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Evaluation of Filters for the Sampling and Quantification of RNA Phage Aerosols

机译:RNA噬菌体气溶胶采样和定量过滤器的评估

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摘要

This study exploits the virulent bacteriophages phi 6 (dsRNA) and MS2 (ssRNA) as surrogates for airborne RNA viruses. Two different filter types, polytetrafluoroethylene (PTFE) and polycarbonate (PC), were tested for their efficiency in collecting aerosolized RNA phages. Two commercial kits were tested for total RNA isolation. Also, heat shock treatments were performed in three different media to obtain the most favorable conditions for reverse transcription assays of dsRNA. Our findings suggest that PC filters are more suitable to recover infectious airborne RNA viruses as determined by plaque assays. Both types of filters were equally efficient in recovering RNA from aerosolized phage phi 6 as established by qRT-PCR. Viral samples should be treated with QIAamp Viral RNA Mini Kit and a 5 min heat shock treatment at 110°C in TE buffer before RT-PCR to maximize detection of phage phi 6. Overall, the infectivity of the recovered phages was severely affected by the aerosolization/air sampling process and the presence of RNA viruses in air samples should be determined by qRT-PCR.
机译:这项研究利用有毒的噬菌体phi 6(dsRNA)和MS2(ssRNA)作为空中RNA病毒的替代物。测试了两种不同的过滤器类型,聚四氟乙烯(PTFE)和聚碳酸酯(PC)收集雾化RNA噬菌体的效率。测试了两种商业试剂盒的总RNA分离。另外,在三种不同的介质中进行了热激处理,以获得用于dsRNA逆转录测定的最有利条件。我们的研究结果表明,通过噬菌斑测定,PC过滤器更适合回收传染性空气传播的RNA病毒。如通过qRT-PCR建立的,两种类型的过滤器在从雾化的噬菌体phi 6中回收RNA的效率均相同。病毒样品应使用QIAamp Viral RNA Mini Kit进行处理,并在RT-PCR之前在TE缓冲液中于110°C进行5分钟的热休克处理,以最大程度地检测噬菌体phi6。总体而言,回收的噬菌体的感染力受到气雾化/空气采样过程以及空气样品中是否存在RNA病毒应通过qRT-PCR确定。

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  • 来源
    《Aerosol Science and Technology》 |2010年第10期|p.893-901|共9页
  • 作者

    Louis Gendron;

  • 作者单位

    Centre de Recherche, Hôpital Laval, Institut Universitaire de Cardiologie et de Pneumologie de l’Université Laval, Quebec City, Quebec, Canada Département de Biochimie, Microbiologie et Bioinformatique, Faculté des Sciences et de Génie, UniversitÃ;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
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  • 入库时间 2022-08-18 00:57:42

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