Using digital droplet PCR to improve the detection of helminth infections of ruminants in Northern Ireland

使用数字液滴 PCR 改进北爱尔兰反刍动物蠕虫感染的检测

• 批准号: BB/V017276/1
• 负责人: Geoffrey Gobert
• 金额: $ 19.05万
• 依托单位: Queen's University Belfast
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV017276%2F1
• 关键词: Using; digital; droplet; PCR; improve

Fasciola hepatica, usually termed "the temperate liver fluke", is found worldwide and causes a disease known as fasciolosis. These helminth parasites is primarily a damaging infection of production ruminants. In Europe, the disease has been reported in UK, Ireland, France, Portugal, Spain, Switzerland, Italy, The Netherlands, Germany and Poland. As a food-borne disease, fasciolosis results from consumption of the infective larvae by ruminant animals. These helminths greatly reduce global agricultural capacity and outputs of both developing and developed communities, causing production inefficiency in millions of ruminants worldwide. Fasciolosis is also an important zoonosis, and is acknowledged by the World Health Organization (WHO).All organisms shed DNA into their environment, this material is known as eDNA. The detection of this eDNA enables the classification of species diversity and abundance in an environment. When Fasciola parasites are present, during part of their lifecycle they are exposed in the environment in order to be infectious to the snail or animal host, they also leave eDNA behind. The improved detection of this eDNA is the basis of this project. While low levels of this materials is expected in water or soil, or on plants, new molecular methods will be used to overcome these difficulties.These methods include the use of third generation technology droplet digital PCR (ddPCR) which greatly increases the sensitivity and reproducibility of DNA detection. ddPCR has the distinct value of being a fully quantitative method, and being highly sensitive and reproducible. Our proposal will drive a step-change in the way eDNA analysis is performed for the identification of helminth parasite infection risk. This will be involve examining a range of sample types, including water, soil, grass and faeces. These challenging samples will be characterised by the highly sensitive ddPCR technology to provide comprehensive quantitative measurements of parasite/vector eDNA on collected samples. Sheep farms in Northern Ireland will sampled over one year and these new sample collected to find molecular indicators of disease. This represents a less invasive and potentially less labor intensive approach to disease monitoring.In detecting parasitic infections earlier, the use of drugs (anthelmintics) can be reduced or avoided through intelligent pasture management - rotating animals around pastures according to their age, species and season, relative to the perception of infection risk in those pastures. Farmers would be able to make these decisions more effectively if they had access to relevant data from their own farm.

肝片形吸虫，通常被称为“温和的肝吸虫”，是世界各地发现，并导致一种疾病称为肝片形吸虫病。这些蠕虫寄生虫主要是生产反刍动物的破坏性感染。在欧洲，英国、爱尔兰、法国、葡萄牙、西班牙、瑞士、意大利、荷兰、德国和波兰均报告了该病。作为一种食源性疾病，片形吸虫病是由反刍动物食用感染性幼虫引起的。这些蠕虫极大地降低了发展中国家和发达国家的全球农业生产能力和产量，导致全世界数百万反刍动物的生产效率低下。片形吸虫病也是一种重要的人畜共患病，并被世界卫生组织（WHO）所承认。所有生物都会将DNA释放到其环境中，这种物质被称为eDNA。这种eDNA的检测使得能够对环境中的物种多样性和丰度进行分类。当片形吸虫寄生虫存在时，在其生命周期的一部分期间，它们暴露在环境中以感染蜗牛或动物宿主，它们也会留下eDNA。该eDNA的改进检测是该项目的基础。虽然预计这种物质在水、土壤或植物中的含量很低，但将使用新的分子方法来克服这些困难，这些方法包括使用第三代技术液滴数字PCR（ddPCR），该技术大大提高了DNA检测的灵敏度和重现性。ddPCR具有完全定量、灵敏度高、重复性好的独特价值。我们的提案将推动eDNA分析识别蠕虫寄生虫感染风险的方式发生重大变化。这将涉及检查一系列样品类型，包括水、土壤、草和粪便。这些具有挑战性的样本将通过高灵敏度的ddPCR技术进行表征，以提供对采集样本的寄生虫/载体eDNA的全面定量测量。北方爱尔兰的羊场将在一年内取样，收集这些新样品以寻找疾病的分子指标。这是一种侵入性更小、劳动强度更低的疾病监测方法。在早期检测寄生虫感染时，可以通过智能牧场管理减少或避免药物（驱虫剂）的使用-根据牧场感染风险的感知，根据年龄、物种和季节在牧场周围轮换动物。如果农民能够从自己的农场获得相关数据，他们将能够更有效地做出这些决定。