Real-time fluorescence RT-HDA (Reverse Transcriptase-Helicase-Dependent Isothermal Amplification) kit and primer for detecting foot-and-mouth disease virus

Abstract

The invention discloses a real-time fluorescence RT-HDA (Reverse Transcriptase-Helicase-Dependent Isothermal Amplification) kit and primer for detecting foot-and-mouth disease virus. A group of primers for detecting the foot-and-mouth disease virus have an oligonucleotides sequence shown by a sequence table SEQ ID No:1 and a sequence table SEQ ID No:2. The kit and the primer have the following advantages that the novel HAD technology is applied to the detection of the foot-and-mouth disease virus, and compared with the existing FMDV (Foot-And-Mouth Disease Virus) detecting method, the HDA detecting method has the advantages of fastness, more convenience, safety and reliability; for the conventional virus separation and serology method, long time is consumed, the bio-safety risk exists; for the HDA method, a natural reproduction mechanism in a body is simulated, the safety is high, the reaction speed is quick, and the detection can be completed by only 2 hours at most; and compared with fluorescence PCR (Polymerase Chain Reaction), the HDA detecting method has the advantages that the program setting is easier, the reaction conditions such as annealing temperature and the like do not need to be explored, and the HDA detecting method is easier to master and operate in the aspect of technical level. Due to the optimization of reaction conditions, according to the real-time fluorescence RT-HDA method, at least 10copies / reaction of viral nucleic acid can be detected. The technology has a great application prospect in the monitoring and diagnosis of the foot-and-mouth disease virus, and the kit and the primer are diagnosis tools with good promotion and application values.

Description

technical field [0001] The invention relates to a real-time fluorescent RT-HDA kit and primers for detecting foot-and-mouth disease virus, belonging to the field of inspection and quarantine. Background technique [0002] Foot-and-mouth disease (FMD) is an acute febrile highly contagious infectious disease caused by Foot-and-mouth disease virus (FMDV). It mainly infects artiodactyls. The mouth, tongue, and Blisters occur on lips, hooves, breasts and other parts, and ulceration forms rotten spots. Humans and non-cloven-hoofed animals can also be infected with the disease, but the symptoms are milder. Because pigs, cattle, sheep and other major domestic animals can be infected with the disease, and can form a large-scale global epidemic, the World Organization for Animal Health (OIE) and the Food and Agriculture Organization of the United Nations (FAO) have listed the disease as a category A zoonotic disease. In my country, foot-and-mouth disease is listed as a class of anim...

AI Technical Summary

Problems solved by technology

At present, international research on real-time fluorescent HDA technology is still in its infancy, and there is no research on real-time fluorescent HDA detection of animal viruses at home and abroad.

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