Double-antibody sandwich method for quickly detecting novel H1N1 influenza virus

Abstract

The invention discloses a double-antibody sandwich method for quickly detecting a novel H1N1 influenza virus, comprising the following specific steps: designing the HA1N end spherical sub-gene primer of an amplified-expression novel H1N1 influenza virus; carrying out PCR (Polymerase Chain Reaction) amplification; connecting a target segment with a carrier pET100 / D-TOPO (trioctyl-phosphine oxide), and building a recombinant plasmid; converting an expression cell (escherichia coli BL21); expressing IPTG (isopropyl beta-D-thiogalactopyranoside) induced recombinant protein; analyzing the antigenicity of a purified product by indirect ELISA (enzyme-linked immuno sorbent assay) and Western blottin; purifying the recombinant protein by a HisTrapTMHP (histidine) affinity adsorption column purification system; preparing rabbit serum by a recombinant protein immune rabbit; carrying out ELISA coating by the prepared rabbit serum; taking the rabbit resisting H1N1 influenza virus hemagglutinin protein marked by a horse radish peroxidise as a second antibody; and building a double-antibody sandwich method for detecting the novel H1N1 influenza virus antigen. The method can be used for carrying out quick and effective antigen diagnosis, is convenient to use and accurate to detect, and is suitable for being popularized and applied to areas, such as border ports and the like.

Description

technical field [0001] The invention relates to a double-antibody sandwich method for rapid detection of novel H1N1 influenza virus. Background technique [0002] Influenza A (H1N1 Influenza A (H1N1) virus) is a highly contagious respiratory virus that has been listed as a weapon of bioterrorism. The virus of the 2009 influenza pandemic was a new type of H1N1 virus. This influenza caused a worldwide pandemic and caused great harm to human health. It is imminent to establish a rapid detection method to prevent the occurrence of influenza disease. At present, the methods for detecting the novel H1N1 influenza virus mainly include virus isolation and culture, reverse transcription polymerase chain reaction (RT-PCR), serological diagnosis and antigen detection. However, the isolation and identification of influenza virus takes a long time and costs high, and needs to be carried out in biosafety laboratories or national designated research institutions, while RT-PCR relies on sp...

AI Technical Summary

Problems solved by technology

However, the isolation and identification of influenza virus takes a long time and costs high, and needs to be carried out in biosafety laboratories or national designated research institutions, while RT-PCR relies on specific equipment, good laboratory conditions and high-quality techniques Therefore, it is urgent to study and establish a rapid and effective antigen diagnostic technology for new H1N1 influenza virus

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