IgM antibody joint detection device and method for coxsackievirus A16 and enterovirus 71
A technology for coxsackie virus and enterovirus, which is applied in measurement devices, disease diagnosis, biological testing, etc., can solve the problems of high-level structure impact of amplified fragments, high RT-PCR requirements, and difficulty in finding universal primers. Novel, improved sensitivity, simple structure effect
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[0033] The preparation method of the IgM antibody combined detection device of coxsackievirus A16 and enterovirus 71 of the present invention comprises the high-specificity coxsackievirus A16 (CA16) antigen and enterovirus 71 (EV71) antigen of solid phase purification ) antigen (detection line T1, T2) and goat anti-mouse IgM polyclonal antibody (control line) of nitrocellulose membrane, glass fiber adsorbed with colloidal gold-labeled mouse anti-human IgM, sample pad, absorbent paper and other auxiliary materials are thus pasted production. Specific steps are as follows:
[0034] (a) Preparation of colloidal gold by trisodium citrate reduction method;
[0035] (b) using colloidal gold labeled mouse anti-human IgM prepared in step (a) to obtain immune colloidal gold;
[0036] (c) Diluting the immune colloidal gold in step (b) with a gold spraying buffer to obtain an immune colloidal gold solution, and spraying the immune colloidal gold solution on the glass fiber mat to prepa...
Embodiment
[0044] Coxsackie virus A16 (CA16) and enterovirus 71 (EV71) IgM antibody combined detection device
[0045] 1. Preparation of colloidal gold solution
[0046] Quickly add gold chloride into heated 100ml purified water, and after the solution boils again, quickly add trisodium citrate, gold chloride: trisodium citrate 1: (0.5-2), continue to boil, and observe that the color of the solution changes from yellow to After the black turns purple again, and finally turns into a stable wine red, continue heating for 10 minutes by timing. The colloidal gold particles are 20-60nm. In this embodiment, 30nm colloidal gold is selected for the next experiment.
[0047] 2. Preparation of immunocolloidal gold
[0048] 1) Take 100ml of 30nm colloidal gold solution, add 140μl of pH regulator, mix well; let stand for 5min;
[0049] 2) In the 30nm colloidal gold solution, add mouse anti-human IgM at a ratio of 14 μg per milliliter of colloidal gold solution, a total of 1.4 mg, mix well; let st...
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