Method for fast detecting Saimonella

Abstract

The invention belongs to the quick detection technology of food-borne pathogenic bacteria, and in particular relates to a detection method of loop-mediated isothermal amplification (LAMP) for salmonella. The detection method comprises the preparation of a reagent comprising a sample processing reagent, an LAMP reaction reagent, a large segment of BstDNA polymerase, agarose, bromophenol blue sampling buffer solution and ethidium bromide solution and the detection procedures of the detection an amplified product and the generation of deposition. The detection method comprises the following steps: firstly designing a specific primer; secondly extracting DNA from a sample; thirdly, establishing a reaction system. Compared with the traditional bacterial culture method, the immunity detection method and other molecular biology method adopted presently, the method has the advantages of simple operation, quickness, no special instrument reagent and specific sensitivity. The invention can be applied to the quick detection of the salmonella in foods and other samples.

Description

technical field [0001] The invention relates to the application of species-specific primers in the detection of Salmonella by loop-mediated isothermal DNA amplification reaction. These primers can be used to detect Salmonella contamination in food. Background technique [0002] Salmonella (Salmonella spp.) is a kind of Gram-negative intestinal pathogenic bacteria that is widely distributed in nature and can cause human and animals to become ill. It can cause human gastroenteritis, typhoid fever, sepsis and other symptoms . At present, more than 2,000 serotypes have been isolated in the world, and 216 serotypes have been found in my country. According to the statistics of the National Foodborne Disease Surveillance Network, microorganisms contaminated in food are one of the main factors causing foodborne diseases. In the past ten years, Salmonella has always been the most common and main pathogenic factor in foodborne diseases caused by microorganisms in my country, especi...

AI Technical Summary

Problems solved by technology

These methods can amplify the target nucleic acid to a similar order of magnitude, have higher detection limits and are all about one hour, but they still have some defects that cannot be overcome

Some of them require sophisticated instruments to amplify, and some require sophisticated methods to detect the amplified products because of the low specificity of the selection of the target sequence

Despite its simplicity, the need for high-precision thermal cyclers keeps PCR from being a powerful technique that is widely used

On the other hand, NASBA and 3SR that do not use a thermal cycler have greatly reduced specificity, mainly because a relatively low temperature of 40°C must be used for amplification

SDA largely overcomes these shortcomings by using four primers to amplify under isothermal conditions, but there is still a thin link: expensive modified nucleotides must be used as substrates for the amplification reaction