利用纳米氧化铝纤维滤器浓缩环境水样中的病毒。
Concentration of viruses from environmental waters using nanoalumina fiber filters.
机构信息
Civil and Environmental Engineering, University of California, Irvine, CA 92697, USA.
出版信息
J Microbiol Methods. 2010 Apr;81(1):33-8. doi: 10.1016/j.mimet.2010.01.018. Epub 2010 Jan 28.
Human viral contamination in drinking and recreational water may persist for extensive periods of time and cause a significant health risk concern. The aim of this study is to evaluate a viral recovery method using a new electropositive charged nanoalumina filter and to compare results with the widely used negatively charged HAWP filter by Millipore Inc. The recovery of infectious recombinant adenovirus type 5 (rAd5) was tested using the Fluorescence-Activated Cell Sorting (FACS) assay, in parallel with viral genomes recovery assay by quantitative PCR (qPCR). The mean infectivity recoveries were 82-91% by nanoalumina filters eluted with 3% beef extract (BE, pH 6.0), and 78-90% by HAWP filters eluted with 3% BE (pH 9.0), respectively, from 1L of environmental samples seeded with 1 pfu/mL rAd5. The mean genome recoveries were 16-35% by nanoalumina filters eluted with BE (pH 6.0), and 29-66% by HAWP filters eluted with NaOH (pH 10.8) from different types of water, respectively. Water quality, concentration of viruses, filters, and elution buffers are factors that determine the viral recovery efficiencies. The nanoalumina filters also had higher filtration rates than HAWP filters for large volumes of environmental water samples (up to 10 L), thus, have an advantage in concentrating infectious viruses from environments without pre-filtration, adjusting pH or adding multivalent cations.
人类病毒污染在饮用水和娱乐水中可能会持续很长时间，并造成重大的健康风险。本研究旨在评估一种使用新型正电荷纳米氧化铝过滤器的病毒回收方法，并与密理博公司广泛使用的负电荷 HAWP 过滤器进行比较。采用荧光激活细胞分选（FACS）检测法和定量 PCR（qPCR）检测病毒基因组回收法，对传染性重组腺病毒 5 型（rAd5）的回收进行了测试。用 3%牛肉提取物（BE，pH6.0）洗脱纳米氧化铝过滤器，平均感染性回收率为 82-91%，用 3%BE（pH9.0）洗脱 HAWP 过滤器，平均回收率为 78-90%，从 1L 接种 1 pfu/mL rAd5 的环境样本中回收。用 BE（pH6.0）洗脱纳米氧化铝过滤器，平均基因组回收率为 16-35%，用 NaOH（pH10.8）洗脱 HAWP 过滤器，平均回收率为 29-66%，分别从不同类型的水中回收。水质、病毒浓度、过滤器和洗脱缓冲液是决定病毒回收率的因素。纳米氧化铝过滤器对于大体积的环境水样（高达 10L）的过滤速度也高于 HAWP 过滤器，因此，在不进行预过滤、调整 pH 值或添加多价阳离子的情况下，从环境中浓缩传染性病毒具有优势。
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