A flow-through cell counting assay for point-of-care enumeration of CD4 T-cells

CD4 T-cell count is a priority for staging HIV disease and guiding clinical management as part of HIV care. Conventional CD4 T-cell enumeration methods based on flow cytometry are expensive, require well-trained personnel, and are challenging to use in rural, resource-scarce areas. A simple CD4 T-cell count test that can be used at point-of care, the Flow-Through cell Counting Assay (FTCA), is described in this article. The FTCA is based on the use of: 1) a special membrane that selectively retains white blood cells (WBCs); 2) a sample delivery system; and 3) optical signal detection. To show the feasibility of the FTCA, a proof-of-concept prototype of the FTCA cassette and digital camera or handheld reflectance meter were used for obtaining quantitative assay results within 30 min. The results show that the FTCA allows for quantitative enumeration of CD4 T-cells in the clinically relevant range of CD4 T-cell concentrations. The advantages of the FTCA technology, including simplicity, short analysis time, and portability, suggest that FTCA has great potential for use in clinical practice and wide applicability for other cell-based diagnostic tests.