便携式自动生物气溶胶采样系统,用于快速现场检测目标空气微生物。

Journal of Environmental Monitoring Pub Date : 2012-10-26 Epub Date: 2012-09-05 DOI:10.1039/c2em30317e
Evgeny V Usachev, Anna V Pankova, Elina A Rafailova, Oleg V Pyankov, Igor E Agranovski
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引用次数: 16

摘要

生物气溶胶可引起各种严重的人类和动物疾病,对其及时、定性、精确的检测和控制正成为一个重要的科技课题。近几十年来,生物气溶胶检测已成为生物防御领域的一个重要课题。已经开发了许多类型的便携式和固定式生物气溶胶采样器,并且在某些情况下,集成到利用各种微生物学技术对收集的微生物进行分析的自动检测系统中。本文介绍了一种个人采样器与便携式实时PCR技术结合使用。结果发现,一种荧光染料可以成功地以多种形式在一个PCR管中对许多目标生物气溶胶进行定性检测,使所建议的技术成为可靠的“第一警报”装置。这种方法已被专门开发并成功验证用于便携式PCR装置快速检测目标微生物,这在现场条件下尤其重要,其中感兴趣的微生物数量通常超过可用的PCR反应管的数量。该方法可以检测目标微生物,并触发一些相应的卫生检疫程序,以定位危险感染可能的传播。在受控的实验室条件下对样品进行详细分析后,可用于准确地确定目标组中哪些特定微生物已在现场快速检测到。个人采样器即使对小尺寸病毒(>20 nm)的采集效率也高于90%,并且在较长时间内性能稳定。此外,研究发现,对于本项目使用的微生物(噬菌体MS2和T4),在样品制备过程中省去核酸分离纯化步骤不会导致系统灵敏度降低,这对未来微型生物气溶胶监测仪器的开发具有极其重要的意义。
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Portable automatic bioaerosol sampling system for rapid on-site detection of targeted airborne microorganisms.

Bioaerosols could cause various severe human and animal diseases and their opportune and qualitative precise detection and control is becoming a significant scientific and technological topic for consideration. Over the last few decades bioaerosol detection has become an important bio-defense related issue. Many types of portable and stationary bioaerosol samplers have been developed and, in some cases, integrated into automated detection systems utilizing various microbiological techniques for analysis of collected microbes. This paper describes a personal sampler used in conjunction with a portable real-time PCR technique. It was found that a single fluorescent dye could be successfully used in multiplex format for qualitative detection of numerous targeted bioaerosols in one PCR tube making the suggested technology a reliable "first alert" device. This approach has been specifically developed and successfully verified for rapid detection of targeted microorganisms by portable PCR devices, which is especially important under field conditions, where the number of microorganisms of interest usually exceeds the number of available PCR reaction tubes. The approach allows detecting targeted microorganisms and triggering some corresponding sanitary and quarantine procedures to localize possible spread of dangerous infections. Following detailed analysis of the sample under controlled laboratory conditions could be used to exactly identify which particular microorganism out of a targeted group has been rapidly detected in the field. It was also found that the personal sampler has a collection efficiency higher than 90% even for small-sized viruses (>20 nm) and stable performance over extended operating periods. In addition, it was found that for microorganisms used in this project (bacteriophages MS2 and T4) elimination of nucleic acids isolation and purification steps during sample preparation does not lead to the system sensitivity reduction, which is extremely important for development of miniature bioaerosol monitoring instrumentation in the future.

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来源期刊
Journal of Environmental Monitoring
Journal of Environmental Monitoring 环境科学-分析化学
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