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Portable automatic bioaerosol sampling system for rapid on-site detection of targeted airborne microorganisms

机译: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中的微生物

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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.
机译:生物气溶胶可引起各种严重的人类和动物疾病,其适当和定性的精确检测和控制正成为重要的科学技术课题。在过去的几十年中,生物气溶胶检测已经成为与生物防御相关的重要问题。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,并在某些情况下将其集成到利用各种微生物技术来分析收集的微生物的自动检测系统中。本文介绍了与便携式实时PCR技术结合使用的个人采样器。已经发现,一种荧光染料可以成功地以多重形式用于定性检测一个PCR管中的许多目标生物气溶胶,从而使该技术成为可靠的“第一警报”设备。该方法已经过专门开发并成功验证,可通过便携式PCR装置快速检测目标微生物,这在现场条件下尤其重要,在这种情况下,目标微生物的数量通常会超过可用PCR反应管的数量。该方法允许检测目标微生物并触发一些相应的卫生和检疫程序以定位危险感染的可能传播。在可控的实验室条件下对样品进行详细分析之后,可以用来准确识别目标群体中哪些特定微生物已在野外快速检测到。还发现,即使对于小型病毒(> 20 nm),个人采样器也具有高于90%的采集效率,并且在较长的操作时间内仍具有稳定的性能。此外,还发现对于该项目中使用的微生物(噬菌体MS2和T4),在样品制备过程中消除核酸分离和纯化步骤不会导致系统灵敏度降低,这对于开发微型生物气溶胶监测仪器至关重要在将来。

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