Methods and compositions for reversing platelet clumping
Provided herein are methods and related compositions and kits for dissolution of platelet clumps formed in collected blood samples. The present methods, compositions and kits are useful for improving blood sample quality and blood assay accuracy in a lab setting, especially in preventing misdiagnosis such as pseudothrombocytopenia or pseudoleukocytosis.
1. A method for dissolution of platelet clumps in a blood sample comprising collecting the blood sample in a heparin-containing container, allowing platelets in the blood sample to clump, and contacting the blood sample in the heparin-containing container with a chelator that reduces reactive Ca2+ from the blood sample thereby reversing platelet clumping, wherein the chelator is present at a concentration of less than about 50% by volume or about 60 mM.
2. The method of claim 1 , wherein the heparin is lithium heparin (LiHep).
3. The method of claim 1 , wherein the chelator comprises ethylenediaminetetraacetic acid (EDTA) or a salt thereof.
4. The method of claim 1 , wherein the chelator comprises citric acid or a salt thereof.
5. The method of claim 1 , wherein the contacting is performed by contacting the chelator with the blood sample containing platelet clumps.
6. The method of claim 1 , wherein the chelator comprises citric acid or a salt thereof, and wherein the contacting is performed by contacting an acid citrate dextrose (ACD) solution with the blood sample to a final ACD concentration in the range of about 5% to 20% by volume.
7. The method of claim 1 , wherein after the contacting the method further comprises applying a physical force to the blood sample.
8. The method of claim 7 , wherein the physical force is vortexing, pipetting, sonication, acoustic mixing, and/or mechanical vibrating.
9. The method of claim 1 , wherein the chelator comprises EDTA or a salt of EDTA, and citric acid or a salt of citric acid, and wherein the contacting is performed by contacting EDTA with the blood sample to a final EDTA concentration of about 18 mM and contacting the ACD solution with the blood sample to a final ACD concentration of about 9.7% by volume.
10. The method of claim 1 , wherein the chelator comprises EDTA or a salt of EDTA, and citric acid or a salt of citric acid, and wherein the contacting is performed by contacting EDTA or the salt of EDTA with the blood sample to a final EDTA concentration of about 16.5 mM and contacting the ACD solution with the blood sample to a final ACD concentration of about 17.6% by volume.
11. The method of claim 1 , wherein the chelator comprises EDTA or a salt thereof and the contacting is performed by contacting EDTA or a salt thereof with the blood sample to a final EDTA concentration of no more than about 20 mM.
12. The method of claim 11 , wherein the contacting step is performed by contacting EDTA or a salt thereof with the blood sample to a final EDTA concentration of in the range of about 4 mM to 20 mM EDTA.
13. The method of claim 1 , wherein the method improves accuracy of an assay.
14. The method of claim 13 , wherein the assay is a complete blood count (CBC) assay, a red blood cell (RBC) count assay, a white blood cell (WBC) count assay, a white blood cell (WBC) differential assay, a platelet count assay or a mean platelet volume (MPV) assay.
15. The method of claim 1 , wherein the method prevents a misdiagnosis of a condition.
16. The method of claim 15 , wherein the misdiagnosis is pseudothrombocytopenia or pseudoleukocytosis.
17. A method for dissolution of platelet clumps in a blood sample comprising collecting the blood sample in a heparin-containing container, allowing platelets in the blood sample to clump, and contacting the blood sample in the heparin-containing container with a chelator that reduces reactive Ca2+ from the blood sample thereby reversing or preventing platelet clumping.