Keil Shawn D, Hovenga Nick, Gilmour Denise, Marschner Susanne, Goodrich Raymond
Terumo BCT, Scientific Affairs;
Terumo BCT, Scientific Affairs.
J Vis Exp. 2015 Aug 24(102):e52820. doi: 10.3791/52820.
Contamination of platelet units by bacteria has long been acknowledged as a significant transfusion risk due to their post-donation storage conditions. Products are routinely stored at 22 °C on an agitating shaker, a condition that can promote bacterial growth. Although the total number of bacteria believed to be introduced into a platelet product is extremely low, these bacteria can multiply to a very high titer prior to transfusion, potentially resulting in serious adverse events. The aim of this study was to evaluate a riboflavin based pathogen reduction process against a panel of bacteria that have been identified as common contaminants of platelet products. This panel included the following organisms: S. epidermidis, S. aureus, S. mitis, S. pyogenes, S. marcescens, Y. enterocolitica, B. neotomae, B. cereus, E. coli, P. aeruginosa and K. pneumoniae. Each platelet unit was inoculated with a high bacterial load and samples were removed both before and after treatment. A colony forming assay, using an end point dilution scheme, was used to determine the pre-treatment and post-treatment bacterial titers. Log reduction was calculated by subtracting the post-treatment titer from the pre-treatment titer. The following log reductions were observed: S. epidermidis 4.7 log (99.998%), S. aureus 4.8 log (99.998%), S. mitis 3.7 log (99.98%), S. pyogenes 2.6 log (99.7%), S. marcescens 4.0 log (99.99%), Y. enterocolitica 3.3 log (99.95%), B. neotomae 5.4 log (99.9996%), B. cereus 2.6 log (99.7%), E. coli ≥5.4 log (99.9996%), P. aeruginosa 4.7 log (99.998%) and K. pneumoniae 2.8 log (99.8%). The results from this study suggest the process could help to lower the risk of severe adverse transfusion events associated with bacterial contamination.
由于血小板制品在采集后的储存条件,其受细菌污染长期以来一直被认为是一种重大的输血风险。制品通常在22℃下于振荡摇床上储存,这种条件会促进细菌生长。尽管据信引入血小板制品中的细菌总数极低,但这些细菌在输血前可繁殖至非常高的滴度,有可能导致严重不良事件。本研究的目的是评估一种基于核黄素的病原体灭活工艺对一组已被确定为血小板制品常见污染物的细菌的效果。该组包括以下微生物:表皮葡萄球菌、金黄色葡萄球菌、缓症链球菌、化脓性链球菌、粘质沙雷氏菌、小肠结肠炎耶尔森菌、新托马芽孢杆菌、蜡样芽孢杆菌、大肠杆菌、铜绿假单胞菌和肺炎克雷伯菌。每个血小板制品接种高细菌载量,并在处理前后取样。采用终点稀释方案的菌落形成试验用于确定处理前和处理后的细菌滴度。通过从处理前滴度中减去处理后滴度来计算对数减少率。观察到以下对数减少率:表皮葡萄球菌4.7对数(99.998%)、金黄色葡萄球菌4.8对数(99.998%)、缓症链球菌3.7对数(99.98%)、化脓性链球菌2.6对数(99.7%)、粘质沙雷氏菌4.0对数(99.99%)、小肠结肠炎耶尔森菌3.3对数(99.95%)、新托马芽孢杆菌5.4对数(99.9996%)、蜡样芽孢杆菌2.6对数(99.7%)、大肠杆菌≥5.4对数(99.9996%)、铜绿假单胞菌4.7对数(99.998%)和肺炎克雷伯菌2.8对数(99.8%)。本研究结果表明该工艺有助于降低与细菌污染相关的严重不良输血事件的风险。