Fluorescently-labeled gene multiplex amplification method for identifying sources of goat, sheep, pig and duck meat simultaneously

Abstract

The invention provides a fluorescently-labeled gene multiplex amplification technology for detecting sources of goat, sheep, pig and duck meat simultaneously. A method comprises the steps that mitochondria 16S rRNA (ribosomal Robonucleic Acid) and Cyt (Cytosine) b genes of a goat, sheep, a pig and a duck serve as target genes, and are subjected to PCR (Polymerase Chain Reaction) multiplex amplification by two pairs of fluorescently-labeled primers; PCR amplification products are detected by a 3730xl full automatic gene analysis meter; and species identification is performed according to 16S rRNA gene amplified fragment length polymorphism. The method fills in a gap of a detection method of the four kinds of meat.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a fluorescent marker gene composite amplification method capable of simultaneously identifying four kinds of meat origins such as goat, sheep, pig and duck. Background technique [0002] Food is food to safety first. Meat safety is a major issue related to people's livelihood. Mutton has less fat, rich protein, high calcium, potassium and VB1, low cholesterol content, high nutritional value and food tonic value, and has been favored by people all the time. In recent years, it has been continuously reported that criminals have mixed a large amount of pork, duck and chicken with a small amount of beef and mutton fat and leftovers to sell as mutton, earning high profits. It has become a "hidden rule" in the circle to make lamb skewers with pork and duck meat with flavor and pigment. Reports about fake mutton continue to appear in the newspapers, which not only brings...

AI Technical Summary

Problems solved by technology

With the improvement of adulteration means, the combination of adulterated species is becoming more and more diverse, among which pork, duck and chicken are the most common in mutton, but there is no simultaneous detection of goat, sheep, pig and duck. The method for growing meat, the present invention uses mitochondrial 16S rRNA and cyt b gene as the target gene, uses two pairs of primers to carry out PCR amplification to the mtDNA16S rRNA of 4 kinds of animals, and the PCR amplification product is detected by a 3730xl automatic gene analyzer. The cyt b gene is used as an internal reference, and the species identification is carried out according to the polymorphism of the amplified fragment length of the 16S rRNA gene, which makes up for the gaps in the above four meat detection methods

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