Rapid detection primer group of H7N7 subtype avian influenza virus and use thereof

Abstract

The invention provides a group of primers used for detecting H7N9 subtype avian influenza virus and a method of utilizing the primer group to detect the H7N9 subtype avian influenza virus. The detection method provided by the invention has the advantages of high specificity, high speed, efficient amplification, high sensitivity, convenience for identification, and the like, thereby being capable of being extensively applied in clinic routine detection and epidemiology investigation.

Description

technical field [0001] The invention belongs to the fields of molecular diagnosis and gene diagnosis, and is a microorganism detection reagent, and the detection object of the reagent is H7N9 subtype avian influenza virus RNA. Background technique [0002] H7N9 avian influenza is a new type of avian influenza that was first discovered in Shanghai and Anhui at the end of March 2013. Avian influenza virus belongs to Orthomyxoviridae Influenza A virus genus. Avian influenza A virus particles are pleomorphic, with a spherical diameter of 80-120nm and a capsule. The genome is segmented single-stranded negative-sense RNA. According to the different antigenicities of outer membrane hemagglutinin (H) and neuraminidase (N) proteins, it can be divided into 16 H subtypes (H1-H16) and 9 N subtypes (N1-N9). In addition to infecting poultry, avian influenza A virus can also infect humans, pigs, horses, mink and marine mammals. The subtypes of avian influenza viruses that can infect hu...

AI Technical Summary

Problems solved by technology

However, there are certain disadvantages in the current detection technology: although virus isolation is the gold standard for diagnosis, this method can only be carried out in BSL-3 laboratories, and it is easy to cause the possibility of direct contact infection of operators, which is not conducive to popularization and promotion

In addition, there are related detection methods based on PCR technology. This technology has the disadvantages of complex operation, high detection cost, which invisibly increases the burden on patients, requires high operators, and is not conducive to the development of primary hospitals.

In addition, according to the principle of PCR technology, there are three steps of denaturation, annealing, and extension. The reaction temperature and time of each step are different and have very precise requirements. It even needs 20-40 cycles to complete, so it is necessary Reliance on a relatively expensive PCR temperature cycler for control, which is not conducive to the requirements of on-site diagnostics

Although nucleic acid sequence amplification (NASBA), self-sequence replication (3SR) and strand displacement replication (SDA) are isothermal amplification methods, their amplification specificity for the target sequence is not strong, making it necessary to follow up after amplification. The experimental operation method is used to detect the amplification product, so it has the disadvantage of cumbersome operation

During the concentrated outbreak of pathogens, due to the long period of obtaining monoclonal antibodies, it is not conducive to detection during disease outbreaks

[0005] In order to improve the current disadvantages of H7N9 detection, such as complex operation, reliance on expensive equipment, high detection cost, and high requirements for operators, a method that does not rely on expensive equipment can be developed to detect patient samples within 1 hour, and The H7N9 detection method that can directly observe the results with the naked eye has the characteristics of strong specificity, high sensitivity, and high accuracy rate, making its reaction programmable and can be widely used in clinical testing and epidemiological investigations

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