Device for detection of vitamin D metabolites

1. A test device for detecting one or more vitamin D molecules comprising:

a housing, containing therein:

(a) a sample application pad configured to absorb a biological sample and transport said biological sample to a conjugate pad;

(b) said conjugate pad, wherein said conjugate pad comprises a vitamin D binding agent that specifically binds to one or more vitamin D molecules; and

(c) a detection zone comprising a first region having immobilized thereon a detection agent that specifically binds an epitope that is generated by complexing said vitamin D binding agent with said one or more vitamin D molecules,

wherein said detection agent is an antibody comprising a light chain and a heavy chain, wherein said light chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 16, 17, 18, 19, and 20; and wherein said heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15.

2. The test device of claim 1 , wherein said one or more vitamin D molecules are selected from the group consisting of 25-hydroxyvitamin D 2 , 25-hydroxyvitamin D 3 , 25-hydroxyvitamin D 4 , 25-hydroxyvitamin D 5 , and 1,25-hydroxyvitamin D 3 .

3. The test device of claim 1 , wherein said one or more vitamin D molecules are selected from the group consisting of 25-hydroxyvitamin D 2 and 25-hydroxyvitamin D 3 .

4. The test device of claim 1 , wherein said one or more vitamin D molecules are 25-hydroxyvitamin D 2 and 25-hydroxyvitamin D 3 .

5. The test device of claim 1 , wherein said one or more vitamin D molecules consist of 25-hydroxyvitamin D 2 .

6. The test device of claim 1 , wherein said one or more vitamin D molecules consist of 25-hydroxyvitamin D 3 .

7. The test device of claim 1 , wherein said detection zone further comprises a second region having immobilized thereon an antibody that is capable of binding to said vitamin D binding agent whether or not said vitamin D binding agent is bound to said one or more vitamin D molecules.

8. The test device of claim 1 , wherein said vitamin D binding agent is conjugated to a detection reagent.

9. The test device of claim 1 , wherein said sample application pad, said conjugate pad, and said detection zone are aligned from upstream to downstream along a fluid path through which said biological sample travels.

10. The test device of claim 1 , further comprising a filtering component between said sample application pad and said conjugate pad configured to separate a particulate portion of said biological sample from an aqueous portion of said biological sample.

11. The test device of claim 1 , wherein said detection agent is an antibody comprising a light chain and a heavy chain, wherein:

(i) said light chain comprises the amino acid sequence of SEQ ID NO: 20, and said heavy chain comprises the amino acid sequence of SEQ ID NO: 12; or

(ii) said light chain comprises the amino add sequence of SEQ ID NO: 2, and said heavy chain comprises the amino acid sequence of SEQ ID NO: 13; or

(iii) said light chain comprises the amino add sequence of SEQ ID NO: 19, and said heavy chain comprises the amino acid sequence of SEQ ID NO: 9.

12. A method for detecting one or more vitamin D molecules in a biological sample, comprising:

(a) applying said a biological sample to said sample application pad of said test device of claim 1 ;

(b) applying a chase buffer to said sample application pad; and

(c) detecting said one or more vitamin D molecules using said test device of claim 1 .

13. The method of claim 12 , further comprising quantifying said one or more vitamin D molecules in said biological sample.

14. The method of claim 13 , wherein said quantifying classifies said biological sample as having a sufficient level, an insufficient level, or a deficient level of said one or more vitamin D molecules.

15. The method of claim 13 , wherein said quantifying comprises quantifying a concentration of about 3 ng/ml to about 105 ng/ml of said one or more vitamin D molecules in said biological sample.

16. The method of claim 14 , wherein said sufficient level is at least 30 ng/mL.

17. The method of claim 14 , wherein said insufficient level is at least 10 ng/mL and less than 30 ng/mL.

18. The method of claim 14 , wherein said deficient level is less than 10 ng/mL.

19. The method of claim 13 , wherein said quantifying comprises using: (i) an imaging device to provide an image of said detection zone; and (ii) software on a programmed computer configured to quantify said one or more vitamin D molecules in said biological sample based on said image of said detection zone.

20. The method of claim 12 , wherein said chase buffer comprises reagents to dissociate said one or more vitamin D molecules from vitamin D binding protein.

21. The method of claim 12 , wherein said biological sample is selected from the group consisting of whole blood, serum, plasma, urine, saliva, ocular fluid, spinal fluid, and perspiration.

22. A method for detecting one or more vitamin D levels in a biological sample with a test device configured to perform a binding assay, comprising:

(a) contacting said biological sample with the test device of claim 1 ;

(b) subjecting said biological sample to said binding assay, wherein said binding assay comprises forming a complex between (i) said vitamin D binding agent in said conjugate pad; and (ii) one or more vitamin D molecules in said biological sample;

(c) further exposing said complex to said detection agent immobilized in the detection zone, and detecting said one or more vitamin D molecules by detecting binding of said complex to said detection agent.

23. A kit, comprising:

(a) said test device of claim 1 ; and

(b) written instructions for use of said kit.