Detection method and detection kit of influenza A virus, H1N1 and H3N2 subtype influenza virus

Abstract

The invention discloses a detection method and a detection kit of influenza A virus, H1N1 and H3N2 subtype influenza virus, which has the advantages of specificity, sensitivity, good repeatability, fastness and low cost. A specific primer and a TaqMan probe are designed through sequence alignment for searching a highly conserved area according to a North American variant strain M gene of Influenza A Viruse(H1N1)virus in 2009, an HA gene sequence and the HA gene sequence of Influenza A Viruse(H3N2) epidemic strain, which are published in GeneBank; key reagents such as the probe, the primer and positive control (to structure the influenza A virus, the H1N1 and the H3N2 subtype influenza virus gene recombination clone plasmid) in the detection method are developed; and the influenza A virus, H1N1 and H3N2 subtype influenza virus real-time fluorescence RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method is established through the optimization of various reaction conditions and the tests on the specificity, the sensitivity and the repeatability. The detection method can be used for simultaneously and specifically detecting the influenza A virus, the H1N1 subtype influenza virus and the H3N2 subtype influenza virus in one test.

Description

technical field [0001] The invention relates to a detection method and a detection kit for influenza A virus, H1N1 and H3N2 subtype influenza viruses. Background technique [0002] Influenza (influenza), referred to as influenza, is an acute respiratory infectious disease caused by influenza virus. The disease is highly contagious, spreads rapidly, and its antigens are easy to mutate, especially influenza A virus, whose evolution frequency is higher than that of B and C Influenza virus has caused three worldwide influenza virus pandemics, such as H1N1 (swine type) in 1918, H2N2 type (Asian type) in 1957 and H3N2 type (Hong Kong type) in 1968. At present, the influenza A outbreaks in Mexico and the United States belong to the H1N1 subtype. The H1N1 subtype influenza virus that caused the outbreak is a new variant of influenza virus - the 2009 North American variant, which can infect humans and pigs at the same time. [0003] In nature, the host range of influenza A virus is ...

AI Technical Summary

Problems solved by technology

Classic virus isolation and serological diagnosis methods are time-consuming and laborious, not suitable for clinical testing, do not meet the requirements of biosafety regulations, and cannot meet the requirements of rapid inspection and release at border ports

Ordinary PCR is not only unable to quantitatively detect viruses in disease samples. Electrophoresis is required after the amplification reaction. This is not only not suitable for the screening of large batches of samples, but also easily causes false positives due to operational contamination.

Single-plex fluorescent RT-PCR can only be used for general-purpose detection of influenza, but cannot be used for definitive diagnosis and typing

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