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实时 PCR 检测日本河水中的腺病毒、多瘤病毒和扭结藤病毒。

Real-time PCR detection of adenoviruses, polyomaviruses, and torque teno viruses in river water in Japan.

机构信息

Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 4-3-11 Takeda, Kofu, Yamanashi 400-8511, Japan.

出版信息

Water Res. 2010 Mar;44(6):1747-52. doi: 10.1016/j.watres.2009.11.043. Epub 2009 Nov 27.

Abstract

The prevalence of DNA viruses in water from the Tamagawa River, Japan was quantitatively surveyed for 6months, from April to September 2003. A total of 18 river water samples were subjected to virus concentration method using an electronegative membrane, followed by DNA extraction and direct quantitative real-time polymerase chain reaction (qPCR) for DNA viruses. Adenoviruses of serotypes 40 and 41 were detected most frequently in the river water samples tested (61.1%), at a concentration ranging from 3.16x10(3) to 1.38x10(5) copies/l, followed by JC polyomaviruses (11.1%) and torque teno viruses (5.6%). No sample was positive for BK polyomaviruses. In addition, for selective detection of virus particles, adenoviruses 40 and 41 were tested with qPCR combined with an immunomagnetic separation technique; they were detected in only 16.7% of the samples, showing a concentration ranging from 7.42x10(2) to 4.24x10(4) copies/l. This study is significant since it is the first study to demonstrate the prevalence of polyomaviruses in water samples in Japan and to use immunomagnetic separation qPCR to detect adenovirus particles in aquatic environments.

摘要

2003 年 4 月至 9 月,对日本玉川河水体中的 DNA 病毒进行了为期 6 个月的定量调查。共采集了 18 个河水样本,采用带负电荷的膜对病毒进行浓缩,然后提取 DNA,直接进行实时定量聚合酶链反应(qPCR)检测 DNA 病毒。在检测的河水样本中,血清型 40 和 41 的腺病毒最为常见(61.1%),浓度范围为 3.16x10(3)到 1.38x10(5)拷贝/升,其次是 JC 多瘤病毒(11.1%)和扭转型肠病毒(5.6%)。没有样本对 BK 多瘤病毒呈阳性。此外,为了选择性检测病毒颗粒,用 qPCR 结合免疫磁分离技术对腺病毒 40 和 41 进行了检测;仅在 16.7%的样本中检测到它们,浓度范围为 7.42x10(2)到 4.24x10(4)拷贝/升。本研究具有重要意义,因为它是首次在日本的水样中检测到多瘤病毒的流行,并使用免疫磁分离 qPCR 检测水生环境中的腺病毒颗粒。

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