Evaluation of a Protein Microarray Method for Immuno‐Typing Erythrocytes in Whole Blood

Abstract

All donor blood samples must be tested pre-transfusion to determine the blood type of donor erythrocytes, based on the ABO typing system. Current methods of testing are well characterised, but require a number of processing steps prior to analysis. In addition, standard testing protocols require additional assays such as hepatitis C and HIV testing be performed separately. We describe and evaluate a protein microarray platform for ABO blood typing that has the potential to be a simple reliable high throughput method, with the added capability for the integration of other important pre-transfusion tests. Sixty seven donor blood samples were incubated on microarrays printed with multiple spotted replicates of blood type antigen specific antibodies. We utilised a hold-out cross validation approach, combined with Receiver Operator Characteristic (ROC) curves to define thresholds within which a sample could be defined as being of a particular blood type. The threshold values from the ROC curve analysis demonstrated an excellent ability to accurately separate samples based on ABO blood type. The results obtained when the thresholds from the training sets were applied to test sets were also very encouraging, with misclassified samples being present in only 2 of the training sets and a mean classification error of 4.28%. When the mean thresholds were applied to the 67 donor samples, 95.5% were correctly blood typed (64 of 67 samples). We have demonstrated the ability of our protein microarray platform to successfully and accurately type human whole blood samples. We believe that this flexible platform provides a strong basis for an integrated approach for combined blood typing and pathogen testing in human whole blood.

Advances in Microfluidics for Single Red Blood Cell Analysis

“…Immuno-phenotyping applications [335][336][337][338] Detection of complex mixtures and antigens, pre-transfusion testing, serodiagnosis, and genotyping Futuristic assaying methodology, device miniaturization, and combined testing Mesh filtration and Erythrocyte deformability analysis [339] RBC rheology through a nickel mesh filter…”

“…Research has been significantly progressing in single-cell analysis with RBC/Erythrocyte trapping techniques. For example, diagnostic microarrays that can analyze protein samples via antibody-based microarrays have been shown as effective, time-saving, and labelfree techniques for screening the RBC surface antigens and RBC phenotyping [335,336]. Sero-diagnostics from donor blood samples also holds potential for blood phenotyping, pathogenic infections, and blood typing from erythrocytes [337].…”

“…The very first complication is the performance of cells on microarray surfaces, or the prospects for the enhancement of material surface interactions [335]. Advances in miniaturized and instantaneous Point-Of-Care (POC) systems are in desperate needs [336]. Assaying and identifying multiple blood types within a single device is an effective and time-saving method for analyses, and this could be included in the list of future developments [337].…”

Multicenter evaluation of a high‐throughput microarray platform for extensive red blood cell phenotyping and antibody screening

“…The principle of a microarray‐based method for immune‐typing RBCs in whole blood was previously described 5 . We report the performances of a CE‐marked high‐throughput platform (analyzer and reagents) for blood donor grouping and ABS compared with routinely‐used platforms and manual systems in three blood donation laboratories.…”