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Colloidal gold immunochromatography kit for rapidly detecting novel coronavirus IgM antibody and preparation method thereof

A coronavirus and immunochromatography technology, applied in the field of biomedicine, can solve problems such as insufficient detection efficiency of suspected cases, and achieve the effects of low cost, easy product use, and simple preparation process.

Pending Publication Date: 2020-06-12
中山生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] So far, there are still many suspected cases that have not been diagnosed due to insufficient detection efficiency and other reasons. How to quickly screen suspected cases on a large scale so that clinicians can quickly judge whether it is a new type of coronavirus pneumonia, so as to facilitate rapid isolation and prevent infection to more people , is an urgent problem to be solved

Method used

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  • Colloidal gold immunochromatography kit for rapidly detecting novel coronavirus IgM antibody and preparation method thereof
  • Colloidal gold immunochromatography kit for rapidly detecting novel coronavirus IgM antibody and preparation method thereof

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preparation example Construction

[0038] 1. Screening of preparation conditions for gold-labeled N protein

[0039] 1.1. Determination of the particle size of colloidal gold

[0040] Preparation principle: Colloidal gold is produced by trisodium citrate reduction method. The size of gold particles is greatly affected by factors such as the concentration of trisodium citrate. By changing the concentration of trisodium citrate, colloidal gold with different particle sizes can be produced.

[0041] Test method: Take 2mL of 1% chloroauric acid (HAuCl4) solution, add it to 100mL of water, heat to boil, then add 1-8mL of 1% trisodium citrate, mix and boil for 5 minutes until the color does not change.

[0042] Experiment on the labeling effect of colloidal gold particles with different particle sizes: Colloidal gold particles with different diameters were prepared by the above method, and the new coronavirus nucleocapsid protein containing 6×His tag was labeled to make reagents, and the quality control was performed w...

Embodiment 1

[0075] Embodiment 1, the preparation method of the colloidal gold immunochromatography kit of rapid detection novel coronavirus IgM antibody:

[0076] S1. Production of gold standard pad 3: Take 2mL of 1% chloroauric acid and add it to 100mL of aqueous solution, heat to boiling, then add 2mL of 1% trisodium citrate and mix well, boil until the color of the colloidal gold solution changes from blue to purple to purple, then cool To obtain colloidal gold; take 1mL of colloidal gold and add 100μL of 0.1M, pH7.2 PB, mix well, add 0.1M NaOH solution to adjust the pH value to 7.2, add 15μg of the new coronavirus nucleocapsid protein containing 6×His tag and mix well , then add 100 μL of 10% bovine serum albumin and mix well to obtain the gold standard solution; centrifuge the gold standard solution, discard the supernatant, add gold standard complex solution (0.01M PB+1%BSA) containing 20% ​​sucrose to dissolve, To obtain the gold-labeled N protein; spray the gold-labeled N protein ...

Embodiment 2

[0079] Embodiment 2, the preparation method of the colloidal gold immunochromatography kit of rapid detection novel coronavirus IgM antibody:

[0080]S1. Production of gold label pad 3: Take 2mL of 1% chloroauric acid and add it to 100mL of aqueous solution, heat to boiling, then add 3.5mL of 1% trisodium citrate and mix well, boil until the color of the colloidal gold solution changes from blue to purple to purple, Cool to obtain colloidal gold; take 1mL of colloidal gold and add 100μL of 0.1M, pH7.2 PB, mix well, add 0.1M NaOH solution to adjust the pH value to 7.6, add 20μg of the new coronavirus nucleocapsid protein containing 6×His tag and mix well Then add 100 μL of 10% bovine serum albumin and mix to obtain the gold standard solution; centrifuge the gold standard solution, discard the supernatant, and add gold standard complex solution (0.01M PB+1%BSA) containing 20% ​​sucrose to dissolve , to obtain the gold-labeled N protein; spray the gold-labeled N protein on the go...

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Abstract

The invention discloses a colloidal gold immunochromatography kit for rapidly detecting a novel coronavirus IgM antibody and a preparation method thereof, and relates to the field of biomedicine. Whether an anti-novel coronavirus nucleocapsid protein IgM antibody exists in human serum or blood plasma or not is qualitatively detected by adopting an antigen-antibody sandwich method and a colloidal gold immunochromatography principle; colloidal gold is used for labeling the novel coronavirus nucleocapsid protein containing the 6*His label to be combined to form gold-labeled N protein, and then the gold-labeled N protein is adsorbed on the gold-labeled pad; novel coronavirus nucleocapsid protein is used as an indication marker, the detection line of the NC membrane is coated with a mouse anti-human u chain monoclonal antibody, and the quality control line of the NC membrane is coated with a mouse anti-6*His monoclonal antibody; and qualitative detection of the anti-novel coronavirus nucleocapsid protein IgM antibody in blood is realized by adopting a sandwich method. The kit can conveniently detect whether an anti-novel coronavirus nucleocapsid protein IgM antibody exists in blood or not, and is high in sensitivity, short in detection time and high in interference resistance, so that a clinician can quickly judge whether the novel coronavirus is diagnosed or not.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a colloidal gold immunochromatography kit for rapid detection of novel coronavirus IgM antibodies and a preparation method thereof, specifically a colloidal gold immunochromatography method applying the principle of a sandwich method to realize anti-inflammatory effects in blood. A kit for the qualitative detection of the IgM antibody to the new coronavirus nucleocapsid protein. Background technique [0002] The common signs of the new coronavirus (2019-nCoV) are fever, fatigue, dry cough, and gradual dyspnea. Some patients have mild symptoms, and there are even asymptomatic patients without clinical symptoms. It has the characteristics of human-to-human transmission, the general incubation period is 1-14 days, and the incubation period is contagious, and asymptomatic infected persons may also become the source of infection, which is mainly transmitted through respiratory droplets and cl...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/558G01N33/56983G01N2333/165
Inventor 王胜岚马玉兰韦健华罗俊蓝李敏仪
Owner 中山生物工程有限公司
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