Weissella viridescens rapid detection method based on droplet digital PCR

Abstract

The invention discloses a Weissella viridescens rapid detection method based on droplet digital PCR. The method comprises the following steps: firstly, extracting genome DNA of to-be-detected sample enrichment broth, and performing diluting for later use; designing a specific primer and a probe by utilizing a conservative DNA sequence of a Weissella viridescens genome; then carrying out droplet digital PCR amplification and detection on the genome DNA of the to-be-detected sample enrichment broth by using the primer and the probe; after the PCR amplification, detecting each droplet one by oneby using a droplet analyzer, wherein the droplet with a fluorescence signal is interpreted as 1, the droplet without the fluorescence signal is interpreted as 0; and finally, calculating the concentration or copy number of to-be-detected target molecules according to the Poisson distribution principle and the proportion of positive droplets. The number of DNA molecules can be directly obtained through droplet digital PCR, and absolute quantification can be conducted on Weissella viridescens in a sample. The method has the advantages of high sensitivity, accurate quantification, wide detectionlinear range, good specificity and moderate cost.

Description

technical field [0001] The present invention relates to a kind of Weissella viridans ( Weissella viridescens ) droplet digital PCR rapid detection method. Background technique [0002] During the process of segmentation, processing, transportation and retailing, meat is easily polluted by microorganisms in the surrounding environment due to its rich nutrition and high water activity. Meat spoilage is the result of microbial proliferation and metabolism. The proliferation of microorganisms is the most direct cause of meat spoilage. In addition to a series of changes in nutrients such as protein and fat, the spoilage of meat changes its color from bright red and dark red to dark brown or even dark green, and loses its luster. Sticky, produce putrid smell, even mildew, spoiled meat completely loses its processing and edible value. Weissella viride is one of the dominant bacteria that restricts the shelf life of low-temperature meat products, and it is meaningful to develop a ...

AI Technical Summary

Problems solved by technology

However, so far, there have been no reports in China that are conducive to the rapid detection of Weissella viridans by droplet digital PCR method