Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of Healthcare Quality Promotion, 1600 Clifton Rd., Mail-Stop C16, Atlanta, GA 30333, USA.
Appl Environ Microbiol. 2011 Dec;77(23):8355-9. doi: 10.1128/AEM.05377-11. Epub 2011 Sep 30.
This work was initiated to address the gaps identified by Congress regarding validated biothreat environmental sampling and processing methods. Nine Laboratory Response Network-affiliated laboratories participated in a validation study of a cellulose sponge wipe-processing protocol for the recovery, detection, and quantification of viable Bacillus anthracis Sterne spores from steel surfaces. Steel coupons (645.16 cm(2)) were inoculated with 1 to 4 log(10) spores and then sampled with cellulose sponges (Sponge-Stick; 3M, St. Paul, MN). Surrogate dust and background organisms were added to the sponges to mimic environmental conditions. Labs processed the sponges according to the provided protocol. Sensitivity, specificity, and mean percent recovery (%R), between-lab variability, within-lab variability, and total percent coefficient of variation were calculated. The mean %R (standard error) of spores from the surface was 32.4 (4.4), 24.4 (2.8), and 30.1 (2.3) for the 1-, 2-, and 4-log(10) inoculum levels, respectively. Sensitivities for colony counts were 84.1%, 100%, and 100% for the 1-, 2-, and 4-log(10) inocula, respectively. These data help to characterize the variability of the processing method and thereby enhance confidence in the interpretation of the results of environmental sampling conducted during a B. anthracis contamination investigation.
这项工作的目的是为了满足国会提出的关于验证生物威胁环境采样和处理方法的要求。九个实验室响应网络附属实验室参与了纤维素海绵擦拭处理方案的验证研究，该方案用于从钢表面回收、检测和定量存活炭疽杆菌孢子。用纤维素海绵（Sponge-Stick；3M，圣保罗，MN）对钢片（645.16 cm(2)）进行采样，接种 1 至 4 个对数（10）孢子。在海绵中添加替代灰尘和背景生物，以模拟环境条件。实验室按照提供的方案处理海绵。计算了灵敏度、特异性和平均回收率（%R）、实验室间变异性、实验室内变异性和总变异系数百分比。表面孢子的平均 %R（标准误差）分别为 32.4（4.4）、24.4（2.8）和 30.1（2.3），用于 1、2 和 4 个对数（10）接种水平。菌落计数的灵敏度分别为 84.1%、100%和 100%，用于 1、2 和 4 个对数（10）接种水平。这些数据有助于描述处理方法的变异性，从而增强对炭疽杆菌污染调查中进行的环境采样结果的解释的信心。
