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
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[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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