Development of a Molecular Screening Tool for Detection and Identification of Cryptosporidium for Water Treatment Applications

隐孢子虫是一种存在于许多家畜和野生动物体内的病原体。它可能通过饮用水和娱乐用水系统传播给人类，导致严重的长期胃肠道感染，甚至死亡。这种病原体有许多种，其中只有3或4种已知对人类具有传染性。由于许多供水设施的农村性质以及相关的相对高降雨量，英国的此类病例相对较多。水处理设施使用显微镜常规监测隐孢子虫卵囊样结构。一旦确定了阳性样本，就需要在斯旺西使用DNA测序技术进行进一步分析，以确定确切的物种，并确定它是否会感染人类。在常见的混合污染中，鉴定仍然很困难。这不仅需要额外的成本和时间，还会给供水公司带来压力，因为处理设施可能会关闭，以确保在此期间不会有市民生病。这里提出了一个项目，开发一种分子工具，可以结合现有方法的两个元素。该工具将具有高通量，使许多样品能够并行处理，并且能够在竞争性的时间尺度内准确识别任何感染菌株。提出的主要目标是：1。开发和优化高通量分子工具，能够有效分离和鉴定特定种类的隐孢子虫，以单一和混合污染的形式存在；用实际样品的等分对分子工具进行验证，并将分子方法得到的结果与现有方法得到的结果进行比较。分子工具将实现高通量常规筛查能力，从而对潜在的疫情作出快速反应。除了降低可能关闭后的成本和声誉影响外，还为水务行业节省了分析时间。

Cryptosporidium is a pathogen that is found in a number of livestock and wild animals. It maybe passed to humans through drinking and recreational water systems and causes severeprolonged gastrointestinal infection, even death. There are numerous species of the pathogen,of which just 3 or 4 are known to be infectious to humans. The UK has a relatively highnumber of instances due to the rural nature of many water facilities and the associatedrelatively high rainfall.Water treatment facilities routinely monitor for Cryptosporidium oocyst-like structures usingmicroscopy. Once a positive sample is identified, further analysis using DNA sequencingtechniques at Swansea is necessary, to identify the exact species and ascertain whether it isinfective to humans. Identification remains difficult in mixed contaminations, which arecommon. This incurs extra cost, time and puts pressure on the water supplier through possibleshut down of treatment facilities to ensure no member of the public falls ill in the meantime.A project is proposed here, to develop a molecular tool that can combine the two elements ofthe existing methodology. The tool would be high throughput, enabling many samples to beprocessed in parallel, and would enable an accurate identification of any infective strains, in acompetitive timescale.The key objectives proposed are:1. Develop & optimise a high throughput molecular tool to be able to effectivelyseparate and identify specific species of Cryptosporidium, present as single and mixedcontaminations;2. Validate the molecular tool using aliquots of real samples and compare the resultsobtained using molecular methods with those obtained by current methods.A molecular tool will enable high throughput routine screening capabilities, allowing a rapidresponse to potential outbreaks. As well as reducing costs and reputation effects following apossible shut-down, this offers savings to the water industry in terms of analytical time.