A screening assay based on PCR amplification of Cytochrome b for species identification of some animals in Cervus

Abstract

Sambar deer (Cervus unicolor) is a protective animal in Thailand. Sambar deer meat has become popular among consumer. Therefore, the smuggling of sambar deer has increased. For the success of prosecution, species identification of meat origin is necessary. An easy-to-perform screening assay for species identification of sambar deer meat is described in this study. The detection of three species of cervids including sambar deer, sika deer (Cervus nippon) and rusa deer (Cervus timorensi) was achieved by using newly designed primers targeting sambar deer cytochrome b (Cyt b) region producing an amplicon of approximately 350 base pairs (bp). The possibility of cross-amplification was prevented by testing with other species of cervids including eld’s deer (Cervus eldi), muntjac (Muntiacus spp.), chital deer (Cervus axis), hog deer (Axis porcinus) and popular consumed meats (pork, beef, chicken, seabass). The limit of detection (LOD) on sambar deer DNA was 31.25 pg. Sensitivity of detection of possible sambar deer in mixed meat species was 2%. Suitability of the screening assay was confirmed on processed meats like frozen meats (-20oC,-80oC) and cooked meats including boiled, steamed, autoclaved, fried, microwave cooked and grilled. Hence, this screening assay can effectively be used for preliminary examination of sambar deer origin in questioned evidence.