A method for specific detection of H1 influenza A subtype by combining RPA with immunofluorescence chromatography
A technology of influenza A and fluorescein, which is applied in the field of specific detection of H1 influenza A subtype by combining RPA with immunofluorescence chromatography, can solve problems such as inability to conduct detection anytime and anywhere, health hazards of the crowd, and long testing time
Active Publication Date: 2022-05-03
SICHUAN MACCURA BIOTECH CO LTD
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Problems solved by technology
The epidemic of influenza A H1N1 in 2009 caused great harm to the global economy and the health of the population. Experience in the prevention and control of influenza A indicates that the existing traditional rapid detection technology can no longer meet the needs of quarantine inspection at border ports. Therefore, it is urgent to develop rapid detection methods for influenza A
However, to use this method for testing, samples need to be sent to regional or central laboratories for testing, and testing cannot be done anytime and anywhere, mainly because this testing method requires professional testing personnel, equipment and complicated operating procedures, and the testing time Long (50-90min), so not suitable for point-of-care testing (POCT)
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Embodiment 1
[0043] 1. Materials and methods
[0044] 1.1 Materials
[0045] Template sequence: Search lectin genes in NCBI database: H1 (S67220.1), H5 (AY854190.1) and H7 (AJ493472.1); neuraminidase genes: N1 (AB539741.1) and N9 (CY179917. 1), send these five gene sequences to Sangon Bioengineering (Shanghai) Co., Ltd. for synthesis.
[0046] Common primer design: Search 17 variants of lectin gene (H1-H17) and 10 variants of neuraminidase gene (N1-N10) from the NCBI database, search 6 items for each variant, perform sequence alignment, and find out which of the variants The variable regions between the variants are then compared to find the conserved regions within the variants, and then the intersection of the two regions is used as the template sequence to design specific primers, and 5 pairs of primers are designed for each gene, and the designed primers are sent to Sangon Bioengineering (Shanghai) Co., Ltd. was used for the synthesis.
[0047] Table 1 Primers for Amplification of I...
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The invention belongs to the field of molecular biology detection, and specifically discloses a method for specifically detecting H1 influenza A subtype by combining RPA with immunofluorescence chromatography. The present invention takes the H1 gene detection of influenza A virus as an entry point to establish a set of immunofluorescence chromatography technology based on constant temperature amplification technology, which can carry out isothermal amplification at normal temperature, and only needs the specific primer (SEQ ID NO.1‑2) are modified separately to meet the requirement of one reagent for detecting H1 influenza A subtype. The constructed influenza A detection technology platform can be applied to the rapid screening of influenza A at ports to control the incoming and outgoing of influenza A, which has important application and promotion value.
Description
technical field [0001] The invention belongs to the field of molecular biology detection, and in particular relates to a method for specifically detecting H1 influenza A subtype by combining RPA with immunofluorescence chromatography. Background technique [0002] With the continuous deepening of economic globalization and the continuous development of international trade, the international flow of vehicles, goods, and personnel has become increasingly frequent, and border ports have become an important way to spread infectious diseases and their vectors. In 2009, the epidemic of H1N1 influenza A caused great harm to the global economy and the health of the population. The experience of influenza A prevention and control suggests that the existing traditional rapid detection technology can no longer meet the needs of quarantine inspection at border ports. Therefore, it is urgent to develop rapid detection methods for influenza A. [0003] Hemagglutination inhibition test (Hm...
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IPC IPC(8): C12Q1/70C12Q1/6804C12N15/11
CPCC12Q1/6804C12Q1/701C12Q2531/119C12Q2563/107C12Q2563/149
Inventor 刘东泽赵书阁姜勇
Owner SICHUAN MACCURA BIOTECH CO LTD