A quick test method for ELISA with microwave as medium
An enzyme-linked immunosorbent and microwave technology, applied in the field of rapid assay, can solve the problems of non-specific binding and material death
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example 1
[0106] Carrier activation
[0107]The holes of the module (12-hole polystyrene module, Dynatech, USA) were loaded on 1.82 mg of 1-fluoro-2-nitro-azidobenzene (1-fluoro-2-nitro-azidobenzene) (FNAB), dissolved Dry appropriately in 100 μl of methanol per well in the dark. Then in a UVStrtalinker2400 (Stratagen , USA) with UV light at a wavelength of 365 nm to irradiate FNBA-coated apertures for 10 minutes or in bright sunlight for 1 hour. The wells were then rinsed several times with methanol to remove unbound adapters and dried at room temperature. These activated pores of the module are used to immobilize antigens or antibodies in the process of the invention.
example 2
[0109] Immobilization of Entamoeba Histolytica Antigen by Microwave Irradiation
[0110] E. Histolytica antigen (1 μg) diluted in 100 μl of PBS was loaded in an activated hole of a module and subjected to 10 seconds in a microwave oven (BPL-Sanyo, India) microwave radiation, the operating frequency of the microwave oven is about 2450MHz, and the maximum output power is 700 watts. Irradiation was performed in a microwave oven with a maximum power setting of 10, that is, an output power of 700 watts at 100% magnetron duty cycle.
[0111] The wells were rinsed extensively with wash buffer to remove unbound antigen. Subsequent steps were carried out using conventional procedures. Therefore, blocking with blocking solution (200 μl), binding of antibody (100 μl) and binding of anti-rabbit immunoglobulin G (IgG)-horseradish peroxidase conjugate (100 μl) were performed at 37°C It is carried out by incubation, and each step lasts 2 hours. Thoroughly flush the hole with flushing flu...
example 3
[0116] Plugging free surfaces with plugging agents under microwave irradiation
[0117] As described above, E. Histolytica antigen was immobilized in an activated hole of a module by microwaves. After extensive rinsing with rinsing solution, 200 [mu]l of rinsing solution was added to the well and irradiated with a microwave at 700 watts for 10 seconds. Subsequent antibody and conjugate binding steps were performed as described in Example 2 outside of the microwave oven. Color development and absorbance readings were also performed as described in Example 2.
[0118] Similar experiments were performed with negative sera.
[0119] In order to verify the optimum time for plugging, two different experiments were carried out in the same manner except for the microwave exposure time, which was increased to 40 and 60 seconds. All experiments were performed in triplicate wells.
[0120] The optimization results of plugging time under microwave irradiation are shown in Table 2.
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