Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.
Transfusion. 2011 Sep;51(9):2006-11. doi: 10.1111/j.1537-2995.2011.03090.x. Epub 2011 Mar 10.
In this study the applicability of a 16S rRNA real-time reverse transcriptase polymerase chain reaction (RT-PCR) and a Staphylococcus genus-specific PCR for screening of bacterial contamination in platelet concentrates (PCs) was determined.
A total of 336 sample bags, from PCs that were routinely tested in the BacT/ALERT (bioMérieux), were collected and frozen until testing by the PCR assays. Based on the BacT/ALERT results, 107 PCs were positive and 229 were negative for bacterial growth.
The analytical sensitivity of the 16S rRNA real-time RT-PCR ranged from 5 to 40 colony-forming units (CFUs)/mL. The PCR detected five positive samples, four of which were also positive in the BacT/ALERT. The sensitivity of the test was 3.8%, and the specificity was 99.5%. The analytical sensitivity of the Staphylococcus genus-specific PCR ranged from 5 to 15 CFUs/mL. Thirty-nine units that were BacT/ALERT positive for staphylococci were tested with this PCR. Six samples were positive with the PCR, five of which were also BacT/ALERT positive. The sensitivity of the Staphylococcus genus-specific PCR was 12.8%, and the specificity was 98.8%.
Despite the rapid availability of results compared to the BacT/ALERT, the analytical sensitivity of a generic or specific PCR assay is not high enough to be an alternative for the BacT/ALERT when PCs are screened on the day of production.
本研究旨在确定 16S rRNA 实时逆转录聚合酶链反应（RT-PCR）和葡萄球菌属特异性 PCR 用于筛选血小板浓缩物（PC）中细菌污染的适用性。
共收集 336 个样本袋，这些样本来自常规在 BacT/ALERT（bioMérieux）中进行检测的 PC，并在进行 PCR 检测前进行冷冻保存。根据 BacT/ALERT 的结果，107 个 PC 对细菌生长呈阳性，229 个 PC 呈阴性。
16S rRNA 实时 RT-PCR 的分析灵敏度范围为 5 至 40 个菌落形成单位（CFU）/mL。PCR 检测到五个阳性样本，其中四个在 BacT/ALERT 中也呈阳性。该检测的灵敏度为 3.8%，特异性为 99.5%。葡萄球菌属特异性 PCR 的分析灵敏度范围为 5 至 15 CFU/mL。对 39 个 BacT/ALERT 检测到葡萄球菌阳性的单位进行了该 PCR 检测。有 6 个样本 PCR 检测呈阳性，其中 5 个样本在 BacT/ALERT 中也呈阳性。葡萄球菌属特异性 PCR 的灵敏度为 12.8%，特异性为 98.8%。
尽管与 BacT/ALERT 相比，结果可以更快获得，但通用或特异性 PCR 检测的分析灵敏度还不够高，无法替代 BacT/ALERT 在 PC 生产当天进行筛选。
