شناسايي گونه هاي گوشتي با استفاده از روش تكثير هم دما به واسطه‌ي حلقه

Meat, as the main ingredient of diets all over the world, is not only a source of nutrition, but the meat industry and its products are a multi-billion-dollar industry. Fraud in meat products has become a worrying issue in the food industry due to its consequences on food safety, economic health and consumer confidence. Meat adulteration is generally defined as the deliberate reversal of meat products for economic gain. In the meat industry, the main method of adulteration involves the substitution of meat of high commercial value with meat of cheaper or less desirable species. Among other things that affect the quality of meat products, meat contamination such as insects and larvae resulting from their eggs also reduce the quality of meat products and affect the health of the consumer. Due to the shortness of the consumption chain, identifying frauds and contaminations is of particular importance. Loop-mediated Isothermal amplification (LAMP) is a very sensitive DNA-based detection method that can detect the target DNA in a short time using a set of 4-6 primers and without the need for a thermocycler. The aim of this research was to design and optimize a diagnostic kit to identify meat species of sheep, edible birds and meat contaminating insects. In order to achieve this goal, the specific primer of the sheep meat species was designed for the cytochrome b gene, the general primer of birds on the GAPDH gene and the general primer of insects for the 28S rRNA gene were designed. DNA of meat tissues and fly larvae were extracted by salt method and participated in the reaction. Reactions for all sets were performed at two levels of specificity eva‎luation and determination of their detection limit, and the results of the reactions were checked in three ways: gel electrophoresis, colorimetry of dsDNA-binding dyes, and real-time PCR. Sheep meat identification primers with a detection limit of 1.7 pg/l distinguished this type of meat from the rest. Chicken, turkey, goose and duck meat species were identified by the general identification primers of bird meat species with a detection limit of 110.8 fg/l. Finally, primers for identification of meat-contaminating insects generally identified the larvae of meat-contaminating flies, which are mainly from the four families of Muscidae, Calliphoridae, Sarcophagidae, and Fanniidae, from edible meat species. The detection limit of this set of primers was measured to be 219 ag/l.

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