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Abstract
The development of point-of-care, cost-effective, and easy-to-use assays for the accurate counting of CD4+ T cells remains an important focus for HIV-1 disease management. The CD4+ T cell count provides an indication regarding the overall success of HIV-1 treatments. The CD4+ T count information is equally important for both resource-constrained regions and areas with extensive resources. Hospitals and other allied facilities may be overwhelmed by epidemics or other disasters. An assay for a physician’s office or other home-based setting is becoming increasingly popular. We have developed a technology for the rapid quantification of CD4+ T cells. A double antibody selection process, utilizing anti-CD4 and anti-CD3 antibodies, is tested and provides a high specificity. The assay utilizes a microfluidic chip coated with the anti-CD3 antibody, having an improved antibody avidity. As a result of enhanced binding, a higher flow rate can be applied that enables an improved channel washing to reduce non-specific bindings. A wide-field optical imaging system is also developed that provides the rapid quantification of cells. The designed optical setup is portable and low-cost. An ImageJ-based program is developed for the automatic counting of CD4+ T cells. We have successfully isolated and counted CD4+ T cells with high specificity and efficiency greater than 90%.
Highlights
There is a need to develop accurate cell quantification assays to achieve early-stage disease detection, treatment, and monitoring
We describe an improved microfluidic chip process that separates a sparse number of CD4+ T lymphocytes from whole blood samples
Anti-CD3 antibody, UCHT1(biotin), part number ab191112 was obtained from Abcam, and it was used to coat the bottom glass substrate of the microchip
Summary
There is a need to develop accurate cell quantification assays to achieve early-stage disease detection, treatment, and monitoring. As a result of these advances, several sophisticated and advanced laboratory-based devices have been tested and approved for accurate cell quantification purposes. Such devices require well-trained personnel in well-equipped laboratories. There is an unmet need to develop cost-effective, easy-to-use, and rapid disease diagnostic devices at the pointof-care settings, POC. Devices developed based on these guidelines would be beneficial for both resource-enabled and resource-limited countries. This lack of adequate resources applies to physicians’
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