Biochemical Science Division, Material Measurements Laboratory, National Institute of Standards and Technology, Gaithersburg, Maryland, USA.
Appl Environ Microbiol. 2012 Aug;78(16):5872-81. doi: 10.1128/AEM.00873-12. Epub 2012 Jun 15.
Environmental sampling for microbiological contaminants is a key component of hygiene monitoring and risk characterization practices utilized across diverse fields of application. However, confidence in surface sampling results, both in the field and in controlled laboratory studies, has been undermined by large variation in sampling performance results. Sources of variation include controlled parameters, such as sampling materials and processing methods, which often differ among studies, as well as random and systematic errors; however, the relative contributions of these factors remain unclear. The objective of this study was to determine the relative impacts of sample processing methods, including extraction solution and physical dissociation method (vortexing and sonication), on recovery of Gram-positive (Bacillus cereus) and Gram-negative (Burkholderia thailandensis and Escherichia coli) bacteria from directly inoculated wipes. This work showed that target organism had the largest impact on extraction efficiency and recovery precision, as measured by traditional colony counts. The physical dissociation method (PDM) had negligible impact, while the effect of the extraction solution was organism dependent. Overall, however, extraction of organisms from wipes using phosphate-buffered saline with 0.04% Tween 80 (PBST) resulted in the highest mean recovery across all three organisms. The results from this study contribute to a better understanding of the factors that influence sampling performance, which is critical to the development of efficient and reliable sampling methodologies relevant to public health and biodefense.
环境微生物污染物采样是卫生监测和风险特征描述实践的关键组成部分,广泛应用于多个领域。然而,由于采样性能结果存在较大差异,现场和受控实验室研究中的表面采样结果的可信度受到了影响。变异的来源包括采样材料和处理方法等受控参数,这些参数在研究中经常不同,以及随机和系统误差;然而,这些因素的相对贡献仍不清楚。本研究的目的是确定样品处理方法(包括提取溶液和物理分离方法[涡旋和超声])对直接接种拭子中革兰氏阳性(蜡样芽孢杆菌)和革兰氏阴性(泰国伯克霍尔德菌和大肠杆菌)细菌回收的相对影响。这项工作表明,目标生物对提取效率和回收精度的影响最大,这是通过传统的菌落计数来衡量的。物理分离方法(PDM)的影响可以忽略不计,而提取溶液的作用则取决于生物体。然而,总体而言,使用含 0.04%吐温 80 的磷酸盐缓冲盐水(PBST)从拭子中提取生物体的效果在所有三种生物体中均能获得最高的平均回收率。本研究的结果有助于更好地了解影响采样性能的因素,这对于开发与公共卫生和生物防御相关的高效可靠采样方法至关重要。