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影响生物表面采样擦拭物中孢子回收的参数。

Parameters affecting spore recovery from wipes used in biological surface sampling.

机构信息

Biochemical Science Division, Chemical Science Technology Laboratory, National Institute of Standards and Technology, 100 Bureau Dr., Mail Stop 8312, Gaithersburg, MD 20899-8312, USA.

出版信息

Appl Environ Microbiol. 2011 Apr;77(7):2374-80. doi: 10.1128/AEM.01932-10. Epub 2011 Feb 4.

Abstract

The need for the precise and reliable collection of potential biothreat contaminants has motivated research in developing a better understanding of the variability in biological surface sampling methods. In this context, the objective of this work was to determine parameters affecting the efficiency of extracting Bacillus anthracis Sterne spores from commonly used wipe sampling materials and to describe performance using the interfacial energy concept. In addition, surface thermodynamics was applied to understand and predict surface sampling performance. Wipe materials were directly inoculated with known concentrations of B. anthracis spores and placed into extraction solutions, followed by sonication or vortexing. Experimental factors investigated included wipe material (polyester, cotton, and polyester-rayon), extraction solution (sterile deionized water [H(2)O], deionized water with 0.04% Tween 80 [H(2)O-T], phosphate-buffered saline [PBS], and PBS with 0.04% Tween 80 [PBST]), and physical dissociation method (vortexing or sonication). The most efficient extraction from wipes was observed for solutions containing the nonionic surfactant Tween 80. The increase in extraction efficiency due to surfactant addition was attributed to an attractive interfacial energy between Tween 80 and the centrifuge tube wall, which prevented spore adhesion. Extraction solution significantly impacted the extraction efficiency, as determined by statistical analysis (P < 0.05). Moreover, the extraction solution was the most important factor in extraction performance, followed by the wipe material. Polyester-rayon was the most efficient wipe material for releasing spores into solution by rank; however, no statistically significant difference between polyester-rayon and cotton was observed (P > 0.05). Vortexing provided higher spore recovery in H(2)O and H(2)O-T than sonication, when all three wipe materials and the reference control were considered (P < 0.05).

摘要

从常用拭子采样材料中提取炭疽杆菌孢子的效率的影响因素,并应用界面能概念来描述其性能。此外,还应用表面热力学来理解和预测表面采样性能。将拭子材料直接接种已知浓度的炭疽杆菌孢子,并放入提取溶液中,然后进行超声处理或涡旋。研究的实验因素包括拭子材料(聚酯、棉和涤棉)、提取溶液(无菌去离子水[H(2)O]、含 0.04%吐温 80 的去离子水[H(2)O-T]、磷酸盐缓冲盐水[PBS]和含 0.04%吐温 80 的 PBS[PBST])和物理分离方法(涡旋或超声处理)。从拭子中观察到最有效的提取是在含有非离子表面活性剂吐温 80 的溶液中。由于表面活性剂的添加而导致提取效率增加归因于吐温 80 和离心管壁之间的吸引力界面能,这防止了孢子的附着。提取溶液对提取效率有显著影响,这是通过统计分析确定的(P < 0.05)。此外,提取溶液是提取性能的最重要因素,其次是拭子材料。在释放孢子进入溶液方面,涤棉是最有效的拭子材料;然而,在聚酯和棉之间没有观察到统计学上的显著差异(P > 0.05)。当考虑所有三种拭子材料和参考对照时,涡旋在 H(2)O 和 H(2)O-T 中提供了比超声处理更高的孢子回收率(P < 0.05)。

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