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Spinning Disk as a Novel Method for Isolation of Plasma Prior to Cell-free Ribonucleic Acid (cfRNA) Extraction and Utilization

Year: 2023

Presenter Name: Mae-Lynn Hutchinson

The utilization of cell-free ribonucleic acids (cfRNA), such as found in a liquid biopsy, has revolutionized the early detection and monitoring of some health conditions. Such conditions include pregnancy complications, viral infections, and cancer. cfRNA is typically isolated through a rapid centrifugation procedure; however, the use of an inexpensive spinning disk has been considered in some locations and applications. Briefly, this approach involves applying a centrifugal field to a hollow spinning disk to remove cellular material, with RNA remaining, from the blood sample in ~2 minutes.To examine the effectiveness of these two methods, blood was collected from healthy donors into pairs of Streck RNA Complete BCT tubes. Sample hematocrit was measured and samples were diluted with phosphate buffered saline (PBS) to a consistent hematocrit of 32. From each pair of samples, one control sample underwent the standard centrifugation process, while the other sample underwent the spinning disk procedure to collect plasma. Plasma hemolysis was evaluated using spectrophotometry according to the Harboe method with the Allen correction prior to cfRNA isolation. Following cfRNA isolation, qRT-PCR of the GAPDH and beta-actin transcripts was performed to evaluate quality and quantity of the cfRNA recovered by each method. We found that cfRNA yield from the centrifugation and the spinning disk methods did not differ significantly (p = 0.077), though there was significantly more hemolysis in the plasma from the spinning disk method (p = 0.004). Our findings suggest that the spinning disk could expand possibilities for rapid, effective, high-quality treatment and monitoring of patients living in areas where a full-size high speed centrifuge is not accessible since the disk is much smaller and cheaper than other platforms.
University / Institution: Brigham Young University
Type: Poster
Format: In Person
Presentation #A67
SESSION A (9:00-10:30AM)
Area of Research: Health & Medicine
Faculty Mentor: Bill Pitt