Colloidal gold kit for jointly detecting coronavirus IgM/IgG antibody, and preparation method thereof

A coronavirus, joint detection technology, applied in measuring devices, instruments, scientific instruments, etc., to achieve the effects of easy product use, low cost, and simple preparation process

Active Publication Date: 2020-05-01
中山生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Since the development of the epidemic, 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

Method used

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  • Colloidal gold kit for jointly detecting coronavirus IgM/IgG antibody, and preparation method thereof
  • Colloidal gold kit for jointly detecting coronavirus IgM/IgG antibody, and preparation method thereof

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Experimental program
Comparison scheme
Effect test

preparation example Construction

[0039] 1. Screening of preparation conditions for gold-labeled N protein

[0040] 1.1. Determination of the particle size of colloidal gold

[0041] 1.1 Materials and methods

[0042] 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.

[0043] Test method: Take 2mL of 1wt% chloroauric acid (HAuCl4) solution, add it to 100mL of water, heat to boil, then add 1-8mL of 1wt% trisodium citrate, mix and boil for 5 minutes until the color does not change.

[0044] Experiment on the labeling effect of colloidal gold particles with different particle sizes: Gold particles with different diameters were prepared by the above method, and the mixed antibody containing 6×His-labeled nucleocapsid protein of the new coronavirus was made in...

Embodiment 1

[0088] Embodiment 1, the preparation method of the colloidal gold kit that jointly detects novel coronavirus IgM / IgG antibody:

[0089] S1: Production of gold conjugate: 3: Take 2mL of 1wt% chloroauric acid and add it to 100mL of aqueous solution, heat to boiling, then add 2mL of 1wt% 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 7.4, add 20μg of the new coronavirus nucleocapsid protein containing 6×His tag and mix well Then add 100 μL of 10vol% bovine serum albumin and mix well to obtain the gold standard solution; centrifuge the gold standard solution, discard the supernatant, add the gold standard complex solution (0.01MPB+1wt%BSA) containing 20wt% sucrose to dissolve, and obtain the gold standard solution. Label N protein; apply gold-labeled N protei...

Embodiment 2

[0092] Embodiment 2, the preparation method of the colloidal gold kit of joint detection novel coronavirus IgM / IgG antibody:

[0093] S1. Preparation of gold conjugate / 3: Take 2mL of 1wt% chloroauric acid and add it to 100mL of aqueous solution, heat to boiling, then add 2.5mL of 1wt% trisodium citrate and mix well, boil until the color of the colloidal gold solution changes from blue to purple to purple , cooled to obtain colloidal gold; take 1 mL of colloidal gold and add 100 μL of 0.1M, pH7.2 PB, mix well, add 0.1M NaOH solution to adjust the pH value to 7.2, add 25 μg of the new coronavirus nucleocapsid protein containing 6×His markers to fully Mix well, then add 100 μL of 10vol% bovine serum albumin and mix to obtain the gold standard solution; centrifuge the gold standard solution, discard the supernatant, add gold standard complex solution (0.01MPB+1wt%BSA) containing 20wt% sucrose to dissolve, and obtain Gold-labeled N protein; apply the gold-labeled N protein on the g...

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Abstract

The invention discloses a colloidal gold kit for jointly detecting coronavirus IgM/IgG antibody, and a preparation method thereof, and relates to the field of biological medicine. Whether anti-novel coronavirus nucleocapsid protein IgM antibody and/or anti-novel coronavirus nucleocapsid protein IgG antibody exists in human serum or plasma or not by adopting an antigen-antibody sandwich method anda colloidal gold immunochromatography method principle, the novel coronavirus nucleocapsid protein containing 6xHis mark is marked by applying colloidal gold, thereby forming gold-marked N protein tobe adsorbed on a gold-marked pad, the novel coronavirus nucleocapsid protein containing 6xHis mark is used as an indication marker, the mouse-anti-human u chain monoclonal antibody is coated on the IgM detection line of a NC membrane, the mouse-anti-human IgG monoclonal antibody is coated on the IgG detection line and the mouse-anti 6xHis monoclonal antibody is coated on a quality control line ofthe NC membrane, the qualitative detection of the anti-novel coronavirus nucleocapsid protein IgG antibody is realized, and the colloidal gold kit disclosed by the invention has the advantages of being convenient to use, high in sensitivity 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 joint detection of novel coronavirus IgM / IgG antibodies and a preparation method thereof, in particular to a colloidal gold immunochromatography method applying the principle of a sandwich method to realize joint detection of anti-antibodies in blood. A kit for the qualitative detection of the IgM antibody to the nucleocapsid protein of the novel coronavirus and / or the IgG antibody against the nucleocapsid protein of the novel coronavirus. Background technique [0002] Since December 2019, there has been an outbreak of a novel coronavirus (2019‑nCoV). The common signs include 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 contagiou...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/532
CPCG01N33/532G01N33/558G01N33/56983
Inventor 王胜岚马玉兰吴伟铭肖慧芳范洁云
Owner 中山生物工程有限公司
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