Application of gelsolin detection substance in preparation of uterine cancer evaluation and detection reagent
A gelsolin and detection reagent technology, applied in the biological field, can solve the problems of no relevant reports on uterine cancer and a survival rate of less than 30%.
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Embodiment 1
[0025] Preparation of gelsolin detection reagent (time-resolved fluorescence method)
[0026]The detection card in the gelsolin detection reagent consists of a sample pad, a fluorescent pad, a detection pad with a detection line and a quality control line, a sample suction pad and a bottom plate. Its specific preparation method is as follows:
[0027] 1. Fluorescent microsphere-labeled antibody (take 500 copies / batch as an example)
[0028] 1) Take 250 μL of fluorescent microsphere solution and centrifuge;
[0029] 2) Add 5 mg of imine, shake at room temperature for 30 min in the dark, and discard the supernatant by centrifugation;
[0030] 3) Wash with 0.01M PBS, pH 7.4;
[0031] 4) Add 100 μL of 0.5 mg / mL GSN-labeled antibody;
[0032] 5) 1% BSA was added, and the reaction was shaken at room temperature for 30 min in the dark.
[0033] 2. Spray the marked solution on the fluorescent pad, and dry it in a drying oven for 1 hour at room temperature.
[0034] 3. Antibody c...
Embodiment 2
[0037] Use of gelsolin detection reagent: Aspirate the sample to be tested at room temperature (serum / plasma 5μL or whole blood 10μL), add it to 1000μL of sample diluent, and after mixing, pipette 80μL vertically into the sample hole, and let stand for 10 minutes, read the data with a fluorescence immunoassay analyzer.
Embodiment 3
[0039] The relationship between gelsolin level and uterine cancer
[0040] 1. Sample
[0041] Serum samples were collected from healthy people in the physical examination center, ranging in age from 40 to 70 years old; among them, 300 patients with more than 3 risk factors for uterine cancer were regarded as the risk group; there were no risk factors for uterine cancer and no symptoms or evidence related to uterine cancer. As the control group, 200 cases; and 150 serum samples from patients who were clinically diagnosed with uterine cancer but had not received treatment were used as the uterine cancer group.
[0042] 2. Serum Preparation
[0043] 5ml of venous blood was drawn into a centrifuge tube (without anticoagulant), left standing at room temperature for 30 minutes, centrifuged (2000rpm / min), and the upper serum was drawn into aliquots, 100μl per tube, and stored in a -70°C refrigerator.
[0044] 3. Experimental method
[0045] The GSN content in the three groups of s...
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Abstract
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