Identifying Snow Leopards, Tigers and other Endangered Species by developing a low-cost DNA field test kit

Methods

Summary

Using the snow leopard and tiger as a model, we aim to create a field kit that will easily and effectively identify species from fecal and other samples rapidly and without the need for expensive sequencing equipment. The method will be developed using feces collected from captive snow leopards and tigers and will also include fecal material from other carnivore species commonly confused with these species, for comparison. We will also adjust the method to identify bone and skin samples from museum specimens. We will develop and test two DNA-based approaches: 1) Litmus paper DNA detection and 2) Recombinase Polymerase Amplification (RPA). Both methods will remove the need for on-going analyses in laboratories (e.g. Polymerase Chain Reaction (PCR)) for surveillance purposes, producing a colorimetric response when species-specific DNA is detected.

The rigour of each method for accurate species identification will also be tested on samples of different age and DNA quality to mimic degradation in the field to establish temporal limits for genetic surveys.

Once developed and perfected in the laboratory, the second phase of this study will involve taking the kit into the field where it can be validated and its effectiveness on the ground tested against traditional PCR methods. The Centre for Molecular Dynamics, Kathmandu (CMDN) who are involved in the study of snow leopards and tigers in Nepal will be partners in the project and will be closely involved with the collection of samples in the field and with amplification of species-specific sequences in the laboratory. Capacity building of the CMDN team in Nepal is an integral part of the project.