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

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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.