A primer set and kit for detecting foot-and-mouth disease virus by rt-lamp method and its application

Abstract

The invention relates to a primer set and kit for detecting a foot-and-mouth disease virus through an RT-LAMP method and an application thereof. Six primers of the primer set are positioned at 7776-8050 bp of a whole genome in a highly conserved 3D protein gene of the foot-and-mouth disease virus. The invention provides a kit for detecting the foot-and-mouth disease virus through the RT-LAMP method at the same time. The result can be read according to color reaction during detection application, green or yellowish green reaction shows that a nucleic acid of the foot-and-mouth disease virus ispositive, and orange-yellow or brownish red reaction shows that the nucleic acid is negative; or whether a trapezoidal strip appears or not is analyzed through nucleic acid electrophoresis; and if thetrapezoidal strip appears, a sample is positive, or else, the sample is negative. The primer set and the kit have the characteristics of being high in specificity, high in sensitivity, simple and convenient to operate and low in requirements for instruments and equipment, can relatively well meet the urgent needs for field detection of the foot-and-mouth disease virus at present, and are suitablefor rapid visual detection of the foot-and-mouth disease virus on the scenes, such as grass-roots veterinarians, import and export quarantine departments, farms and slaughterhouses.

Description

technical field [0001] The invention relates to the technical field of biological diagnosis and detection, in particular to a group-specific RT-LAMP detection primer set, kit and method for detecting foot-and-mouth disease virus. [0002] technical background [0003] Foot-and-mouth disease is one of the major animal diseases caused by foot-and-mouth disease virus. It mainly infects cloven-hoofed animals such as pigs, cattle, and sheep. The morbidity and mortality of infected animals are relatively high. The World Organization for Animal Health (OIE) classifies it as a category A animal The first of the epidemic diseases, my country also lists it as a class of animal diseases. Foot-and-mouth disease virus belongs to Picornaviridae, genus of foot-and-mouth disease virus, a total of 7 serotypes (O, A, Asia-1, C, SAT1, SAT2, SAT3 types), more than 70 subtypes, no cross protection between serotypes. The prevention and control of foot-and-mouth disease is a worldwide problem, and ...

AI Technical Summary

Problems solved by technology

In recent years, there have also been researches on the RT-LAMP method for foot-and-mouth disease virus in China, but there are also common problems such as different sensitivities to different serotypes, insufficient detection sensitivity, incomplete coverage, and inability to realize on-site visual detection.

In particular, the existing problems of the current FMD virus RT-LAMP detection method are: 1. The highly conserved fragments of the FMD virus group-specific fragments have not been found. The established general-purpose detection method cannot achieve full coverage of the main epidemic strains, or the sensitivity is low; 2. In the existing research results that realize visualization, it is necessary to open the reaction tube cover to add fluorescent dye after the reaction is completed, which is easy to cause reaction. The product contaminates the environment, and a false positive test result occurs when retesting

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