Detection of bacteria in platelet concentrates prepared from spiked single donations using cultural and molecular genetic methods.
Bacteria show differences in their growth kinetics depending on the type of blood component. On to storage at 22 degrees C, platelet concentrates (PCs) seem to be more prone to bacterial multiplication than red cell concentrates. Knowledge of the potential for bacterial proliferation in blood components, which are stored at a range of temperatures, is essential before considering implementation of a detection strategy. The efficacy of bacterial detection was determined, using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), following bacterial growth in blood components obtained from a deliberately contaminated whole-blood (WB) unit. Cultivation was used as the reference method. WB was spiked with 2 colony-forming units mL(-1)Staphylococcus epidermidis or Klebsiella pneumoniae, kept for 15 h at room temperature and component preparation was processed. Samples were drawn, at intervals throughout the whole separation process, from each blood component. Nucleic acids were extracted using an automated high-volume extraction method. The 15-h storage revealed an insignificant increase in bacterial titre. No bacterial growth was detected in red blood cell or plasma units. K. pneumoniae showed rapid growth in the pooled PC and could be detected immediately after preparation using RT-PCR. S. epidermidis grew slowly and was detected 24 h after separation. These experiments show that sampling is indicative at 24 h after preparation of PCs at the earliest to minimize the sampling error.
Störmer
M
M
Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Bad Oeynhausen, Germany. jdreier@hdz-nrw.de
Cassens
U
U
Kleesiek
K
K
Dreier
J
J
D000332:Aerobiosis; D001431:Bacteriological Techniques / Q000295:instrumentation; D001769:Blood / Q000382:microbiology; D001782:Blood Donors; D001792:Blood Platelets / Q000382:microbiology*; D002498:Centrifugation; D004269:DNA, Bacterial / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D008212:Lymphocyte Depletion; D008280:Magnetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013212:Staphylococcus epidermidis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D013696:Temperature
D004269:DNA, Bacterial