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利用细胞培养-聚合酶链式反应(qPCR)集成检测方法快速定量中国渤海湾地表水中的肠道病毒。

Rapid quantification of infectious enterovirus from surface water in Bohai Bay, China using an integrated cell culture-qPCR assay.

机构信息

College of Environmental Science and Engineering, Key Laboratory of Pollution and Environmental Criteria, Ministry of Education, Nankai University, Tianjin 300071, China.

出版信息

Mar Pollut Bull. 2011 Oct;62(10):2047-54. doi: 10.1016/j.marpolbul.2011.07.024. Epub 2011 Sep 1.

Abstract

To rapidly quantify infectious enteroviruses polluting the coastal seawaters, a newly developed integrated cell culture and reverse transcription quantitative PCR (ICC-RT-qPCR) assay was used to identify the contamination by enteroviruses in winter seawater samples of Bohai Bay, Tianjin, China. The gene copies of enteroviral 5'UTRs correlated to the initial inoculum numbers across the concentration range of 0.05-500 PFU mL(-1) (correlation coefficient (R(2)) was 0.9667). ICC-qPCR revealed that five of seven samples (70.4%) were positive for infectious enteroviruses. The concentration of enteroviruses was estimated at 0.2-21 PFU L(-1). The result demonstrated that the contamination of enteroviruses in this coastal area may constitute a potential public health risk. This study established a practical assay for widespread monitoring studies of aquatic environments for viral contamination and provided meaningful data for human waterborne viral risk assessment.

摘要

为了快速定量检测污染沿海海水的肠道病毒,本研究采用了一种新建立的整合细胞培养和反转录定量聚合酶链反应(ICC-RT-qPCR)检测方法,对中国天津渤海湾冬季海水中肠道病毒的污染情况进行了鉴定。肠道病毒 5'UTR 的基因拷贝数与初始接种物数量在 0.05-500PFU/mL 的浓度范围内呈相关性(相关系数(R(2))为 0.9667)。ICC-qPCR 结果显示,7 个样本中有 5 个(70.4%)为肠道病毒阳性。肠道病毒的浓度估计为 0.2-21PFU/L。该结果表明,该沿海地区肠道病毒的污染可能构成潜在的公共卫生风险。本研究建立了一种实用的检测方法,可广泛用于监测水生环境中的病毒污染情况,并为人体经水传播病毒风险评估提供了有意义的数据。

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