Primers, probe composition and kit for rapid identification of nine animal origin ingredients in food or feed, detection method for identification of nine animal origin ingredients in food or feed and application of primers, probe composition, kit and detection method

Abstract

The invention belongs to the technical field of molecular biology and provides primers, a probe composition and a kit for rapid identification of nine animal origin ingredients in food or feed, a detection method for identification of the nine animal origin ingredients in the food or feed and application of the primers, the probe composition, the kit and the detection method. Origin ingredients of the food or feed containing multiple species can be identified rapidly by the primers, the probe composition and the real-time fluorescent PCR (polymerase chain reaction) joint detection kit. The detection method includes: designing a universal primer by taking 16SrDNA as a target gene, and designing specific probes of nine species by designing to construct internal amplification control and designing the specific probes aiming at internal control sequences; performing PCR amplification by three PCR reaction systems; interpreting the origin ingredients of the nine species directly through corresponding fluorescent probe signals and Ct values. The detection method is low in cost, time saving and high in efficiency and can achieve identification of multiple species simultaneously.

Description

technical field [0001] The invention relates to a primer, a probe composition, a kit, a detection method and an application thereof for rapidly identifying nine kinds of animal-derived components in food or feed, and belongs to the technical field of molecular biology. Background technique [0002] The horsemeat scandal in Europe in 2012 caused countries to attach great importance to the authenticity and adulteration of meat. Since 2013, incidents of adulteration of high-grade meat and low-quality meat in China have also occurred frequently. The media have successively reported Shandong Binzhou, Inner Mongolia , Shanghai, Xi'an, Shenzhen and other places investigated and dealt with fake mutton incidents. After investigation, the main problem of meat adulteration in our country is: mixing pork, chicken, duck, horse meat, mouse meat, fox meat, mink meat, etc. into high-end and expensive meat such as mutton and beef, which are easy to breed and more expensive. Cheap meat, shod...

AI Technical Summary

Problems solved by technology

After investigation, the main problem of meat adulteration in our country is: mixing pork, chicken, duck, horse meat, mouse meat, fox meat, mink meat, etc. into high-end and expensive meat such as mutton and beef, which are easy to breed and more expensive. Cheap meat, shoddy, cheating consumers

The current detection of animal-derived components based on multiplex PCR technology mainly relies on the use of agarose gel electrophoresis to distinguish amplified fragments of different lengths after PCR. Patents with application numbers 201510061793.2 and 201510300666.3 both use the melting curve of multiplex RT-PCR Analytical technology to achieve rapid identification of multiple origin components in meat and meat products, but this method cannot distinguish species with close homology of congeners such as foxes and raccoons only through the range of melting curve temperature TM values. The designed primers require strong specificity, and multiple pairs of primers are likely to cause cross-reaction and non-specific amplification

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