Developing Readily Deployable Antibody Assays for Canine Distemper Virus in Wild Tigers (Panthera tigris)

Canine distemper disease is emerging as a serious threat to endangered species particularly carnivores including the Amur tiger and leopard. Caused by canine distemper virus (CDV), infected individuals display respiratory and gastrointestinal symptoms, with neurological symptoms developing in the later stages of infection. In recent years, the importance of managing this virus had become a higher priority for conservationists and wildlife veterinarians. Without intervention, endangered populations are at an increased risk of extinction. Current preventative measures involve the vaccination of populations that are in close proximity to vulnerable species, for example domestic dogs in Africa. Yet for accurate monitoring of viral exposure within an area, a simple to use diagnostic device that detects CDV antibodies would be extremely beneficial. Using molecular cloning techniques including restriction enzyme and Golden Gate cloning, a recombinant plasmid was generated that encodes a truncated sequence of the CDV Hemagglutinin glycoprotein. To be ready to use in a diagnostic assay, recombinant protein was produced in bacteria and purification strategies were assessed leading to the utilisation of an inclusion body purification. Appropriate detection methodology was developed utilising Protein A/G via dot blot and preliminary lateral flow assays. Here, investigations into the establishment of a rapid CDV diagnostic tool revealed promising initial data from a basic lateral flow set up, indicating positive detection of the recombinant protein using anti-CDV antibodies and Protein A/G gold conjugates. In summary, this work has set the foundations into future work for lateral flow assays using the CDV Hemagglutinin glycoprotein, alongside the development of a dot blot method. Having such a device would allow for levels of CDV within a population to be known and fitting management approaches put in place to protect species from extinction. The portability and ease of use of the device is important as it would allow rural areas where many endangered species live to have access to such a device.