Slana Iva, Bier Nadja, Bartosova Barbora, Marucci Gianluca, Possenti Alessia, Mayer-Scholl Anne, Jokelainen Pikka, Lalle Marco
Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296/70, 621 00 Brno, Czech Republic.
Department of Food Safety, German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Str 8-10, 10589 Berlin, Germany.
Microorganisms. 2021 Jan 13;9(1):167. doi: 10.3390/microorganisms9010167.
Human infection with the important zoonotic foodborne pathogen has been associated with unwashed raw fresh produce consumption. The lack of a standardised detection method limits the estimation of fresh produce as an infection source. To support method development and standardisation, an extensive literature review and a multi-attribute assessment were performed to analyse the key aspects of published methods for the detection of oocyst contamination in fresh produce. Seventy-seven published studies were included, with 14 focusing on fresh produce. Information gathered from expert laboratories via an online questionnaire were also included. Our findings show that procedures for oocyst recovery from fresh produce mostly involved sample washing and pelleting of the washing eluate by centrifugation, although washing procedures and buffers varied. DNA extraction procedures including mechanical or thermal shocks were identified as necessary steps to break the robust oocyst wall. The most suitable DNA detection protocols rely on qPCR, mostly targeting the gene or the bp repetitive element. When reported, validation data for the different detection methods were not comparable and none of the methods were supported by an interlaboratory comparative study. The results of this review will pave the way for an ongoing development of a widely applicable standard operating procedure.
人类感染这种重要的人畜共患食源性病原体与食用未清洗的生鲜农产品有关。缺乏标准化的检测方法限制了将生鲜农产品作为感染源的评估。为支持方法的开发和标准化，我们进行了广泛的文献综述和多属性评估，以分析已发表的生鲜农产品中卵囊污染检测方法的关键方面。纳入了77项已发表的研究，其中14项聚焦于生鲜农产品。还纳入了通过在线问卷从专业实验室收集的信息。我们的研究结果表明，从生鲜农产品中回收卵囊的程序大多包括样品清洗以及通过离心对清洗洗脱液进行沉淀，不过清洗程序和缓冲液有所不同。包括机械或热冲击的DNA提取程序被确定为打破坚固卵囊壁的必要步骤。最合适的DNA检测方案依赖于定量聚合酶链反应（qPCR），大多靶向基因或bp重复元件。当报告不同检测方法的验证数据时，这些数据不可比，且没有一种方法得到实验室间比较研究的支持。本综述的结果将为正在进行的广泛适用的标准操作规程的制定铺平道路。
