Development of an Instrument for Rapidly Detecting Cryptosporidium in Drinking Water

隐孢子虫是一种水生微生物，可引起严重腹泻，对免疫功能低下的个体、婴儿和幼儿可能是致命的。据估计，隐孢子虫污染的饮用水每年在发展中国家导致2.5亿至5亿例，仅在英国就有6万例。隐孢子虫有机体有一层厚厚的外壁，对许多传统的水处理方法具有抵抗力，即使在发达国家，疫情也是一个问题，对人口健康和经济发展产生负面影响——需要对供水进行日常监测。目前的隐孢子虫检测方法昂贵且非常耗时，需要熟练的科学家进行显微镜检查。此外，这些技术缺乏物种和生存能力信息，而这些信息对于做出知情的公共卫生决策至关重要。因此，迫切需要一种能够快速分析饮用水样品中隐孢子虫微生物的存在、种类和生存能力的仪器。在本项目中，我们将开发一种能够快速检测饮用水中隐孢子虫微生物的新型仪器。该仪器将使用拉曼光谱技术，这是一种成熟的激光光谱技术。在拉曼光谱学中，将激光射向感兴趣的样品，一些光子从样品中的分子中非弹性地散射，通过诱导样品分子中的振动而损失能量。散射的光子在波长上发生了位移（正如1930年诺贝尔物理学奖得主钱德拉塞哈拉·文卡塔·拉曼爵士首次观察到的那样），散射光的光谱就像分子的“指纹”，包含了样品分子组成和键结构的高度特定信息。已经开展的工作表明，拉曼光谱可以用来识别饮用水中的隐孢子虫，但目前的仪器对于实际应用来说太慢了。在这个项目中，我们将构建一种新型的拉曼光谱仪，能够同时测量数百个点的拉曼光谱。该仪器将促进实际水测试应用所需的快速测试能力，在几个小时内提供详细的测试信息。

Cryptosporidium is a waterborne microorganism which causes severe diarrhoea and can be fatal for immuno-compromised individuals, infants and young children. It is estimated that Cryptosporidium contamination of drinking water results in 250-500 million cases each year in developing countries and 60,000 in the UK alone. The Cryptosporidium organism has a thick outer wall that is resistant to many conventional water treatment methods, and outbreaks are a problem even in the developed world, negatively impacting population health and economic development - daily monitoring of the water supply is required.Current Cryptosporidium detection methods are expensive and highly time-consuming - requiring microscopic examination by skilled scientists. Furthermore, these techniques lack species and viability information, which is essential to make well-informed public health decisions. There is, therefore, a pressing need for an instrument capable of rapidly analysing drinking water samples for the presence, species and viability of Cryptosporidium microorganisms.In this project we will develop a novel instrument capable of rapidly detecting Cryptosporidium microorganisms in drinking water. The instrument will operate using Raman spectroscopy, a well established laser spectroscopy technique. In Raman spectroscopy, a laser is fired at the sample of interest, some of the photons scatter inelastically from the molecules in the sample, losing energy by inducing vibrations in the sample molecules. The scattered photons are shifted in wavelength (as first observed by Sir Chandrasekhara Venkata Raman, winner of the 1930 Nobel prize for Physics) and the spectrum of scattered light acts as a molecular "fingerprint", containing highly specific information about the molecular composition and bond structure of the sample.Work has already been conducted which demonstrates that Raman spectroscopy can be used to identify Cryptosporidium in drinking water, but current instruments are too slow for real world applications. In this project we will build a new type of Raman spectroscopy instrument capable of measuring the Raman spectra of hundreds of points simultaneously. This instrument will facilitate the rapid testing capability necessary for real world water testing applications, providing detailed test information in a few hours.