Rapid diagnosis kit of enterocolitis yersinia genus gene based on loop-mediated isothermal amplification technique

Abstract

The invention discloses a kit for quickly diagnosing Yersinia genes of enterocolitis and a detection method thereof. The kit consists of two pairs of primers, DNA polymerase, a stabilizing solution, a reaction liquid, a sample pre-treatment fluid, a colored solution and a positive control solution, wherein the seven solutions are placed in a container respectively. The kit for quickly diagnosing the genes applies six segments and four primers, and can judge whether a drone substance exists or not according to the fact whether amplification is performed, so as to have high specificity. The kit for quickly diagnosing the genes is quick and high-efficiency, has high sensitivity, can perform amplification reaction only with a constant temperature, and does not require a special reagent and special equipment, so as to have low detection cost. Moreover, the kit for quickly diagnosing the genes is simple and convenient in identification; pyrophosphate ions separated out from dNTP are combined with Mg<2+> in a reaction solution to produce by-products - magnesium pyrophosphate deposits which can be observed and identified by naked eyes; and the color development difference is more obvious and reliable due to negative and positive results after addition of the colored solution.

Description

technical field [0001] The invention relates to a biological detection reagent, in particular to a rapid diagnosis kit and a detection method for the Yersinia enterocolitica gene. Background technique [0002] At present, there are many detection methods for Y. enterocolitica, ranging from the national standard (GB / T4789.8-2003) based on the isolation and identification of pathogenic microorganisms, morphological identification and automatic biochemical identification, to the immunology of specific proteins. Detection technology, nucleic acid probe, polymerase chain reaction (PCR) technology and other molecular biology detection methods [Food Safety Testing and Modern Biotechnology, Chemical Industry Press, 2004]. Among them, the detection of pathogenic nucleic acids has greatly improved in terms of rapidity, safety, accuracy and sensitivity. These new technologies try to break through the traditional microbiological detection modes such as morphology and biochemical reactio...

AI Technical Summary

Problems solved by technology

Among them, isothermal amplification (Isothermal Amplification) nucleic acid rapid detection technology is a great progress in pathogenic nucleic acid detection technology. The established loop-mediated isothermal amplification technology (loop-mediated isothermal amplification of DNA, LAMP) has many advantages. However, there is no gene rapid diagnostic kit for detection of Y. enterocolitica using loop-mediated isothermal amplification technology.