首页> 外文期刊>Water Research >Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study
【24h】

Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

机译:在多实验室比较研究中犬相关细菌科检定的性能评估

获取原文
获取原文并翻译 | 示例
           

摘要

The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment.
机译:在城市地区,狗粪便污染的影响可能很大,而且经常被低估。利用定量PCR编码细菌的16S rRNA的狗相关基因序列的微生物来源跟踪方法(MST)是评估这些贡献的有用工具。然而,关于可用测定的性能的数据很少。此处提供了一项多实验室研究的结果,该结果测试了从加利福尼亚州收集的粪便样本中创建的64种单一和混合粪便来源样本中,两种测定狗相关拟杆菌(DogBact和BacCan-UCD)的测定方法。 qPCR数据处理的标准化降低了实验室间灵敏度和特异性结果的变异性。两种测定均显示出100%的灵敏度。提出了消除随机和确认的非目标响应的归一化方法。标准化qPCR数据处理,通过非靶标特异性拟杆菌鉴定(GenBac3)进行归一化以及使用阈值标准的结合显着提高了两种测定的计算特异性。这样的措施不仅将合理地改善MST数据的解释性,不仅适用于犬类相关分析,而且还适用于用于识别和监测环境中粪便污染的所有qPCR分析。

著录项

  • 来源
    《Water Research》 |2013年第18期|6909-6920|共12页
  • 作者单位

    Department of Civil and Environmental Engineering, University of California Davis, One Shields Ave, Davis, CA 95616, USA;

    National Oceanic and Atmospheric Administration, Atlantic Oceanographic and Meteorological Laboratory, Miami, FL, USA;

    National Oceanic and Atmospheric Administration, Atlantic Oceanographic and Meteorological Laboratory, Miami, FL, USA;

    National Oceanic and Atmospheric Administration, Northwest Fisheries Science Center Laboratory, Seattle, WA, USA;

    Cooperative Institute for Marine and Atmospheric Studies, University of Miami, Miami, FL, USA;

    Cooperative Institute for Marine and Atmospheric Studies, University of Miami, Miami, FL, USA;

    Department of Civil and Environmental Engineering, University of California Los Angeles, 5732 Boelter Hall, Los Angeles, CA, USA;

    Department of Civil and Environmental Engineering, University of California Davis, One Shields Ave, Davis, CA 95616, USA,Department of Civil and Environmental Engineering, University of California Los Angeles, 5732 Boelter Hall, Los Angeles, CA, USA;

    Bren School of Environmental Science & Management and Earth Research Institute, University of California, Santa Barbara, CA, USA;

    VCD School of Biomolecular and Biomedical Science, University College Dublin, Dublin 4, Ireland;

    U.S. Environmental Protection Agency, Office of Research and Development, National Risk Management Research Laboratory, Cincinnati, OH, USA;

    U.S. Environmental Protection Agency, Office of Research and Development, National Risk Management Research Laboratory, Cincinnati, OH, USA;

    VCD School of Biomolecular and Biomedical Science, University College Dublin, Dublin 4, Ireland;

    Southern California Coastal Water Research Project, Costa Mesa, CA, USA;

    U.S Environmental Protection Agency, Microbial Contaminants Control Branch, National Risk Management Research Laboratory, Cincinnati, OH, USA;

    Department of Civil and Environmental Engineering, University of California Los Angeles, 5732 Boelter Hall, Los Angeles, CA, USA;

    Bren School of Environmental Science & Management and Earth Research Institute, University of California, Santa Barbara, CA, USA;

    Department of Civil and Environmental Engineering, University of California Davis, One Shields Ave, Davis, CA 95616, USA,Singapore Centre on Environmental Life Sciences Engineering (SCELSE), School of Biological Sciences and School of Civil and Environmental Engineering, Nanyang Technological University, Singapore,Department of Civil and Environmental Engineering, University of California Davis, One Shields Ave, Davis, CA 95616, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Quantitative PCR; Microbial source tracking; Bacteroidales; Canine fecal pollution;

    机译:定量PCR;微生物来源跟踪;拟杆菌属;犬粪便污染;

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