Method for rapidly detecting shigella dysenteriae

Abstract

The invention belongs to a rapid detection technology of food-borne pathogens, in particular to a loop-mediated isothermal amplification (LAMP) method for detecting shigella dysenteriae DNA. The method comprises the preparation of reagent comprising sample processing reagent, LAMP reactive reagent, a large fragment of Bst DNA polyase, agarose, bromophenol blue loading buffer solution and ethidium bromide solution and the procedures of the detection of amplification products and sediment generation. The method comprises the following steps: firstly, designing a specific primer; secondly, extracting sample DNA; and thirdly, preparing a reaction system. Compared with the currently adopted traditional bacterial culture method, an immunological detection method and other molecular biology methods, the method has the advantages of simple and rapid operation, no special instrument and reagent, sensitivity and specificity and can be applied to the rapid detection of shigella dysenteriae in foods and other samples.

Description

technical field [0001] The invention relates to the use of a loop-mediated isothermal DNA amplification technique (loop-mediated isothermal amplification, LAMP) in detecting Shigella. Background technique [0002] Shigella spp. is a class of Gram-negative bacilli, and is the most common pathogen of human bacillary dysentery, commonly known as Shigella. Shigellosis is often food-borne or water-borne. Shigella can spread rapidly in crowded and unsanitary conditions, and is often found in places where a large number of people gather, such as restaurants and canteens. The most common reasons for the prevalence of food-borne Shigella are bacillary dysentery in food processing workers or contamination of food by carriers, poor personal hygiene of people in contact with food, and storage of contaminated food at inappropriate temperatures. [0003] At present, there are mainly bacteriological examination and immunological methods to identify Shigella. The former is a national stan...

AI Technical Summary

Problems solved by technology

These methods can amplify the target nucleic acid to a similar order of magnitude and have higher detection limits, 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 that must use expensive modified nucleotides as substrates for the amplification reaction