Tumor marker colloidal gold immunochromatographic assay quantitative detection test paper and method for preparing same
A technology of tumor markers and immunochromatography, applied in the field of colloidal gold immunochromatography quantitative detection test strips and its preparation, to achieve short detection cycle, high detection sensitivity and good specificity
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Embodiment 1
[0028] The preparation method of test strip and test paper box of the present embodiment comprises the following steps:
[0029] 1. Antibody preparation
[0030] Use commercially available paired mouse anti-human free prostate-specific antigen (fPSA) monoclonal antibodies Ⅰ and Ⅱ (product model: A8014-1, A8014-2 Beijing Bomai Century Biotechnology Co., Ltd.), 20mM, pH7.4 Dialyze against PBS at 4°C overnight.
[0031] 2. Coating of nitrocellulose membrane
[0032] Coating buffer preparation: 0.02M phosphate buffer solution (PBS) with pH 7.4 was used as the coating buffer, which was sterilized by 0.22um microporous membrane and stored at 4°C for later use, valid for two weeks.
[0033] Preparation of blocking solution: 0.02M phosphate buffer saline (PBS) containing 0.5% BSA, pH 7.4, filtered and sterilized by a 0.22um microporous membrane, and stored at 4°C for later use, valid for one week.
[0034] Dilute the mouse anti-human free prostate specific antigen monoclonal antibo...
Embodiment 2
[0045] The usage method of the tumor marker rapid quantitative detection test paper of the present invention:
[0046] Mix the sample to be tested with the sample diluent, then insert the test paper into the sample mixture, the liquid in the sample moves upwards by siphon action, and the result is interpreted within 10-15 minutes. Quantification of tumor markers can be performed according to the following two methods:
[0047] 1. Visual detection: count the number of color T-lines and observe the color depth, compare the color development table of the standard solution, and read the concentration of free PSA from the table.
[0048] Prepare free-state prostate-specific antigen colloidal gold immunochromatographic test strips according to the determined optimal conditions, and detect free-state prostate-specific antigen at different concentrations. The sample treatment solution for free-state prostate-specific antigen was prepared into 1ng / ml, 3ng / ml, 5ng / ml, 7ng / ml. Three re...
Embodiment 3
[0052] Stability test The newly prepared colloidal gold immunochromatographic test strips were stored at 37°C for 8 days, and tested with 1ng / ml, 3ng / ml, 5ng / ml, and 7ng / ml tumor marker free prostate specific antigen respectively. It shows that compared with the newly prepared detection test strip, the sensitivity is not significantly decreased, and the specificity is good.
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