Multiple-PCR primer system for quickly testing animal origin ingredients of pigs, sheep and cows and testing method

Abstract

The invention provides a multiple-PCR primer system for quickly testing animal origin ingredients of pigs, sheep and cows and a testing method. The testing method comprises the following steps that the total DNA of a to-be-tested sample is taken as a template, the multiple-PCR primer system composed of a first primer pair, a second primer pair and a third primer pair is utilized to perform a multiple-PCR amplification experiment, and after reacting is completed, a result is judged according to agarose gel electrophoresis; the total DNA template of the sample adopts one or multiple of a pork product, a mutton product and a beef product. According to the multiple-PCR testing method, the authenticity of the pork product, the mutton product and the beef product and whether adulteration exists or not can be quickly and highly specifically tested, the testing efficiency is greatly improved, the time is saved, the testing cost is reduced, and the testing method is particularly suitable for quick anti-counterfeiting authentication of the pork product, the mutton product and the beef product. In addition, the multiple-PCR primer system cooperates with relevant reagents to prepare a kit, use is convenient, and meanwhile possibility of industrial production and application is supplied.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a multiple PCR primer system and a detection method for rapidly detecting animal-derived components of pigs, sheep and cattle. Background technique [0002] People regard food as the first priority, and food safety is the first priority. Eating safe, healthy and green meat products is an important aspect of food quality and safety. However, adulteration and unintentional contamination of animal ingredients often occur in animal-derived raw materials and processed products. A number of meat adulteration incidents recently exposed in the domestic market have aroused public concerns about food safety. Even in Europe, which "has the most stringent food safety system in the world", the phenomenon of "selling horse meat under the guise of a bull's head" is unavoidable, which is worthy of people's deep thinking. With the emergence of meat adulteration forms in an endless stream, the r...

AI Technical Summary

Problems solved by technology

These detection standards are currently mainly focused on the use of traditional conventional PCR technology, which can only perform a single qualitative detection of animal-derived meat and meat products

In the actual testing process of meat food samples, for processed products with unclear or mixed ingredients, it needs to go through multiple testing tests to determine, which has a long cycle, heavy workload and high cost.

In the detection of animal-derived meat and meat products in the future, this type of detection technology has become increasingly incapable of dealing with the special needs of several or even more than a dozen animal-derived products at a time, especially when a large number of processed products enter commercialization In production, more efficient and rapid detection methods are required

[0007] Although multiplex conventional PCR can detect several animal components at the same time, the effect of multiplex PCR in the same reaction tube is often poor, short fragments will be limitedly amplified, and recombination in homologous regions will also produce artificial fragments

Therefore, there are two main defects in the multiplex PCR method: one is that a large number of primers exist in one system at the same time, which is easy to cause non-specific amplification; the other is that due to the difference in amplification efficiency, some target molecules will be preferentially amplified

The limitations imposed by these defects become more pronounced as the number of primer pairs added increases and the number of cycles increases

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