Comparison of four concentration methods of adenovirus, norovirus and rotavirus in tap water

IF 2.2 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS
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引用次数: 0

Abstract

Human enteric viruses, as adenovirus (HAdV), norovirus (HuNoV) and rotavirus (RVA) are significant causes of gastroenteritis associated with consumption of contaminated water worldwide. Various methods have been described for their detection and monitoring in water. The aim of this study was to compare the performance of four conditions for concentrating HAdV, HuNoV and RVA from water matrices, in order to develop a single protocol that could simultaneously concentrate all target viruses from tap water. The tested conditions were based on the adsorption-elution using electronegative filters, in which we evaluated cation-coated filtration by MgCl2 with or without acid rinse by H2SO4 and two elution buffers, namely NaOH and tris-glycine-beef extract. Genomic material was extracted and amplified by real-time PCR and real-time RT-PCR using commercial kits. Based on the statistical analysis of amplification results (cycles of quantification), the condition involving cation-coated filtration by MgCl2 using electronegative filters with acid rinse by H2SO4 combined with NaOH elution allowed efficient recovery of both HAdV, HuNoV and RVA from tap water compared to the other conditions. These findings confirm the effectiveness of the approach used to monitor three major enteric viruses in tap water.

自来水中腺病毒、诺如病毒和轮状病毒四种浓缩方法的比较
腺病毒 (HAdV)、诺如病毒 (HuNoV) 和轮状病毒 (RVA) 等人类肠道病毒是世界各地因饮用受污染的水而引发肠胃炎的主要原因。目前已有多种方法用于检测和监测水中的这些病毒。本研究的目的是比较从水基质中浓缩 HAdV、HuNoV 和 RVA 的四种条件的性能,以开发一种可同时从自来水中浓缩所有目标病毒的单一方案。测试条件基于使用电负性过滤器的吸附-洗脱法,其中我们评估了阳离子涂层过滤法,即使用 MgCl2、H2SO4 酸洗或不使用 H2SO4 酸洗,以及两种洗脱缓冲液(即 NaOH 和三甘氨酸-牛肉提取物)。提取基因组材料并使用商业试剂盒进行实时 PCR 和实时 RT-PCR 扩增。根据对扩增结果(定量周期)的统计分析,与其他条件相比,在阳离子涂层过滤条件下,使用电负性过滤器进行 MgCl2 过滤,并用 H2SO4 酸冲洗,再用 NaOH 洗脱,可从自来水中有效回收 HAdV、HuNoV 和 RVA。这些发现证实了用于监测自来水中三种主要肠道病毒的方法的有效性。
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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
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