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Novel coronavirus antigen and influenza virus antigen combined detection reagent strip and preparation method thereof

A technology for influenza A virus and influenza B virus, which is applied in the field of joint detection reagent strips for novel coronavirus antigens and influenza virus antigens and its preparation, can solve the problems of poor sensitivity of the kits, reduce false positive interference, and achieve high sensitivity , strong specific effect

Pending Publication Date: 2021-01-08
南京佰抗生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects in the prior art that the detection of the new coronavirus and the influenza virus requires two sample additions, the sensitivity of the kit is relatively poor, and there are many problems of missed diagnosis and false positives, and to provide a novel coronavirus The combined detection reagent strip and preparation method for virus antigen, influenza A virus antigen and influenza B virus antigen can provide the qualitative detection results of three viruses in one test at the same time, and has the advantages of fast, simple, high sensitivity and strong specificity

Method used

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  • Novel coronavirus antigen and influenza virus antigen combined detection reagent strip and preparation method thereof
  • Novel coronavirus antigen and influenza virus antigen combined detection reagent strip and preparation method thereof
  • Novel coronavirus antigen and influenza virus antigen combined detection reagent strip and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1 colloidal gold labeling mode

[0030] Such as figure 1 Shown: a new type of coronavirus antigen and influenza virus antigen joint detection reagent strip, characterized in that it includes a bottom plate 2, on which a sample pad 9, a gold standard pad 8, a nitrocellulose membrane 3 and a water-absorbent Paper 1, the surface of the nitrocellulose membrane 3 close to the 8 end of the gold standard pad is sequentially coated with anti-new coronavirus antibody 4, anti-influenza virus antibody 5, anti-influenza virus antibody 6 and quality control C line 7; nitrocellulose The prime surface is coated with rabbit anti-mouse IgG antibody near the absorbent paper end, and colloidal gold-marked anti-new coronavirus antibody, anti-influenza virus antibody and anti-influenza virus antibody are sprayed on the gold pad.

[0031] In conventional antibody colloidal gold labeling methods, antibodies are adsorbed to the surface of colloidal gold through hydrophobic interact...

Embodiment 2

[0042] Example 2: Nitrocellulose membrane coated antibody

[0043]1. Prepare coating buffer: Add 0.15g disodium hydrogen phosphate, 0.04g sodium dihydrogen phosphate, and 5g sucrose in 80ml of ultrapure water, stir well, adjust the pH to 7.5, and dilute to 100ml.

[0044] 2. Coating antibody dilution: Dilute the anti-new coronavirus antibody, anti-influenza virus antibody, anti-influenza virus antibody and rabbit anti-mouse IgG antibody to 1.5mg / ml and 1.0mg / ml with the above coating buffer respectively. ml, 2.0 mg / ml and 1 mg / ml.

[0045] 3. Line coating: wash the film-drawing pipeline of the gold standard biological film spraying instrument HM3035 with cleaning solution and pure water for 20 times respectively; inhale the above four diluted coating antibody solutions into four separate Scribe the pipeline and empty the air in the pipeline; adjust the position of each scribing pen so that the distance between the four lines T1, T2, T3, and C is 4mm, and the T1 line is 0.8mm ...

Embodiment 3

[0047] Example 3: Colloidal Gold Marking and Spray Pad

[0048] 1. Preparation of intermediate products

[0049] Take 50ml of colloidal gold solution in a clean beaker, and use 3% potassium carbonate solution to adjust the pH to 8.5. Add 0.8mg protein A / G under the condition of stirring at 100rpm, keep stirring at room temperature for 30min, centrifuge at 12000rpm for 30min, discard the supernatant and resuspend the pellet in 50ml 20Mm Tris (PH 8.5)

[0050] 2. Antibody labeling

[0051] Take 10ml colloidal gold intermediates in 3 times and place them in 3 clean beakers, add 0.2mg of anti-new coronavirus antibody, anti-influenza virus antibody and anti-influenza virus antibody respectively under the condition of 100rmp stirring, at room temperature After reacting for 30 minutes, centrifuge at 14000rpm for 30 minutes, discard the supernatant and resuspend the above three colloidal gold markers in 1ml resuspension (20mM Tris, 5% sucrose, 1% BSA pH 8.5)

[0052] 3. After mixin...

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Abstract

The invention discloses a novel coronavirus antigen and influenza virus antigen combined detection reagent strip which comprises a bottom plate. A sample pad, a gold-labeled pad, a nitrocellulose membrane and absorbent paper are sequentially pasted on the bottom plate, and the surface of the nitrocellulose membrane is sequentially coated with an anti-novel coronavirus antibody, an anti-influenza Avirus antibody and an anti-influenza B virus antibody. A rabbit anti-mouse IgG antibody coats the end close to the absorbent paper. A colloidal gold labeled anti-novel coronavirus antibody, an anti-influenza A virus antibody and an anti-influenza B virus antibody are sprayed on the gold-labeled pad. The binding protein A / G is marked on the surface of the colloidal gold, the protein A / G is specifically bound with the Fc end of the antibody, so that the ideal conformation of Fab end abduction is constructed, and meanwhile, the binding of the protein A / G with the Fc can reduce the false positiveinterference of rheumatoid factors on a detection structure. The reagent strip can simultaneously realize qualitative detection of the novel coronavirus antigen and the influenza antigen in one test,and is convenient to use, good in sensitivity, high in specificity and short in detection time.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a colloidal gold kit for the joint detection of novel coronavirus antigens, influenza A virus antigens and influenza B virus antigens and a preparation method thereof. Chromatography and colloidal gold surface antibody-directed labeling technology realize the combined qualitative detection of novel coronavirus antigens, influenza A virus antigens and influenza B virus antigens in samples. Background technique [0002] Influenza (flu) and COVID-19 are both contagious respiratory diseases, but they are caused by different viruses. COVID-19 is caused by infection with a novel coronavirus (2019-nCoV, also known as SARS-CoV-2), while influenza is caused by infection with the influenza virus. Because some of the symptoms of influenza and COVID-19 are similar, it may be difficult to tell the difference from symptoms alone, so testing may be needed to confirm the diagnosis. [0003] At present...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/58G01N33/53
CPCG01N33/56983G01N33/558G01N33/585G01N33/5306G01N2333/165G01N2333/11G01N2469/10
Inventor 芮兵
Owner 南京佰抗生物科技有限公司
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