...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Environmental Science & Technology >Smartphone-Based in-Gel Loop-Mediated Isothermal Amplification (gLAMP) System Enables Rapid Coliphage MS2 Quantification in Environmental Waters
【24h】

Smartphone-Based in-Gel Loop-Mediated Isothermal Amplification (gLAMP) System Enables Rapid Coliphage MS2 Quantification in Environmental Waters

机译：基于智能手机的凝胶内环介导的等温扩增（gLAMP）系统可在环境水域中对Collhage MS2快速定量

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Model coliphages (e.g., ΦX174, MS2, and PRD1) have been widely used as surrogates to study the fate and transport of pathogenic viruses in the environment and during wastewater treatment. Two groups of coliphages (F-specific and somatic) are being explored as indicators of viral fecal pollution in ambient water. However, the detection and quantification of coliphages still largely rely on time-consuming culture-based plaque assays. In this study, we developed an in-gel loop-mediated isothermal amplification (gLAMP) system enabling coliphage MS2 quantification within 30 min using standard laboratory devices. Viral particles (MS2) were immobilized with LAMP reagents in polyethylene glycol hydrogel, and then viral RNAs were amplified through a LAMP reaction. Due to the restriction effect of the hydrogel matrix, one viral particle would only produce one amplicon dot. Therefore, the sample virus concentrations can be determined based on the number of fluorescent amplicon dots using a smartphone for imaging. The method was validated by using artificially spiked and naturally contaminated water samples. gLAMP results were shown to correlate well with plaque assay counts (R ~(2) = 0.984, p < 0.05) and achieved similar sensitivity to quantitative reverse-transcription polymerase chain reaction (RT-qPCR; 1 plaque-forming unit per reaction). Moreover, gLAMP demonstrated a high level of tolerance against inhibitors naturally present in wastewater, in which RT-qPCR was completely inhibited. Besides MS2, gLAMP can also be used for the quantification of other microbial targets (e.g., Escherichia coli and Salmonella ). Considering its simplicity, sensitivity, rapidity, and versatility, gLAMP holds great potential for microbial water-quality analysis, especially in resource-limited settings.
机译：模型噬菌体（例如ΦX174，MS2和PRD1）已被广泛用作替代品，以研究环境和废水处理过程中致病病毒的命运和运输。目前正在研究两组大肠菌（F特异性和体细胞）作为环境水中病毒性粪便污染的指标。然而，对噬菌体的检测和定量仍主要依赖于耗时的基于培养物的噬菌斑测定。在这项研究中，我们开发了凝胶内环介导的等温扩增（gLAMP）系统，可使用标准实验室设备在30分钟内对大肠杆菌噬菌体MS2进行定量。将病毒颗粒（MS2）用LAMP试剂固定在聚乙二醇水凝胶中，然后通过LAMP反应扩增病毒RNA。由于水凝胶基质的限制作用，一个病毒颗粒只会产生一个扩增子点。因此，可以使用智能手机进行成像，根据荧光扩增子点的数量确定样本病毒的浓度。该方法通过使用人工加标和自然污染的水样品进行了验证。已显示gLAMP结果与噬斑测定计数密切相关（R〜（2）= 0.984，p <0.05），并且对定量逆转录聚合酶链反应（RT-qPCR; 1个噬斑每个反应形成单位）。此外，gLAMP表现出对废水中天然存在的抑制剂的高度耐受性，其中完全抑制了RT-qPCR。除MS2外，gLAMP还可以用于定量其他微生物靶标（例如大肠埃希氏菌和沙门氏菌）。考虑到其简单性，灵敏性，快速性和多功能性，gLAMP在微生物水质分析方面具有巨大潜力，尤其是在资源有限的环境中。

著录项

  • 来源
    《Environmental Science & Technology》 |2018年第11期|6399-6407|共9页
  • 作者

    Xiao Huang; Xingyu Lin; Katharina Urmann; Lijie Li; Xing Xie; Sunny Jiang; Michael R. Hoffmann;

  • 作者单位

    Linde + Robinson Laboratories, California Institute of Technology, Pasadena, California 91125, United States;

    Linde + Robinson Laboratories, California Institute of Technology, Pasadena, California 91125, United States;

    Linde + Robinson Laboratories, California Institute of Technology, Pasadena, California 91125, United States;

    Linde + Robinson Laboratories, California Institute of Technology, Pasadena, California 91125, United States;

    Linde + Robinson Laboratories, California Institute of Technology, Pasadena, California 91125, United States,School of Civil and Environmental Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, United States;

    Department of Civil and Environmental Engineering, Henry Samueli School of Engineering, University of California, Irvine, California 92697, United States;

    Linde + Robinson Laboratories, California Institute of Technology, Pasadena, California 91125, United States;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号