Method and application for detecting listeria monocytogene by combining isothermal amplification of recombinase polymerase and test strip

Abstract

The invention discloses a method for detecting listeria monocytogene by combining isothermal amplification of recombinase polymerase and a test strip. The method comprises the following steps: selecting an hlyA gene as a detection target gene, designing a primer pair having specificity for the listeria monocytogene, adopting the listeria monocytogene genome DNA as a template, performing the specific isothermal amplification, and obtaining a detection result by utilizing a colloid gold test strip. The method is rapid, simple, high in specificity, high in sensitivity and high in repetition; by adopting the method, the isothermal amplification only needs 10 minutes at 39 DEG C; the detection result of the colloid gold test strip can be made within 5 minutes, the detection result can be observed by eyes, the entire reaction process does not need the expensive experiment instrument and only needs on isothermal instrument, and the method can be applied to the remote regions, the resource-poor regions and the on-site detection. The method is expected to become a simple conventional detection means and has important significance for ensuring the food security.

Description

technical field [0001] The invention belongs to the technical field of microbial detection, and relates to a constant temperature amplification rapid detection technology for Listeria monocytogenes, in particular to a method for detecting Listeria monocytogenes by combining recombinant enzyme polymerase constant temperature amplification (RPA) with a test strip and application. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes) widely exists in nature and is easy to contaminate meat, milk, seafood and vegetables. Because it is easy to form biofilm, it can survive in food for a longer period of time. The World Health Organization pointed out in a report on food poisoning that 4% to 8% of seafood, 5% to 10% of milk and its products, and more than 30% of meat products were contaminated by Listeria monocytogenes. Moreover, since the bacteria can also grow and reproduce in food stored in a refrigerator at 4°C, the potential hazards are further increase...

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[0005] Through the search, no patent publications related to the application of the present invention have been found

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