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Methods to detect infectious human enteric viruses in environmental water samples

机译:检测环境水样中传染性人类肠道病毒的方法

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摘要

Currently, a wide range of analytical methods is available for virus detection in environmental water samples. Molecular methods such as polymerase chain reaction (PCR) and quantitative real time PCR (qPCR) have the highest sensitivity and specificity to investigate virus contamination in water, so they are the most commonly used in environmental virology. Despite great sensitivity of PCR, the main limitation is the lack of the correlation between the detected viral genome and viral infectivity, which limits conclusions regarding the significance for public health. To provide information about the infectivity of the detected viruses, cultivation on animal cell culture is the gold standard. However, cell culture infectivity assays are laborious, time consuming and costly. Also, not all viruses are able to produce cytopathic effect and viruses such as human noroviruses have no available cell line for propagation. In this brief review, we present a summary and critical evaluation of different approaches that have been recently proposed to overcome limitations of the traditional cell culture assay and PCR assay such as integrated cell culture-PCR, detection of genome integrity, detection of capsid integrity, and measurement of oxidative damages on viral capsid protein. Techniques for rapid detection of infectious viruses such as fluorescence microscopy and automated flow cytometry have also been suggested to assess virus infectivity in water samples.
机译:当前,有多种分析方法可用于检测环境水样中的病毒。诸如聚合酶链反应(PCR)和定量实时PCR(qPCR)之类的分子方法对研究水中的病毒污染具有最高的灵敏度和特异性,因此它们是环境病毒学中最常用的方法。尽管PCR具有很高的灵敏度,但主要的局限性在于检测到的病毒基因组与病毒感染性之间缺乏相关性,这限制了有关对公共卫生重要性的结论。为了提供有关检测到的病毒的感染性的信息,在动物细胞培养上进行培养是金标准。然而,细胞培养物感染性测定费力,费时且昂贵。而且,并非所有病毒都能产生细胞病变作用,并且诸如人诺如病毒的病毒也没有可用的细胞系用于繁殖。在这篇简短的评论中,我们将对最近提出的各种方法进行总结和批判性评估,以克服传统细胞培养测定法和PCR测定法的局限性,例如集成细胞培养PCR,基因组完整性检测,衣壳完整性检测,和测量病毒衣壳蛋白的氧化损伤。还提出了快速检测传染性病毒的技术,例如荧光显微镜和自动流式细胞术,以评估水样中的病毒感染性。

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