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Parameters Affecting Spore Recovery from Wipes Used in Biological Surface Sampling

机译:影响从生物表面采样中使用的抹布回收孢子的参数

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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).
机译:对精确和可靠地收集潜在生物威胁污染物的需求已激发了研究,以加深对生物表面采样方法变异性的了解。在这种情况下,这项工作的目的是确定影响从常用擦拭采样材料中提取炭疽芽孢杆菌斯特恩孢子效率的参数,并使用界面能概念来描述性能。另外,应用表面热力学来理解和预测表面采样性能。将擦拭材料直接接种已知浓度的炭疽芽孢杆菌孢子,并将其置于提取液中,然后进行超声处理或涡旋。研究的实验因素包括擦拭材料(聚酯,棉和聚酯人造丝),萃取液(无菌去离子水[H_(2)O],含0.04%Tween 80的去离子水[H_(2)OT],磷酸盐缓冲盐水) [PBS],以及含0.04%Tween 80的PBS [PBST])和物理解离方法(涡旋或超声处理)。对于包含非离子表面活性剂吐温80的溶液,观察到了从擦拭布中最有效的提取。由于添加表面活性剂而导致的提取效率提高归因于吐温80与离心管壁之间的界面能极强,从而阻止了孢子粘附。通过统计分析确定,提取液显着影响提取效率(P <0.05)。此外,萃取液是萃取性能中最重要的因素,其次是擦拭材料。聚酯人造丝是最有效的将孢子按等级释放到溶液中的擦拭材料。然而,在涤纶人造丝和棉花之间没有观察到统计学上的显着差异(P> 0.05)。当考虑所有三种擦拭材料和参考对照时,涡旋作用在H_(2)O和H_(2)O-T中提供的孢子回收率高于超声处理(P <0.05)。

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