Taqman-LNA multiplex quantitative PCR method for simultaneously detecting cattle and pig-derived ingredient in meat and meat product, primer probe and kit thereof

Abstract

The invention relates to a Taqman-LNA multiplex quantitative PCR method for simultaneously detecting cattle and pig-derived ingredient in meat and a meat product. The method uses a LNA probe sequence with short sequence and high Tm, under prerequisite that specialty is ensured, stability of a system is effectively increased, a set of multiplex fluorescence quantitative PCR detection system is established, the system can simultaneously detect cattle and pig-derived ingredients, and can detect the cattle-derived ingredient with DNA content being 0.05ng and the pig-derived ingredient with DNA content being 0.05ng in the meat and the meat product, which equals detection of the corresponding animal ingredient with content being about 0.005%. The method has the advantages of high flux, strong specialty, high sensitivity and good application, and can be used for detecting cattle and pig-derived ingredient in meat and the meat product.

Description

technical field [0001] The invention belongs to the technical field of food quality and safety detection, and in particular relates to a Taqman-LNA multiplex fluorescent quantitative PCR method and a primer probe set for simultaneously detecting bovine and pig-derived components in meat or meat products. Background technique [0002] The adulteration of meat or meat products has always been one of the focuses of consumers. Some unscrupulous businesses use low-priced meat products as high-priced meat products for personal gain, or mix low-priced meat products with high-priced meat products. This seriously damages the the interests of consumers. At present, combating the adulteration of meat products has become the focus of the regulatory authorities, and the detection technology of meat or meat products is the basis of supervision. [0003] At present, DNA-based molecular biology methods are the most widely used detection technology for animal-derived components. In partic...

AI Technical Summary

Problems solved by technology

Although the Taqman fluorescent PCR technology overcomes the above-mentioned shortcomings of ordinary PCR, it needs a longer probe sequence to obtain a higher Tm value when designing a probe. Therefore, it is difficult to find the ideal probe sequence, and it is impossible to obtain efficient and specific amplification effects. From time to time, it is found that these methods have certain false positive or false negative results in the detection.

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