Novel coronavirus, MERS and influenza A/B virus four-in-one rapid detection kit

A technology of influenza B virus and influenza A virus, applied in the biological field, can solve problems such as cross-reaction of detection kits and distinguishing new coronary pneumonia influenza

Pending Publication Date: 2020-10-20
范春雷 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is difficult for ordinary people to distinguish between new coronary pneumonia, M

Method used

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  • Novel coronavirus, MERS and influenza A/B virus four-in-one rapid detection kit
  • Novel coronavirus, MERS and influenza A/B virus four-in-one rapid detection kit
  • Novel coronavirus, MERS and influenza A/B virus four-in-one rapid detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of rabbit anti-new coronavirus S1 protein polyclonal antibody and rabbit anti-MERS virus S1 protein polyantibody

[0030] 1) immunity

[0031] (1) Take a healthy male New Zealand white rabbit with a body weight of about 2.5kg;

[0032] (2) Prepare the purified recombinant new coronavirus S1 protein with PBS (pH=7.0) solution to make 200μg / 850μl, add 150μl 501 immune adjuvant, and immediately after mixing, intramuscularly inject rabbit hind thighs, 500μl on the left and right sides, which is the initial immunization;

[0033] (3) On the 12th day after the initial immunization, use the same method to boost the immunization once;

[0034] (4) Booster immunization once every 10 days with the same method, a total of 3 booster immunizations;

[0035] (5) One week after each immunization, about 100 μl of blood was collected from the ear vein to measure the potency;

[0036] (6) One week after the last immunization, the serum titer should be determined ...

Embodiment 2

[0046] Example 2 Preparation of L-nCov, L-MERS, L-IFVA, L-IFVB and L-RIgG

[0047] Conjugation of ACE2 to carboxylated colored latex. Specific steps are as follows:

[0048] 1) The Rabbitanti-nCov S1 PAb prepared in Example 1 was coupled to the carboxy colored latex according to the EDC / NHS method described in the product manual of the carboxy colored latex to obtain L-nCov;

[0049] 2) Similarly, the Rabbit anti-MERS S1 PAb prepared in Example 1 was coupled to carboxyl colored latex to obtain L-MERS;

[0050] 3) Similarly, anti-IFVA monoclonal antibody (capture type) was coupled to carboxyl colored latex to obtain L-IFVA;

[0051] 4) Similarly, anti-IFVB monoclonal antibody (capture type) was coupled to carboxyl colored latex to obtain L-IFVB;

[0052] Similarly, rabbit IgG was also coupled to carboxyl green latex to obtain L-RIgG, which was used in the test strip quality control system.

Embodiment 3

[0053] Example 3 Preparation of a four-in-one rapid detection kit for novel coronavirus, MERS and influenza A / B

[0054] 1) The L-nCov, L-IFVA and L-RIgG prepared in the above Example 2 were mixed according to the mass ratio of 1:1:1, diluted with PBS (pH=7.0) solution to a total mass concentration of 0.6%, and sprayed on On the release pad of the test strip, dry at 37°C for 12 hours to form the release pad A, and use it for later use; similarly, mix L-MERS, L-IFVB and L-RIgG and spray on the release pad of the test strip, and dry at 37°C for 12 hours , is release pad B, spare.

[0055] 2) Dilute anti-IFVA monoclonal antibody (detection type), recombinant human ACE2 protein and goat anti-rabbit IgG polyclonal antibody (GAR) to 0.5mg / mL with coating diluent (150mM PB, pH 7.4), and apply to membrane Liquid volume of 30 μL / 30 cm was evenly sprayed and drawn on the T1, T2 detection line area and C1 control line area of ​​the nitrocellulose membrane, dried at 37°C for 12 hours, se...

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Abstract

The invention discloses a novel coronavirus, MERS and influenza A/B virus four-in-one rapid detection kit. A release pad of the detection strip A is sprayed with a colored latex labeled anti-novel coronavirus S1 protein polyclonal antibody and a colored latex labeled capture type anti-influenza A virus monoclonal antibody; a release pad of the detection strip B is sprayed with a colored latex labeledanti-MERS virus S1 protein polyclonal antibody and a colored latex labeled capture type anti-influenza B virus monoclonal antibody; and the detection strip A and the detection strip B are assembledon a duplex clamping plate to form the kit. The detection strip A is used for detecting novel coronavirus and influenza A virus, and the detection strip B is used for detecting MERS and influenza B virus. The kit disclosed by the invention is convenient and rapid in detection, can be used for simultaneously judging four viral epidemic diseases which are difficult to distinguish within 3-15 minutes, can be applied to hospital detection, home detection, epidemic disease investigation and large-scale screening diagnosis, and is suitable for global coronavirus and influenza virus epidemic situation monitoring.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a four-in-one rapid detection kit for novel coronavirus, MERS and influenza A / B. Background technique [0002] Coronaviruses belong to the genus Coronavirus of the family Coronaviridae in systematic classification. Viruses of the genus Coronavirus are positive-sense single-stranded RNA viruses with an envelope (envelope), with a diameter of about 80-120nm. Pangolins, dogs, wolves, chickens, cattle, snakes, birds and other vertebrates. The 2019 novel coronavirus (SARS-CoV-2) is currently the seventh known coronavirus that can infect humans, and the remaining six are HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, and SARS-CoV and MERS-CoV. Studies have shown that SARS-CoV-19, SARS-CoV and HCoV-NL63 infect humans through the viral envelope spike glycoprotein (S-protein) mediated interaction between the virus and the ACE2 receptor on the host cell membrane. Common signs after a person i...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/577G01N33/569G01N33/558
CPCG01N33/558G01N33/56983G01N33/577G01N33/583G01N2333/11G01N2333/165G01N2469/10
Inventor 范春雷朱晓进孙刚臧洋李铭夫
Owner 范春雷
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