Quantitative detection method of bovine-derived and porcine-derived components based on droplet digital PCR (polymerase chain reaction) as well as primer, probe and kit

Abstract

The invention relates to the field of molecular biology and discloses a quantitative detection method of bovine-derived and porcine-derived components based on droplet digital PCR (polymerase chain reaction) as well as a primer and probe combination and a corresponding kit used by the method. The sequences of the primer and the probe are SEQ ID NO. 1-6. The method is applied to quantitative detection of bovine components in beef or beef products doped with pork; the defect that real-time fluorescence PCR quantitative detection needs to rely on a standard product is overcome; the method is capable of calculating the accurate content of one certain meat component by accurately measuring nucleic acid copy numbers of two meat components under the condition without any standard product or reference product; the method has extremely important practical significance for cracking down on counterfeit goods in the market and safeguarding the rights of consumers.

Description

technical field [0001] The present invention relates to the field of molecular biology, specifically a method for quantitatively detecting bovine and porcine-derived components using droplet digital PCR technology, as well as the combination of primers and probes used in the method, and the corresponding Reagent test kit. Background technique [0002] In recent years, the donkey-hide gelatin that has been exposed by the media does not contain donkey skin, fake beef, fake pig's trotters, duck meat made of mutton and skewers, American ginseng mixed with dried radish, cottage propolis, fake fish balls, fake bird's nest, The adulteration and sales of fake Cordyceps etc. involved bazaars, restaurants, and even well-known supermarkets and major e-commerce platforms that are deeply trusted by consumers. Faced with these food and health care products that are difficult to distinguish between true and false, consumers are always at a loss. How to identify whether these commodities ...

AI Technical Summary

Problems solved by technology

This technology has the following problems: (1) The difference in the background between the calibrator and the sample will introduce bias and affect the efficiency and measurement response of PCR; (2) The target DNA molecules with low copy number may not be detected by amplification; (3) ) The PCR amplification efficiency of the sample may be different from that of the calibrator; (4) The impurities introduced into the DNA solution during DNA extraction or the degradation of DNA affect the dynamic PCR amplification process

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