Event/sec from single platform flow cytometry for calculation of absolute CD4+ T-lymphocyte and total WBC count in HIV/AIDS patients

Abstract

A single platform flow cytometric analysis using 3 fluorescent-tag monoclonal antibodies and fluorescent microbeads is a standard protocol for measuring CD4+ T-lymphocytes. Whereas dual platform techniques require both flowcytometry and hematological analyzer. In this study absolute CD4+ T-lymphocytes and white blood cell (WBC) counts were calculated from event/sec determined from 193 blood samples from HIV infected individuals or AIDS patients by single platform flow cytometry without fluorescent microbeads comparing to the standard protocol.  The results showed high correlation of absolute CD4+ T-lymphocytes and WBC counts between two methods with R² = 0.9718, p = 0.037 and R² = 0.9196, p < 0.01, respectively. This modified protocol was also evaluated for analytical accuracy (%bias), precision (%CV) and uncertainty of measurements (U_c). Calculation of absolute CD4+ T-lymphocytes and WBC counts from events/sec is simple, rapid and lowered cost comparing to dual platform measurement. This modified method can also reduce the cost of single platform analysis by 18% without decreasing analytical quality.