Paul Johnson1,2, Mika Moriwaki3, Joseph Johnson4. 1. Department of Physics and Astronomy, University of Wyoming, Laramie, WY, USA. 2. SoftRay Inc, Laramie, WY, USA. 3. Molecular Medicine Program, University of Utah, Salt Lake City, UT, USA. 4. College of Osteopathic Medicine of the Pacific, Western University of Health Sciences, Pomona, CA, USA.
Abstract
BACKGROUND: There is a current need to develop a technique for bacterial screening of platelet donations that is more rapid, sensitive, and economical than alternatives. The objective of this research was to perform a pilot test of the viability of Fountain Flow Cytometry (FFC), for the rapid and sensitive detection of bacteria in platelet donations. METHODS: Platelet samples were inoculated with serial dilutions of five selected bacterial strains. Samples were then centrifuged, reconstituted in buffer, and stained with a live/dead bacterial stain cocktail. The resulting aqueous sample was measured by FFC, in which the sample passed as a stream in front of an LED, which excited the fluorescent labels. Fluorescence was detected with a digital camera as the sample flowed toward it. RESULTS: Fountain Flow Cytometry enumeration yielded results that were linear with bacterial concentration, having an R2 of ≥0.98 with a detection efficiency of 92%±3%. Measurements of uninoculated samples showed a false-positive detection rate at ~400 colony forming units (CFU)/mL. Detection of bacterial concentrations in platelets above this threshold can be made in ~15 minutes, including sample preparation time. CONCLUSION: This pilot study supports the efficacy of FFC for the rapid and sensitive screening of platelet donations for bacteria.
BACKGROUND: There is a current need to develop a technique for bacterial screening of platelet donations that is more rapid, sensitive, and economical than alternatives. The objective of this research was to perform a pilot test of the viability of Fountain Flow Cytometry (FFC), for the rapid and sensitive detection of bacteria in platelet donations. METHODS: Platelet samples were inoculated with serial dilutions of five selected bacterial strains. Samples were then centrifuged, reconstituted in buffer, and stained with a live/dead bacterial stain cocktail. The resulting aqueous sample was measured by FFC, in which the sample passed as a stream in front of an LED, which excited the fluorescent labels. Fluorescence was detected with a digital camera as the sample flowed toward it. RESULTS: Fountain Flow Cytometry enumeration yielded results that were linear with bacterial concentration, having an R2 of ≥0.98 with a detection efficiency of 92%±3%. Measurements of uninoculated samples showed a false-positive detection rate at ~400 colony forming units (CFU)/mL. Detection of bacterial concentrations in platelets above this threshold can be made in ~15 minutes, including sample preparation time. CONCLUSION: This pilot study supports the efficacy of FFC for the rapid and sensitive screening of platelet donations for bacteria.
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