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Rapid, sensitive detection of bacteria in platelet samples with Fountain Flow Cytometry.
Journal of Clinical Laboratory Analysis 2017 November
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.
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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