Prepared carrier for detecting specific antibodies of allergen and its prepn
An allergen-specific technology, which is applied in the field of detection of multiple allergen-specific antibody preparation carriers, can solve the problems of cumbersome operation, great pain for patients, and inability to detect multiple allergen-specific antibodies at the same time, achieving simple operation, pain less effect
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Embodiment 1
[0010] The allergens were prepared with PH7.2 PBS buffer solution to a concentration of 2-50ug / ml, respectively, and were spotted on 10×10cm cellulose acetate membranes with a sample spotter, and recorded. Each allergen was sampled twice, the distance between the repeated points was 100um, and the distance between the two allergens was 150um. The first two dots of the positive control in the first horizontal row, the last two dots of the negative control in the last horizontal row, each time the sample volume is 10nl.
[0011] All allergen points, positive control points and negative control points form a rectangular combination. A total of 90 identical combinations were spotted on a 10×10 cm cellulose acetate membrane. Place the spot film at 37°C for 20-60 minutes, take it out, seal it with 2% BSA routinely, stick it on a double-sided tape after drying, seal it, and store it at 2-8°C.
[0012] When in use, according to the number of detected samples, cut out the combination...
Embodiment 2
[0014] Take a prepared preparatory carrier of the present invention, stick it on a glass slide, add 100 ul of the serum sample to be tested, make it cover all the allergen combination points, put it at 37°C for 1 hour, wash it with washing solution 5 times, each time 5 Add horseradish peroxidase-labeled anti-human IgE (use anti-human IgG when measuring IgG; use anti-human IgA when measuring IgA) 100ul, make it cover all combination points, wash 5 times at 37°C for 1 hour, Each time is 5 minutes, add 50ul luminescence substrate, react for 30-60 seconds, and detect on the luminescence scanner. Those with obvious luminous spots are positive for the allergen-specific IgE (or IgG, IgA). Those without luminous points are negative.
Embodiment 3
[0016] Quantitative determination of allergen-specific IgE (or IgG or IgA)
[0017] The preparatory carrier prepared in Example 1 is used in conjunction with the preparatory carrier prepared by the same method with only anti-human IgE (or anti-human IgG or IgA) and corresponding markers, substrates, etc., to simultaneously quantitatively detect all allergies on the preparatory carrier. Original specific IgE (or IgG or IgA).
[0018] Get the carrier combination prepared in Example 1 and 6 preparations prepared by the same method and only have anti-human IgE (or IgG or IgA) repeated 3 points at a time, and paste them on the same glass slide at intervals, and add 100ul of the serum sample to be tested for the allergen preparation carrier, The whole combination should be covered, add 0, 0.5, 1, 1.5, 25, 100 Iu / ml standard IgE (or IgG or IgA) to the anti-human IgE (or IgG or IgA) preparation carrier respectively, set at 37°C for 1 hour, and wash 5 times 5 minutes each time, add ho...
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