RPA-IAC primer for detecting vibrio mimicus and method

Abstract

The invention provides a primer pair and an internal amplification control sequence. The primer pair comprises a nucleotide sequence as shown in SEQ ID NO:1 and a nucleotide sequence as shown in SEQ ID NO:2, and the internal amplification control sequence comprises a nucleotide sequence as shown in SEQ ID NO:3. The invention further provides the application of the primer pair and the internal amplification control sequence to the preparation of products for the detection or aided detection of vibrio mimicus, and a method for detecting the vibrio mimicus, based on a RPA-IAC technology. Experiments prove that the RPA-IAC primer is good in specificity, high in sensitivity and short in detection time, does not need special instruments, and is wide in application scope, and the RPA-IAC detection method for vibrio mimicus, based on the primer matched with an internal amplification control, is sensitive, accurate, convenient and rapid, and has a guiding significance on quarantining imported and exported cargoes and products.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a method for detecting vibrio mimicus, a primer and a kit. Background technique [0002] Mimic Vibrio is a Gram-negative Vibrio with a single flagella and power. Like Vibrio parahaemolyticus and Vibrio cholerae, it belongs to the normal marine flora and widely exists in river water, seawater and seafood in nature. The strain grows well in nutrient broth containing no sodium or only 1% sodium chloride and can be isolated from aquatic animals. [0003] In the United States, North America, New Zealand, Bangladesh, and African countries, Vibrio mimicus has been reported to cause disease. It is also reported in Fujian, Beijing, Shanghai, Jiangsu, and Zhejiang in China. It is mostly sporadic or outbreaks. Many; the incubation period is 3 to 72 hours, with an average of 24 hours; the main symptoms are diarrhea, vomiting, abdominal pain, and watery stool or mucus and bloody stool; th...

AI Technical Summary

Problems solved by technology

Among them, the VIDAS-CHL method, EIA method, colloidal gold test paper method and API method all have defects such as poor specificity and low sensitivity, and have not been widely used; LAMP method, as a new detection method, has greatly improved the detection method. Efficiency, which reduces the cost of detection, but produces a higher false positive rate

As a highly sensitive, fast and simple detection method, PCR technology has been widely used in many fields, especially in the detection of pathogens, it has achieved revolutionary results and has become a gold standard for rapid detection of nucleic acids. On this basis, it has developed DNA probe technology, real-time fluorescent PCR technology and PCR combined with denaturing high-performance liquid chromatography (DHPLC) technology, etc., and the design of PCR primers has become a key factor restricting the success of this type of detection method. Conventional PCR primer design not only requires Repeatedly compare the specificity of the primers, and it is necessary to optimize the parameters and reaction conditions of the primers, especially the annealing temperature is repeatedly optimized to prevent non-specific amplification. Under the constraints of these conditions, the selected primers can easily affect the amplification. Increased validity and specificity

In addition, the price of fluorescence detection equipment is generally high, which also limits the popularization and application of this type of detection method to a certain extent.

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