Open AccessRapid, sensitive detection of bacteria in platelet samples with Fountain Flow Cytometry
Author(s) -
Johnson Paul,
Moriwaki Mika,
Johnson Joseph
Publication year - 2017
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22115
Subject(s) - flow cytometry , serial dilution , bacteria , chromatography , enumeration , fluorescence , platelet , stain , microbiology and biotechnology , chemistry , biology , staining , medicine , pathology , immunology , mathematics , genetics , alternative medicine , physics , combinatorics , quantum mechanics
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 R 2 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.