Evaluation of Filters for the Sampling and Quantification of RNA Phage Aerosols
Evaluation of Filters for the Sampling and Quantification of RNA Phage Aerosols
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DOI：
10.1080/02786826.2010.501351
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发表时间：
2010-01-01
影响因子：
5.2
通讯作者：
Duchaine, Caroline
中科院分区：
文献类型：
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作者：
Gendron, Louis;Verreault, Daniel;Duchaine, Caroline
这项研究利用了有毒的噬菌体phi 6（dsRNA）和MS 2（ssRNA）作为空气中的RNA病毒的替代品。两种不同的过滤器类型，聚四氟乙烯（PTFE）和聚碳酸酯（PC），测试其收集雾化RNA的效率。测试了两种商业试剂盒的总RNA分离。此外，在三种不同的培养基中进行热休克处理，以获得用于dsRNA的逆转录测定的最有利条件。我们的研究结果表明，PC过滤器更适合于回收由空斑试验确定的感染性空气中的RNA病毒。两种类型的过滤器在从气溶胶化的噬菌体phi 6中回收RNA方面同样有效，如通过qRT-PCR建立的。病毒样品应使用QIAamp Viral RNA Mini Kit处理，并在RT-PCR前在TE缓冲液中于110 ℃下热休克处理5分钟，以最大限度地检测噬菌体phi 6。总体而言，回收的病毒的感染性受到雾化/空气采样过程的严重影响，应通过qRT-PCR确定空气样本中是否存在RNA病毒。
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 degrees 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.