tsh immunochromatography kit and preparation method thereof
An immunochromatography and immunochromatographic detection technology, applied in the field of TSH immunochromatography kits and their preparation, can solve the problems of antibody or protein detachment, kit stability and sensitivity cannot be guaranteed, colloidal gold instability, etc. Fast response, simple operation, high sensitivity effect
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Embodiment 1
[0018] TSHα, β subunit-specific paired antibodies were purchased from abcam, USA; other reagents were purchased from sigma.
[0019] The coating process of NC membrane antibody is as follows: TSHα subunit antibody is prepared into a 2mg / ml solution with 0.1M pH8.0 phosphate buffer (PBS), and anti-mouse IgG is prepared into a 1.5mg / ml solution. The film instrument draws lines on the upper and lower parts of the NC film at a parameter of 1.5 μl / cm, and coats the C and T lines. After marking, place the NC film in a drying room at a temperature of 20-25°C and a humidity of less than 30%. Dry for 4 Hour.
[0020] The preparation process of the colloidal gold pad is as follows: a colloidal gold solution with a diameter of 20nm is prepared by the chloroauric acid-trisodium citrate reduction method (Frens method). 2 CO 3 Adjust to pH 7.5, add 2.0mg TSHβ subunit antibody to 100ml colloidal gold solution, then slowly add 0.2M K 2 CO 3 Adjust the colloidal gold solution to pH 9.0, ad...
Embodiment 2
[0023] Embodiment 2 Sensitivity test
[0024]TSH standard products of 0, 0.1, 0.5, 1.0, 2.0, 4.0, and 8.0 μIU / mL were used to measure the test paper card, and each concentration was tested five times in parallel. Detection method: 1) Equilibrate the detection reagent and sample to room temperature, take out the test paper card, and lay it flat; 2) Accurately draw 10 μl of serum or plasma sample, when the sample is whole blood, draw 20ul of sample, add it to the sample hole, and immediately put it in the lower part Add 100 μL sample diluent (PBS) to the buffer well, and within 10-15 minutes, interpret the immunochromatographic results by naked eyes and BiodotTSR3000 reading system. The visual judgment results are judged by the color difference between different concentrations of TSH standard test paper and negative test paper (TSH is 0 μIU / mL), and the instrument judgment results are judged by the OD590 value of different concentrations of TSH standard test paper and negative t...
Embodiment 3
[0028] Embodiment 3 stability test
[0029] After the test paper card prepared in Example 1 was placed at 40°C for 1 month and 3 months respectively, the sensitivity of the test paper was measured according to the method of Example 2.
[0030] The comparative examples of this embodiment are as follows:
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