Red cell diluent with chelating agent and methods for making and using the same
The present disclosure relates to a RBC solution comprising ethylenediaminetetraacetic acid (EDTA) and methods for making and using the same. The solution can comprise EDTA at a concentration from 3-5 g/L, including 4 g/L. Well plates prepared using the solution can provide a decreased rate of false positive results in automated immunoassays for detecting RBC antibodies in a patient sample.
1 . A method comprising:
preparing a red blood cell (RBC) solution comprising RBCs and a chelating agent, the RBCs comprising antigens to antibodies to be identified in a patient sample;
applying the RBC solution to a structure to cause the RBCs in the solution to be adhered to a surface of the structure; and
applying a drying solution to the structure having the RBCs or fragments of the RBCs adhered to the surface of the structure.
2 . The method of claim 1 , wherein the chelating agent comprises ethylenediaminetetraacetic acid (EDTA).
3 . The method of claim 2 , wherein the RBCs comprise packed RBCs.
4 . The method of claim 3 , wherein the EDTA comprises one or more of disodium EDTA and tetrasodium EDTA.
5 . The method of claim 4 , wherein the solution comprises EDTA at a concentration from about 3 g/L (grams per liter) to about 5 g/L.
6 . The method of claim 5 , wherein the concentration of EDTA is about 4 g/L.
7 . The method of claim 1 , further comprising:
applying a lysing agent to RBCs adhered to the structure to produce the fragments of the RBCs.
8 . The method of claim 7 , wherein the RBC fragments comprise at least one of RBC membranes or RBC stroma.
9 . The method of claim 1 , further comprising, prior to applying the RBC solution:
applying attachment molecules to the structure; and
incubating the attachment molecules so that the attachment molecules adhere to the surface of the structure;
wherein the RBCs or the fragments of the RBCs are bound to respective attachment molecules.
10 . The method of claim 9 , further comprising, after the RBCs or the fragments of the RBCs are bound to respective attachment molecules:
washing the structure, thereby leaving the RBCs or the fragments of the RBCs bound to the respective attachment molecules.
11 . The method of claim 1 , further comprising:
following application of the drying solution, curing the structure having the RBCs or the fragments of the RBCs adhered to the surface of the structure for a period of time.
12 . The method of claim 11 , wherein the period of time is at least three days.
13 . The method of claim 1 , wherein the structure comprises a well, a plate, or a strip.
14 . The method of claim 1 , further comprising:
incorporating the structure having the RBCs or the fragments of the RBCs adhered to the surface thereof into a kit.
15 . The method of claim 14 , wherein the kit comprises:
low ionic strength saline (LISS) solution; and
indicator cells coated with anti-IgG antibody.