Multiplex PCR (Polymerase Chain Reaction) primer for simultaneously detecting salmonella, citrobacter, proteus and Edwardsiellas and design method thereof

Abstract

The invention provides a multiplex PCR (Polymerase Chain Reaction) primer for simultaneously detecting salmonella, citrobacter, proteus and Edwardsiellas and a design method thereof, and relates to the detection of salmonellae and the like. A PCR amplified fragment corresponding to a salmonella primer is 526bp, a PCR amplified fragment corresponding to a citrobacter freundii primer is 161bp, a PCR amplified fragment corresponding to a proteus mirabilis primer is 396bp and a PCR amplified fragment corresponding to an edwardsiella tarda primer is 330bp. The design method comprises the following steps of: 1, designing a multiplex PCR primer combination; 2, screening primers in the primer combination and reserving the primers which can not be used for synthesizing primer dimers; 3, judging competitive advantageous and disadvantageous states of the reserved primers and comparing GC% of the primers which are reserved in the step 2, and can not be used for synthesizing the primer dimers with a base number; selecting and judging the primers with the high GC content as the primers with the advantageous competitive states and performing step 5, and otherwise, performing step 4; step 4, screening the primers with the disadvantageous competitive states again; and 5, amplifying the primers with the advantageous competitive states.

Description

technical field [0001] The present invention relates to the detection of Salmonella, Citrobacter freundii, Proteus mirabilis and Edwardsiella tarda, in particular to a multiplex PCR for simultaneous detection of Salmonella, Citrobacter freundii, Proteus mirabilis and Edwardsiella tarda Primers and their design methods. Background technique [0002] Salmonella, Citrobacter freundii, Proteus mirabilis, and Edwardsiella tarda all belong to the Enterobacteriaceae family and can be transmitted through food and wounds. In addition to causing food poisoning, it can also cause multi-system infections in humans and animals. [0003] Salmonella is an early member of the Enterobacteriaceae family. It has two metabolic types: respiratory and fermentative. It exists in warm-blooded and cold-blooded animals, food and the environment. It is a common pathogen that can cause typhoid, typhoid, gastrointestinal related diseases such as inflammation and sepsis. There are many serotypes of Sa...

AI Technical Summary

Problems solved by technology

The improved technology of multiplex PCR simplifies the detection process of food microorganisms, shortens the detection time, reduces the detection cost and improves the sensitivity of food microorganism detection; however, there are some problems in the improved technology of multiplex PCR

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