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, MERS and influenza from symptoms, and even some test kits have serious cross-reactions

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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