Transfusion and apheresis science : official journal of the World Apheresis Association : official journal of the European Society for Haemapheresis · 2013
Three different bacterial detection systems for platelet concentrates under inter-laboratory conditions.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
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GuoHui Bian, Chunhui Yang, Miao He, Hong Wang, Jiaxin Liu, Ye Cao, Hong Yang, Zhong Liu, Wuping Li
Аннотация
BACKGROUND: A variety of screening methods are currently used worldwide in order to decrease the risk of transfusion-transmitted sepsis and improve the safety of PCs.
METHODS/MATERIALS: PCs inoculated with five different transfusion-relevant species of bacteria at concentrations of 1, 10, and 100 colony-forming units (CFU)ml(-1) were stored at 22°C for 7 days. Flow cytometry (FACS), BacT/Alert automated culturing, and a quantitative real-time PCR (Q-PCR) were then used to detect the presence of bacteria in samples prepared from these PCs.
RESULTS: At the initial spiking concentrations of 1, 10, and 100 CFU ml(-1), Q-PCR detected all five bacterial species tested. Screening with the BacT/Alert culture-based system allowed bacterial detection (inoculated on day 0) within a mean time of 15.13 h for all three spiking concentrations. Using FACS, positive signals were obtained for all three concentrations of Escherichia coli and Bacillus cereus on day 1 and for initial spiking concentrations of Pseudomonas aeruginosa and Staphylococcus aureus of 1 CFU ml(-1) on day 2. For Staphylococcus epidermidis, detection of an initial inoculum of 1 CFU ml(-1) was possible only beginning on day 6.
CONCLUSION: This study shows that under standard laboratory conditions the sensitivity of FACS in the detection of bacterial contamination of PCs was lower than that of either the BacT/Alert automated culturing method or Q-PCR.
METHODS/MATERIALS: PCs inoculated with five different transfusion-relevant species of bacteria at concentrations of 1, 10, and 100 colony-forming units (CFU)ml(-1) were stored at 22°C for 7 days. Flow cytometry (FACS), BacT/Alert automated culturing, and a quantitative real-time PCR (Q-PCR) were then used to detect the presence of bacteria in samples prepared from these PCs.
RESULTS: At the initial spiking concentrations of 1, 10, and 100 CFU ml(-1), Q-PCR detected all five bacterial species tested. Screening with the BacT/Alert culture-based system allowed bacterial detection (inoculated on day 0) within a mean time of 15.13 h for all three spiking concentrations. Using FACS, positive signals were obtained for all three concentrations of Escherichia coli and Bacillus cereus on day 1 and for initial spiking concentrations of Pseudomonas aeruginosa and Staphylococcus aureus of 1 CFU ml(-1) on day 2. For Staphylococcus epidermidis, detection of an initial inoculum of 1 CFU ml(-1) was possible only beginning on day 6.
CONCLUSION: This study shows that under standard laboratory conditions the sensitivity of FACS in the detection of bacterial contamination of PCs was lower than that of either the BacT/Alert automated culturing method or Q-PCR.