Störmer M, Cassens U, Kleesiek K, Dreier J
Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Bad Oeynhausen, Germany.
Transfus Med. 2007 Feb;17(1):61-70. doi: 10.1111/j.1365-3148.2006.00710.x.
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.
细菌的生长动力学因血液成分类型而异。在22摄氏度储存时，血小板浓缩物（PCs）似乎比红细胞浓缩物更容易发生细菌增殖。在考虑实施检测策略之前，了解在一系列温度下储存的血液成分中细菌增殖的可能性至关重要。使用实时逆转录聚合酶链反应（RT-PCR），在故意污染的全血（WB）单位获得的血液成分中细菌生长后，确定细菌检测的有效性。培养用作参考方法。将WB接种2个菌落形成单位mL(-1)的表皮葡萄球菌或肺炎克雷伯菌，在室温下保存15小时并进行成分制备。在整个分离过程中，每隔一段时间从每个血液成分中抽取样本。使用自动大容量提取方法提取核酸。15小时的储存显示细菌滴度无明显增加。在红细胞或血浆单位中未检测到细菌生长。肺炎克雷伯菌在混合的PC中显示出快速生长，并且在制备后立即使用RT-PCR可以检测到。表皮葡萄球菌生长缓慢，在分离后24小时被检测到。这些实验表明，最早在PCs制备后24小时取样具有指示性，以尽量减少取样误差。
