/2010
• Est. Priority Date: 11/12/2008
• Status: Active Grant

• Current Assignee: Hemopet
• Original Assignee: Hemopet
• Inventors: Ongchangco, Ferde S., Dodds, W. Jean
• Granted Patent: US 8,012,769 B2
• Time in Patent Office: Days
• US Class Current: 436/500
• CPC Class Codes: G01N 33/78 Thyroid gland hormones , e.g. T3, T4, TBH, TBG or their receptors

Abstract

A non-radioisotopic method of detecting thyroid analytes comprising detecting T3, Free T3, T4, Free T4 and thyroglobulin autoantibody in a sample of a non-human species. Each one of these analytes in an assay profile includes non-radio isotopic measurement of T3, Free T3, T4, Free T4 and thyroglobulin autoantibody in the sample from the non-human species. A non-radioisotopic method detects T3AA and T4AA thyroid autoantibodies in a sample from a non-human species such as the canine species. Antibodies and autoantibodies are bound, and a precipitated or bound antigen-antigen or antibody—autoantibody complex is formed. The supernatant or surrounding fluid of the bound or precipitated antigen-antigen or antibody-autoantibody complex is then removed. The thyroid activity of the bound complex, precipitate, supernatant or surrounding fluid is measured. The thyroid analyte is at least one of T3, Free T3, T4 or Free T4. A dialysis cell can be used to separate by dialysis the T3AA or T4AA from their respective T3AA or T4AA fractions without radioisotopes. A non-radioisotopic method detects Free T3 and /or Free T4 in a sample of a non-human species.

Claims

1. A non-radioisotopic method of detecting thyroid analytes comprising detecting T3, Free T3, T4, Free T4 and thyroglobulin autoantibody in a sample of a non-human species and wherein each one of these analytes in an assay profile includes non-radio isotopic measurement of T3, Free T3, T4, Free T4 and thyroglobulin autoantibody in the sample from the non-human species.

2. A non-radioisotopic method as claimed in claim 1 and additionally detecting T3AA thyroid autoantibody in a sample from a non-human species which comprises binding antibody or autoantibody, precipitating or binding the antigen-antibody or antigen-autoantibody complex, removing the supernatant or surrounding fluid of the precipitated or bound antigen-antibody or antigen-autoantibody complex;

and measuring the thyroid activity of the bound complex, precipitate, supernatant or surrounding fluid, where the thyroid analyte is T3 or Free T3.

3. A non-radioisotopic method as claimed in claim 1 and additionally detecting T4AA thyroid autoantibody in a sample from a non-human species which comprises binding antibody or autoantibody, precipitating or binding the antigen-antibody or antigen-autoantibody complex, removing the supernatant or surrounding fluid of the precipitated or bound antigen-antibody or antigen-autoantibody complex;

and measuring the thyroid activity of the bound complex, precipitate, supernatant or surrounding fluid, where the thyroid analyte is T4 or Free T4.

19. A non-radioisotopic method as claimed in claim 1 wherein, for a dog species, the r-squared value of T4 for dogs relative to Free T4 is between about 0.75 and about 0.99.

4-18. -18. (canceled)

20. A non-radioisotopic method of detecting thyroid analytes comprising employing a panel for detecting T3, Free T3, T4, and Free T4 in a sample of biological fluid from a non-human species, and wherein each one of these analytes in the panel includes the steps of measuring by a non-radio isotopic measurement of T3, Free T3, T4, and Free T4 in the sample of biological fluid from the non-human species,

21. A method as claimed in claim 20 adding a sample to a luminescence reaction assay for measuring T3, Free T3, T4 and Free T4 in the non-human species, and measuring the amount of luminescence from a reaction of the sample with a reactant in the assay, and thereby determining the amount of T3, Free T3, T4 and Free T4 in the non-human species, including employing a luminescent detector and microparticles as a solid phase for measuring T3, Free T3, T4, Free T4 in the non-human species.

22. A non-radioisotopic method of claim 20 including detecting a thyroid analyte comprising detecting T4 in a sample of biological fluid from a non-human species and wherein the analyte in an assay includes non-radioisotopic measurement of T4, and applying a luminescent detector and microparticles as a solid phase.

23. A non-radioisotopic method as claimed in claim 22 wherein, for dogs, the r-squared value of T4 relative to Free T4 is between about 0.75 and about 0.99.

24. A non-radioisotopic method of claim 20 including applying chemiluminesence, and selectively acridium ester as the label and paramagnetic particles as a solid phase.

25. A non-radioisotopic method of claim 20 including measuring Free T4 by applying a luminescent detector and microparticles as a solid phase.

26. A non-radioisotopic method of claim 20 including measuring Free T4 by chemiluminesence.

27. A non-radioisotopic method of claim 20 including measuring T3 by applying a luminescent detector and microparticles as a solid phase.

28. A non-radioisotopic method of claim 20 including measuring T3 by chemiluminesence.

29. A non-radioisotopic method of claim 20 including measuring Free T3 by applying a luminescent detector and microparticles as a solid phase.

30. A non-radioisotopic method of claim 20 including measuring Free T3 by chemiluminesence.

31. A method as claimed in claim 20 including employing an enzyme-linked immunoassay for measuring the presence of thyroglobulin autoantibody in the non-human species, selected from the group of the canine, feline or equine species.

32. A non-radioisotopic method as claimed in claim 24 wherein, for dogs, the r-squared value of T4 relative to Free T4 is between about 0.75 and about 0.99.

33. A non-radioisotopic method of detecting thyroid analytes comprising employing a panel for detecting T3, Free T3, T4, Free T4 and thyroglobulin autoantibody in a sample of biological fluid from a non-human species, and wherein each one of these analytes in the panel includes the steps of measuring by a non-radio isotopic measurement T3, Free T3, T4, and Free T4 and thyroglobulin autoantibody in the sample of biological fluid from the non-human species, adding a sample to a luminescence reaction assay for measuring T3, Free T3, T4 and Free T4 and thyroglobulin antibody the non-human species, measuring the amount of luminescence from a reaction of the sample with a reactant in the assay, and measuring thyroglobulin autoantibody in the sample by an immunoassay thereby determining the amount of T4, Free T4, T3 and Free T3 and thyroglobulin autoantibody in the non-human species.

34. A non-radioisotopic method of detecting thyroid analytes comprising employing a panel for detecting T3, Free T3, T4 and Free T4 in a sample of biological fluid from a canine species, and wherein each one of these analytes in the panel includes the steps of measuring by a non-radio isotopic measurement T3, Free T3, T4 and Free T4 in the sample of biological fluid from the canine species, adding a sample to a luminescence reaction assay for measuring T3, Free T3, T4 and Free T4 in the canine species, and measuring the amount of luminescence from a reaction of the sample with a reactant in the assay, thereby determining the amount of T3, Free T3, T4 and Free T4 in the canine species, and determining T3AA thyroid autoantibody in the sample by:

a) measuring Free T3 in the sample;

b) treating the sample with a precipitating or binding agent;

c) suspending the precipitate or bound complex, and at least one of;

i. measuring free T3 in suspended precipitate or bound complex, and

ii. subtracting the measurement of i) from a) wherein the result of ii) is directly proportional to the amount of T3AA in the sample;

or

iii. measuring free T3 in supernatant or surrounding fluid of the precipitate or bound complex;

iv. subtracting the measurement of iii) from a) wherein the result of iv) is indirectly proportional to the amount of T3AA in the sample; and

effecting detection of a), c)i or c)iii by employing direct chemiluminesence, and employing a bioluminescent detector and micro particles as a solid phase.