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

隐孢子虫是一种在许多牲畜和野生动物中发现的病原体。它可能通过饮用水和娱乐用水系统传播给人类，并导致严重的长期胃肠道感染，甚至死亡。这种病原体种类繁多，其中只有3到4种已知对人类具有传染性。英国的案例数量相对较多，这是因为许多供水设施的农村性质以及相关的相对较高的降雨量。水处理设施经常使用显微镜监测隐孢子虫卵囊状结构。一旦确定阳性样本，有必要在斯旺西使用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对人类具有感染性。在混合污染物中识别仍然很困难，这是很常见的。这会带来额外的成本和时间，并通过尽可能地关闭处理设施来确保公众不会生病，从而给自来水供应商带来压力。这里提出了一个项目，开发一种能够结合现有方法的两种元素的分子工具。该工具将是高通量的，使许多样本能够被并行处理，并将能够在竞争激烈的时间范围内准确地识别任何感染菌株。建议的主要目标是：1.开发和优化高通量分子工具，以能够有效地分离和识别特定种类的隐孢子虫，以单一和混合污染物的形式存在；2.使用等分真实样本验证分子工具，并将使用分子方法获得的结果与当前方法获得的结果进行比较。分子工具将使高通量常规筛选能力成为可能，从而能够对潜在的爆发做出快速反应。除了在可能的关闭后降低成本和声誉影响之外，这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.