Effect of dilution on sedimentational separation of bacteria from blood

Abstract

Bacteria must be separated from septic whole blood in preparation for rapid antibiotic susceptibility tests. This work improves upon past work isolating bacteria from whole blood by exploring an important experimental factor: Whole blood dilution. Herein, we use the continuity equation to model red blood cell sedimentation and show that overall spinning time decreases as the blood is diluted. We found that the bacteria can also be captured more efficiently from diluted blood, up to approximately 68 ± 8% recovery (95% confidence interval). However, diluting blood both requires and creates extra fluid that end users must handle; an optimal dilution, which maximizes bacteria recovery and minimizes waste, was found to scale with the square root of the whole blood hematocrit. This work also explores a hypothesis that plasma backflow, which occurs as red cells move radially outward, causes bacterial enrichment in the supernatant plasma with an impact proportional to the plasma backflow velocity. Bacteria experiments carried out with diluted blood demonstrate such bacterial enrichment, but not in the hypothesized manner as enrichment occurred only in undiluted blood samples at physiological hematocrit.

Bacterial Extraction and Concentration from Blood through Filtration Processes

“…In addition to direct whole blood lysis, our present study also examined doing blood-lysis filtration to collect bacteria from plasma from whole blood spun upon a previously described centrifugal-sedimentation device [10,[33][34][35][36][37]. A lysis solution is often needed for the plasma recovered after spinning on this device because the device does not separate all RBCs from the resulting plasma.…”