Development of a rapid and sensitive method to detect the parasite Cryptosporidium in drinking water

  • Lidia D'Alessandro

    Student thesis: Master's Thesis


    Cryptosporidium is a parasite commonly found in water and is the causative agent for the waterborne intestinal disease called cryptosporidiosis.

    Common techniques to detect Cryptosporidium in water include the utilization of filters to trap the oocysts and the examination of the filtered water, observing slides under the microscope. The whole process is time consuming, and the success of the analysis is often linked to good technical and observational skills of the operator.

    In the literature there are several attempts of detecting Cryptosporidium spp. in wastewater using PCR techniques. However, the method appears not to be sensitive enough to give clear results. This research proposes new molecular techniques to detect the presence of Cryptosporidium spp. in drinking water using the innovative method of LAMP to detect and amplify free DNA in reservoir water samples. In contrast to PCR, LAMP utilizes a constant amplification temperature of 65 ˚C and uses 6 primers instead of 2. The utilization of more primers grants a higher specificity compared to PCR and the constant temperature makes the process easier and quicker with the opportunity to test the water sample in the field.

    The results obtained from the analysis of 40 water samples demonstrated how LAMP is clearly able to detect free Cryptosporidium DNA in water samples with a high rate of sensitivity and specificity, greatly reducing the testing time from 2.15 hours needed to perform PCR and electrophoresis, to 30 minutes needed to run LAMP. These results are encouraging on the potential of this method to successfully become the leading method for detecting Cryptosporidium presence in water samples.
    Date of Award2023
    Original languageEnglish
    SponsorsKESSII & Dŵr Cymru Welsh Water
    SupervisorJeroen Nieuwland (Supervisor) & Cerith Jones (Supervisor)

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