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

Abstract

The invention discloses a real-time fluorescence RT-HDA (Reverse Transcriptase-Helicase-Dependent Isothermal Amplification) kit and a primer for detecting avian influenza virus. A group of primers for detecting the avian influenza 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 HDA technology is applied to the detection of the avian influenza virus, and compared with the existing AIV (Avian Influenza 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 2h 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 avian influenza 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 avian influenza virus, belonging to the field of inspection and quarantine. Background technique [0002] Avian Influenza (AI) is an infection and / or disease syndrome in poultry caused by Orthomyxoviridae, Influenzavirus, and Influenza A viruses. Avian influenza has a long history. It was first discovered in 1878. Perroncito first reported the occurrence of avian influenza in Italy. It has been listed as a severe infectious disease by the International Veterinary Office (OIE). As one of the most important diseases with potential threats, my country has also listed highly pathogenic avian influenza (HPAI) as a first-class animal infectious disease for prevention and control. In recent years, there have been avian influenza epidemics in many areas of my country, which have brought different degrees of blows to the poultry industry. So far, there have been many fatal inc...

AI Technical Summary

Problems solved by technology

However, there are still some shortcomings in the traditional PCR technology: expensive equipment is needed, especially the fluorescent PCR instrument is expensive; it is easily affected by many factors; the amplification time often takes several hours

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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