Impact of Processing Method on Recovery of Bacteria from Wipes Used in Biological Surface Sampling
Impact of Processing Method on Recovery of Bacteria from Wipes Used in Biological Surface Sampling
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DOI：
10.1128/aem.00873-12
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发表时间：
2012-08-01
影响因子：
4.4
通讯作者：
Morrow, Jayne B.
中科院分区：
文献类型：
--
作者：
Downey, Autumn S.;Da Silva, Sandra M.;Morrow, Jayne B.
微生物污染物的环境采样是卫生监测和风险表征实践的关键组成部分，在不同的应用领域使用。然而，在实地和受控实验室研究中，对地面抽样结果的信心已因抽样性能结果的巨大差异而受到破坏。变异的来源包括受控参数，如采样材料和处理方法，这些在不同的研究中往往不同，以及随机和系统误差；然而，这些因素的相对作用仍不清楚。本研究的目的是确定样品处理方法，包括提取溶液和物理解离法（旋涡和超声）对从直接接种的湿巾中回收革兰氏阳性（蜡样芽孢杆菌）和革兰氏阴性（泰国伯克霍氏菌和大肠杆菌）细菌的相对影响。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，以传统的菌落计数来衡量。物理解离法（PDM）的影响可以忽略不计，而萃取液的影响是生物体依赖的。然而，总体而言，使用含0.04% Tween 80 （PBST）的磷酸盐缓冲盐水从湿巾中提取生物时，三种生物的平均回收率最高。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的采样方法至关重要。
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.