Isothermal amplification method for rapid detection of listeria monocytogenes

Abstract

The invention provides a real-time fluorescent quantitative helicase-dependent isothermal nucleic acid amplification method for detecting listeria monocytogenes. Primer pair sequences having specific amplification function on listeria monocytogenes are designed so as to reflect a natural process for simulating the replication of in-vivo DNA at a constant temperature, DNA double strands are solved at a constant temperature by virtue of helicase, the listeria monocytogenes genomic DNA is adopted as a template, specific isothermal nucleic acid amplification is performed so as to achieve the exponential growth of the target sequence, the real-time fluorescent helicase-dependent isothermal nucleic acid amplification detection method is established, detection results are analyzed by virtue of an isothermal amplification curve and melting curve of the genomic DNA so that listeria monocytogenes is rapidly detected. By the method, the disadvantage that the traditional PCR needs to rely on the repeated heating and cooling of an instrument to obtain the single-stranded template is overcome and the method is simple in principle, convenient to operate and listeria monocytogenes in foods can be rapidly, simply, sensitively and specifically detected.

Description

Technical field [0001] The invention belongs to the field of food safety, and in particular relates to a detection method for Listeria monocytogenes in food. Background technique [0002] Listeria monocytogenes, referred to as "Listeria monocytogenes", is a Gram-positive short aseptic bacterium. Listeria has 2 groups and 7 species in total, among which Listeria monocytogenes is an important food-borne pathogen that can cause human diseases. Listeria monocytogenes is widely distributed in nature, and foods such as vegetables, dairy products, seafood, meat, and poultry have all been confirmed as carriers of its transmission. The susceptible populations of this bacteria are mainly pregnant women, newborns, the elderly and Those with immunodeficiency will mainly cause listeriosis, septicemia, meningitis and miscarriage after infection, and the fatality rate is as high as 20% to 50%. In 2000, the World Health Organization listed Listeria monocytogenes as one of the food-borne p...

AI Technical Summary

Problems solved by technology

The newly synthesized double strands are decomposed into single strands under the action of helicase, and used as templates for the next round of synthesis to enter the cyclic amplification reaction, and finally realize the exponential growth of the target sequence, which overcomes the need for repeated instrument upgrades in traditional PCR. Disadvantages of cooling down to obtain single-stranded templates

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