Identification of sheep ( Ovis aries ) meat by alkaline lysis-loop mediated isothermal amplification technique targeting
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ORIGINAL ARTICLE
Identification of sheep (Ovis aries) meat by alkaline lysis-loop mediated isothermal amplification technique targeting mitochondrial D-loop region T. Mounika1 • P. S. Girish2 • M. Shashi Kumar1 • Aparana Kumari2 Sujatha Singh1 • Nagappa S. Karabasanavar3
•
Revised: 30 September 2020 / Accepted: 8 October 2020 Ó Association of Food Scientists & Technologists (India) 2020
Abstract Identification of meat species origin using reliable techniques is a critical requirement for ensuring label compliance, protection of consumer preference and prevention of fraudulence in the meat trade. Although a plethora of protein and DNA based meat species identification techniques are in vogue, need for rapid test suitable for under-resourced laboratories catering point-of-care (PoC) services was construed. Present study deals with development of rapid sheep (Ovis aries) meat identification technique using DNA extraction by alkaline lysis (AL) and loop-mediated isothermal amplification (LAMP) technique. The AL–LAMP specifically amplifies sheep-specific signal of mitochondrial D loop region under an isothermal temperature of 60 °C with an analytical sensitivity of 0.5 ng sheep DNA. The test was highly specific to sheep and performed well even in the presence of DNA of closely related meat animal species such as goat, cattle, buffalo and chicken. The novel primers designed for the AL– LAMP successfully detected sheep meat in raw and cooked meat samples heated up to 121 °C for 30 min. Sheepspecific AL–LAMP assay could detect 0.1% mutton-inbeef adulteration. Novel AL–LAMP assay being simple, rapid and reliable for sheep meat authentication in just
& P. S. Girish [email protected] 1
College of Veterinary Science, Rajendranagar, Hyderabad, Telangana State 500030, India
2
ICAR – National Research Centre on Meat, Chengicherla, Hyderabad, Telangana State 500092, India
3
Karnataka Veterinary, Animal and Fisheries Sciences University, Veterinary College, Gokula Campus, Vidyanagar, Hassan, Karnataka 573202, India
120 min; hence, it could be conveniently used by terminal laboratories engaged in rendering on-site or PoC services. Keywords Sheep meat Alkaline lysis D loop Isothermal amplification Speciation
Introduction Meat is regarded as one of the nutritious food commodities as it is a rich source of proteins, vitamins and minerals (Vandendriessche 2008). Demand for meat and meat products has shown increasing trend consequent to the growing population, raising disposable income and urbanization. Consumers are now concerned about the quality and safety of the meat they eat and demand prompt labeling of origin of species of meat they buy from the market (Delgado 2003). Nevertheless, mounting meat prices prompt offenders to indulge in misrepresentation of costly meats with cheaper alternatives (Shears 2010); surprisingly, a recent study indicated adulteration of mutton with dog meat (Vaithiyanathan et al. 2018). Fraudulent practice of misrepresentation and mislabeling of costly meats jeopardize consumer sentime
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