Novel flow cytometry-based screening for bacterial contamination of donor platelet preparations compared with other rapid screening methods.
BACKGROUND
Bacterial contamination is the major infectious hazard associated with transfusion of platelet preparations (PLTs). Routine testing for bacterial contamination in PLTs has become common, but transfusion-transmitted bacterial sepsis has not been eliminated. Here, we describe a novel flow cytometry-based method for point-of-issue screening of PLTs for bacterial contamination.


METHODS
We used the BactiFlow flow cytometer to detect and count bacteria based on esterase activity in viable cells. We compared the assay to incubation (BacT/Alert culture system) and rapid nucleic acid-based or immunoassay (reverse transcription PCR, Pan Genera Detection) methods.


RESULTS
We established a protocol for bacterial screening of PLTs consisting of enzymatic digestion and centrifugal filtration for the elimination of viable platelets and selective labeling of bacteria with fluorescent esterase substrate (ChemChrome V23). Results from the BactiFlow showed an excellent correlation (r = 0.9923 E. coli, r = 0.9736 S. epidermidis) to traditional plate count results. The lower detection limit of the assay was determined to be 150 CFU/mL, and the time to result was <1 h.


CONCLUSIONS
Our study demonstrates that BactiFlow flow cytometry is suitable for rapid screening of PLTs for bacterial contamination and fulfils the requirements for a point-of-issue testing of PLTs with acceptable time to result, specificity, sensitivity, and cost.
Dreier
Jens
J
Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Bad Oeynhausen, Germany. jdreier@hdz-nrw.de
Vollmer
Tanja
T
Kleesiek
Knut
K
D001419:Bacteria / Q000254:growth & development / Q000302:isolation & purification*; D001792:Blood Platelets / Q000382:microbiology*; D015169:Colony Count, Microbial / Q000191:economics / Q000379:methods*; D005434:Flow Cytometry / Q000191:economics / Q000379:methods*; D006801:Humans; D012680:Sensitivity and Specificity; D013997:Time Factors