Method for stabilizing hemoglobin and reagents for performing the same
A stool resuspension solution comprising a hemoglobin stabilization reagent is provided. In some embodiments, the hemoglobin stabilization reagent may be an osmolyte, a polyvalent cation, a sugar or polysaccharide and, optionally, a polyvalent cation, a protoporphyrin, or an HRP stabilization component and, optionally, a polyvalent cation. A method of stabilizing hemoglobin in a stool sample in the solution is also provided, as well as a sample collection device containing the solution.
1. A composition comprising: a stool resuspension solution comprising:
a buffer; and
a protoporphyrin complexed with Cr 3+ or Co 3+ , wherein the concentration of the protoporphyrin complexed with Cr 3+ is 1.25-5 μM and the concentration of the protoporphyrin complexed with Co 3+ is 2.5-10 μM,
wherein the protoporphyrin complexed with Cr 3+ or Co 3+ of is effective to stabilize hemoglobin.
2. The composition of claim 1 , wherein the protoporphyrin is protoporphyrin IX complexed with Cr 3+ and the concentration of protoporphyrin IX complexed with Cr 3+ is 1.5-4.5 μM.
3. The composition of claim 1 , wherein the protoporphyrin is protoporphyrin IX complexed with Co 3+ and the concentration of protoporphyrin IX complexed with Co 3+ is 3-5 μM.
4. The composition of claim 1 , wherein the solution has a pH in the range of pH 6.5 to pH 7.4.
5. The composition of claim 2 , wherein the pH of the solution is in the range of pH 6.9 to pH 7.4.
6. The composition of claim 3 , wherein the pH of the solution is in the range of pH 6.5 to pH 7.4.
7. The composition of claim 1 , wherein a stool sample is suspended in the solution.
8. The composition of claim 1 , wherein the buffer is Tris buffer and the solution further comprises bovine serum albumen, polysorbate 20, sodium azide, sodium chloride, ethylenediaminetetraacetic acid, and gentamicin.
9. A method of stabilizing hemoglobin in a stool sample, comprising:
combining the stool sample with a stool resuspension solution comprising a buffer and a protoporphyrin complexed with Cr 3+ or Co 3± ; to produce a suspension; and
maintaining the suspension for a period of time,
wherein the concentration of the protoporphyrin complexed with Cr 3+ is 1.25-5 μM and the concentration of the protoporphyrin complexed with Co 3+ is 2.5-10 μM, and
wherein the protoporphyrin complexed with Cr 3+ or Co 3+ increases stability of hemoglobin.
10. The method of claim 9 , wherein the method comprises shipping the suspension to a remote location.
11. A method of analyzing a stool sample, comprising:
(a) receiving, from a remote location, a composition comprising:
(i.) a stool sample; and
(ii.) a stool resuspension solution comprising a buffer and a protoporphyrin complexed with Cr 3+ or Co 3+ ,
wherein the concentration of the protoporphyrin complexed with Cr 3+ is 1.25-5 μM and the concentration of the protoporphyrin complexed with Co 3+ is 2.5-10 μM, and
wherein the protoporphyrin complexed with Cr 3+ or Co 3+ increases stability of hemoglobin;
wherein the stool sample is suspended in the solution; and
(b) measuring the amount of hemoglobin in the composition.
12. The method of claim 11 , further comprising measuring the amount of one or more colorectal cancer tumor markers in the sample.
13. The method of claim 11 , wherein the protoporphyrin is protoporphyrin IX complexed with Cr 3+ and the concentration of protoporphyrin IX complexed with Cr 3+ is 1.5-4.5 μM.
14. A sample collection device comprising:
a sample collection container having an open end;
a stool resuspension solution comprising a buffer and a protoporphyrin complexed with Cr 3+ or Co 3+ , wherein the concentration of the protoporphyrin complexed with Cr 3+ is 1.25-5 μM and the concentration of the protoporphyrin complexed with Co 3+ is 2.5-10 μM, and wherein the protoporphyrin complexed with Cr 3+ or Co 3+ increases stability of is effective to stabilize hemoglobin; and
a stool sampling rod comprising a distal beveled tip for scooping and/or scraping a sample of stool,
wherein the distal end of the sampling rod is dimensioned to be inserted into the sample collection container and the proximal end of the sampling rod is adapted to connect with the open end of the sample collection container, thereby sealing distal end of the sampling rod within the device.
15. The sample collection device of claim 14 , wherein the sample collection container and sampling rod connect via a screw fit.
16. The sample collection device of claim 14 , wherein the protoporphyrin is protoporphyrin IX complexed with Cr 3+ and the concentration of protoporphyrin IX complexed with Cr 3+ is 1.5-4.5 μM.
17. The sample collection device of claim 14 , wherein the protoporphyrin is protoporphyrin IX complexed with Co 3+ and the concentration of protoporphyrin IX complexed with Co 3+ is 3-5 μM.
18. The method of claim 11 , wherein the protoporphyrin is protoporphyrin IX complexed with Co 3+ and the concentration of protoporphyrin IX complexed with Co 3+ is 3-5 μM.
19. The sample collection device of claim 14 , wherein the stool resuspension solution is within the container.
20. The composition of claim 8 , wherein the concentration of protoporphyrin IX complexed with Cr 3+ is 2.5 μM.
21. The composition of claim 20 , wherein the pH of the composition is 6.9-7.4.
22. The composition of claim 8 , wherein the concentration of protoporphyrin IX complexed with Co 3+ is 4 μM.
23. The composition of claim 22 , wherein the pH of the composition is 6.9-7.4.