评估从盐水中浓缩各种病毒的自动超滤系统。

IF 4.1 2区 农林科学 Q2 ENVIRONMENTAL SCIENCES Food and Environmental Virology Pub Date : 2024-06-29 DOI:10.1007/s12560-024-09602-6
Simran Singh, Tiong Gim Aw, Joan B. Rose
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引用次数: 0

摘要

环境水体中的致病病毒含量通常太低,无法直接检测,因此通常需要浓缩步骤来提高分析灵敏度。本研究的目的是评估一种自动过滤装置 Innovaprep 浓缩移液管选择(CP Select),用于快速浓缩盐水样本中的病毒,同时考虑处理过程的持续时间和使用的方便性。将四种噬菌体(MS2、P22、Phi6 和 PhiX174)和三种动物病毒(腺病毒、冠状病毒 OC43 和犬瘟热病毒)播种到人工海水、水族馆水和海湾水样本中,并使用 CP Select 进行处理。病毒的回收率是通过斑块检测法或液滴数字 PCR(ddPCR)测定的。使用斑块检测法,噬菌体的平均回收率从 4.84 ± 3.8% 到 82.73 ± 27.3% 不等,其中 P22 噬菌体的回收率最高。经 ddPCR 测定,CP Select 的平均回收率分别为腺病毒(39.31 ± 26.6%)、冠状病毒 OC43(19.04 ± 11.6%)和犬瘟热病毒(19.84 ± 13.6%)。总体而言,病毒基因组组成而非病毒大小影响了 CP Select 的回收率。该系统的超滤移液器使用的样品量较小,这妨碍了将该方法用作海水中病毒的主要浓缩步骤。不过,当需要快速检测高度污染水体(如废水或被污水污染的地表水)中的病毒时,CP Select 的易用性和快速处理时间尤为有利。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Evaluation of an Automated Ultrafiltration System for Concentrating a Range of Viruses from Saline Waters

Pathogenic viruses in environmental water are usually present in levels too low for direct detection and thus, a concentration step is often required to increase the analytical sensitivity. The objective of this study was to evaluate an automated filtration device, the Innovaprep Concentrating Pipette Select (CP Select) for the rapid concentration of viruses in saline water samples, while considering duration of process and ease of use. Four bacteriophages (MS2, P22, Phi6, and PhiX174) and three animal viruses (adenovirus, coronavirus OC43, and canine distemper virus) were seeded in artificial seawater, aquarium water, and bay water samples, and processed using the CP Select. The recovery efficiencies of viruses were determined either using a plaque assay or droplet digital PCR (ddPCR). Using plaque assays, the average recovery efficiencies for bacteriophages ranged from 4.84 ± 3.8% to 82.73 ± 27.3%, with highest recovery for P22 phage. The average recovery efficiencies for the CP Select were 39.31 ± 26.6% for adenovirus, 19.04 ± 11.6% for coronavirus OC43, and 19.84 ± 13.6% for canine distemper virus, as determined by ddPCR. Overall, viral genome composition, not the size of the virus, affected the recovery efficiencies for the CP Select. The small sample volume size used for the ultrafilter pipette of the system hinders the use of this method as a primary concentration step for viruses in marine waters. However, the ease of use and rapid processing time of the CP Select are especially beneficial when rapid detection of viruses in highly contaminated water, such as wastewater or sewage-polluted surface water, is needed.

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来源期刊
Food and Environmental Virology
Food and Environmental Virology ENVIRONMENTAL SCIENCES-MICROBIOLOGY
CiteScore
6.50
自引率
2.90%
发文量
35
审稿时长
1 months
期刊介绍: Food and Environmental Virology publishes original articles, notes and review articles on any aspect relating to the transmission of pathogenic viruses via the environment (water, air, soil etc.) and foods. This includes epidemiological studies, identification of novel or emerging pathogens, methods of analysis or characterisation, studies on survival and elimination, and development of procedural controls for industrial processes, e.g. HACCP plans. The journal will cover all aspects of this important area, and encompass studies on any human, animal, and plant pathogenic virus which is capable of transmission via the environment or food.
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