Fluorescent PCR (polymerase chain reaction) detection method, amplifier and probe for raccoon dogs

Abstract

The invention discloses a fluorescent PCR (polymerase chain reaction) detection method, an amplifier and a probe for raccoon dogs. The fluorescent PCR detection method includes supplementing water into 10 microliters of Premix Ex Taq which is a real-time fluorescent PCR system, 0.4 microliter of upstream primers, 0.4 microliter of downstream primers, 0.4 microliter of the probe and 1 microliter of sample DNA (deoxyribonucleic acid) liquid obtained at a step 1 to obtain 20 microliters of liquid; carrying out reaction programs (1) at the temperatures of 95 DEG C for 10 sec, reaction programs (2) at the temperatures of 95 DEG C for 5 sec and reaction programs at the temperatures of 60 DEG C for 23 sec by the aid of a fluorescent quantitative PCR instrument; carrying out 40 cycles. The fluorescent PCR detection method, the amplifier and the probe have the advantages that endogenous components of the raccoon dogs can be accurately and quickly identified by the aid of the primers and the probe which are high in specificity and sensitivity, and the fluorescent PCR detection method, the amplifier and the probe can be effectively used for identifying the authenticity of meat products, fur and the like and can assist relevant departments to crack down severe behavior of meat and meat product fake production and sales.

Description

technical field [0001] The invention relates to a detection method for animal-derived components, in particular to a fluorescent PCR detection method for raccoon dogs and amplification primers and probes thereof. Background technique [0002] In recent years, incidents of adulteration and adulteration of meat and meat products have been frequently exposed by the media, which has become one of the important challenges facing China's food quality control. Criminals sell relatively cheap pork, chicken, and duck as beef and mutton. Recently, it has been reported that unscrupulous traders mix meat such as fox and mink used for fur into beef and mutton for sale. However, these meats are not a regular source of edible meat, and processed and eaten are very risky to health. In addition to foxes and minks, raccoon dogs are another common fur animal, and their meat is also at risk of being sold as other meat. The detection of animal species components is an effective means to combat...

AI Technical Summary

Problems solved by technology

Real-time fluorescent PCR technology can be used to qualitatively detect animal components of raccoon dogs. The inventions of CN103757117A and CN103757116A disclose the preparation and detection methods of rapid detection kits for raccoon dog components in food and feed, but the sensitivity and sensitivity of the primers and probes used in the invention and The specificity is not ideal, resulting in insufficient detection sensitivity and specificity

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