Laser incubation for the rapid detection of red cell alloantibodies in human blood samples.

BACKGROUND AND OBJECTIVES: Pre-transfusion antibody screening requires the detection and identification of immunoglobulin G (IgG) antibodies against red blood cells (RBCs). Using the indirect antiglobulin test (IAT), plasma-RBC solutions are incubated at 37°C in gel cards, typically by heating block technology. Here, we apply the newly developed laser incubation method to detect RBC alloantibodies in the plasma from human donors. MATERIALS AND METHODS: Donated human plasma samples (N = 128) containing clinically significant IgG antibodies directed against Rh (D, C, c, Cw and E), Kell (K and Kpa ), Duffy (Fya and Fyb ), Kidd (Jka ) and MNS (S) blood group system antigens were tested by the indirect antiglobulin test (IAT). Samples were heated to 37°C by infrared laser (980 nm) for incubations of up to 5 min. Samples were also incubated in a heating block for comparison. RESULTS: When heating by laser, the presence of an alloantibody is detected after only a 1-min incubation for 96% of samples. No samples required longer than 3 min of laser incubation in order to detect the antibody. For all samples, incubation by laser gave the same or stronger result within 5 min. No samples required longer than 5 min to achieve an equivalent result to that of the 5-min heating block incubation. The laser was not found to damage cells or antibodies. CONCLUSION: Laser incubation provides comparable results in shorter time frames than the heating block. Laser incubation can rapidly detect even very weak antibodies.

Photothermal incubation of red blood cells by laser for rapid pre-transfusion blood group typing.

Safe blood transfusion requires compatibility testing of donor and recipient to prevent potentially fatal transfusion reactions. Detection of immunoglobulin G (IgG) antibodies requires incubation at 37 °C, often for up to 15 minutes. Current incubation technology predominantly relies on slow thermal-gradient dependent conduction. Here, we present rapid optical heating via laser, where targeted illumination of a blood-antibody sample in a diagnostic gel card is converted into heat, via photothermal absorption. Our laser-incubator heats the 75 µL blood-antibody sample to 37 °C in under 30 seconds. We show that red blood cells act as photothermal agents under near-infrared laser incubation, triggering rapid antigen-antibody binding. We detect no significant damage to the cells or antibodies for laser incubations of up to fifteen minutes. We demonstrate laser-incubated immunohaematological testing to be both faster and more sensitive than current best practice - with clearly positive results seen from laser incubations of just 40 seconds.

Nanocellulose Hydrogel for Blood Typing Tests.

The gel test is the most prevalent method for the forward and reverse blood typing tests. It relies on the controlled centrifugation of red blood cells (RBCs) and antibodies through a gel column. This noncontinuous matrix is currently based on microbeads that often lack sensitivity. For the first time, nanocellulose hydrogel is demonstrated as a sustainable and reliable medium for gel-based blood typing diagnostics. Gels with a minimum of 0.3 wt % TEMPO-oxidized cellulose nanofibers (0.92 mmol/g of carboxyl content) separate agglutinated and individual RBCs in the forward test. The addition of glycine is able to balance the osmotic pressure and reduce hemolysis to 5%, while retaining the electrostatic repulsion responsible for the gel network structure and its rheological properties. For the reverse typing, cellulose nanofibers are chemically cross-linked with hexamethylenediamine (HMDA), increasing the gel yield point 8-fold. Sodium chloride is added to achieve the osmolality found in the human plasma and limit cell lysis to 15%, without affecting the gel colloidal stability. Nanocellulose hydrogel constitutes a performant, low cost, and green soft material, providing clear and well-defined results for both blood grouping tests.