Development of a loop-mediated isothermal amplification technique for sex detection in Cervidae species

Determining sex in Cervidae species is crucial for wildlife management, conservation, breeding programs, disease surveillance, and forensic investigations. While existing genetic sexing methods have proven effective, they often involve time-consuming and resource-intensive processes. In response to the growing need for a more efficient and cost-effective technique, this study aimed to develop a novel loop-mediated isothermal amplification (LAMP) method for sex detection in various deer species. We designed LAMP primers specific to Cervidae based on sex chromosome-located genes, AmelogeninX, and SRY. Analytical specificity was confirmed using 108 samples from red deer, roe deer, and fallow deer, and species specificity was also tested in other deer and mammal species. Sex determination results were fast (40 min) and accurate (100% reliability). This technique has the advantage of allowing researchers to identify the sex of different deer species using the same method. Using the isothermal amplification properties of LAMP, this new method provides a simple, quick, and accurate tool to help researchers, wildlife managers, and forensic investigators with important sex-related questions in cervid populations.