Asia holds wilder cat species than any other continent. Asian Big Cats (ABC) parts have become items of illegal trade in Asia. Live along with ABC parts are used for various purposes i.e. pets, entertainment, human consumption, decoration, jewelry, traditional Asian medicine, customary culture, modern fashion and beliefs. All Asian big cats i.e. tiger (Panthera tigris tigris), Indian indigenous Asiatic lion (Panthera leo leo), common leopard (Panthera pardus), snow leopard, (Uncia uncia) and clouded leopard (Neofelis nebulosa) are classified under CITES Appendix I (CITES 2019). All big cats are found in Nepal excluding lion. Nepal is signatory to the CITES and thus prohibits commercial trans-border trade of big cats listed in CITES Appendix. Except common leopard, all other indigenous big cats are highly protected by national law in Nepal (National Parks and Wildlife Conservation ACT 1973). Currently, Bengal tiger is classified as endangered and Asiatic lion, common leopard, snow leopard and clouded leopard as vulnerable under the IUCN Red List. Since these species are given different levels of protection under national and international laws, an effective prosecution requires specieslevel identification but it is a challenging task for the forensic biologists. It demands practical methods that identify species with near certainty when the species identity is in question.