Comprehensive next-generation sequencing panel for foodborne pathogen detection and meat speciation

用于食源性病原体检测和肉类物种形成的综合下一代测序面板

基本信息

  • 批准号:
    10012313
  • 负责人:
  • 金额:
    $ 1.5万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2019
  • 资助国家:
    美国
  • 起止时间:
    2019-09-15 至 2021-06-30
  • 项目状态:
    已结题

项目摘要

Project Summary The Food and Drug Administration (FDA) regulates pet food, requiring it is safe to eat, contains no harmful substances, is produced using sanitary conditions, and is truthfully labeled. However, the foods do not need pre-market approval because they are made with ingredients considered safe. Pet food adulteration has been shown to occur in plants during processing, resulting in illness in animals and humans related to foodborne pathogens. Plant processing may also result in cross-contamination between products used for pet food production. This can result in pet foods having unintended ingredients, leading to false labelling. Whether intentional or not, adulteration of pet food with DNA from unlabeled genera has been previously detected. Current methods used to detect foodborne pathogens are labor-intensive and may take several days to obtain actionable results. Additionally, samples likely have to been sent to several different labs for comprehensive testing (e.g. for virus, bacteria, and parasites) and for detection of the genus of the meat source in the food. Additional testing capabilities, including comprehensive methods for rapid detection and initial characterization, would be beneficial for rapid recall of adulterated food. We propose to develop a targeted next-generation sequencing (NGS) panel, which incorporates primers for multiple pathogens associated with foodborne illness, for detection and initial characterization of the involved pathogen(s), and for characterization of the meat included in the product. Primers can be incorporated into the NGS workflow for multiplex PCR prior to sequencing. Use of primers allows for standardization of the NGS protocol for consistent performance across multiple labs and eases the use and implementation of the technology by those already familiar with PCR and Sanger sequencing. This renders implementation of NGS relatively simple in the diagnostic lab setting, even though NGS bioinformatics pipelines still need further development and validation for standardization across labs. The simplistic analysis of the data generated from a targeted NGS panel mitigates this limitation. Additionally, as a result of the initial PCR amplification steps, the test can be performed directly from the clinical sample or following an enrichment process. All of these advantages suggest that targeted NGS can be used to provide a comprehensive diagnostic assay for the detection of foodborne pathogens with initial pathogen characterization and detection of meat source genera, that has a more rapid turn-around time than currently used methods.
项目概要 美国食品和药物管理局 (FDA) 监管宠物食品,要求其可安全食用、含有 不含有害物质,在卫生条件下生产,并如实标识。 然而,这些食品不需要上市前批准,因为它们是用成分制成的 被认为是安全的。宠物食品掺假已被证明在加工过程中发生在植物中, 导致动物和人类罹患与食源性病原体相关的疾病。植物加工 还可能导致用于宠物食品生产的产品之间的交叉污染。这个可以 导致宠物食品含有非预期成分,从而导致错误标签。是否有意 不管是否如此,之前已经检测到宠物食品中掺有未标记属的 DNA。 目前用于检测食源性病原体的方法是劳动密集型的,可能需要花费数次时间 获得可行结果的天数。此外,样品可能已被发送到几个 不同的实验室进行综合测试(例如病毒、细菌和寄生虫)和检测 食品中肉类来源的属。额外的测试功能,包括 快速检测和初步表征的综合方法将有利于 快速召回掺假食品。 我们建议开发一个有针对性的下一代测序(NGS)面板,其中包含 与食源性疾病相关的多种病原体的引物,用于检测和初始 所涉及病原体的特征,以及包含在肉类中的特征 产品。在进行多重 PCR 之前,可以将引物纳入 NGS 工作流程中 测序。使用引物可以实现 NGS 协议的标准化,以实现一致 跨多个实验室的性能,并通过以下方式简化技术的使用和实施 那些已经熟悉 PCR 和桑格测序的人。这使得NGS的实施 尽管NGS生物信息学管道仍然存在,但在诊断实验室环境中相对简单 需要进一步开发和验证跨实验室的标准化。简单分析 从目标 NGS 面板生成的数据可以缓解这一限制。此外,作为 初始 PCR 扩增步骤的结果,可以直接从临床进行测试 样品或遵循浓缩过程。所有这些优点表明靶向 NGS 可用于为检测食源性细菌提供全面的诊断测定 具有初步病原体特征和肉源属检测的病原体, 比目前使用的方法更快的周转时间。

项目成果

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Rebecca Penrose Wilkes其他文献

Rebecca Penrose Wilkes的其他文献

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{{ truncateString('Rebecca Penrose Wilkes', 18)}}的其他基金

'COVID-19' Incorporation of SARS-CoV-2 primers into a targeted NGS panel for surveillance use in cats
“COVID-19”将 SARS-CoV-2 引物纳入目标 NGS 组合中,用于猫的监测
  • 批准号:
    10447438
  • 财政年份:
    2021
  • 资助金额:
    $ 1.5万
  • 项目类别:
TVDIL Infrastructure for CVM VET-LRN Veterinary Diagnostic Laboratory Program
CVM VET-LRN 兽医诊断实验室计划的 TVDIL 基础设施
  • 批准号:
    9474335
  • 财政年份:
    2017
  • 资助金额:
    $ 1.5万
  • 项目类别:
TVDIL Infrastructure for CVM VET-LRN Veterinary Diagnostic Laboratory Program
CVM VET-LRN 兽医诊断实验室计划的 TVDIL 基础设施
  • 批准号:
    9126296
  • 财政年份:
    2012
  • 资助金额:
    $ 1.5万
  • 项目类别:

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