Test paper strip using colloidal gold immunochromatographic technology for quantitative determination of serum pepsinogen and preparation method and application thereof
A pepsinogen and quantitative detection technology, which is applied in the field of rapid screening and efficacy evaluation of gastric diseases or early gastric cancer, can solve the problems of long detection time, high detection cost, unsuitable for single test, etc., and achieves the effect of convenient operation.
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Embodiment 1
[0041] Embodiment 1 Preparation of test strips for the quantitative detection of serum pepsinogen using colloidal gold immunochromatography technology of the present invention
[0042] The preparation flow chart is as figure 1 shown.
[0043] 1. Preparation of human pepsin antigen I or protease antigen II natural protein
[0044] Collect surgically resected normal human gastric tissue, rinse with phosphate buffer, separate gastric mucosa, blot dry, weigh, add phosphate buffer, mash and homogenate, centrifuge, collect supernatant, precipitate phosphate buffer weight suspended, centrifuged again, and combined the supernatant to obtain the pepsinogen crude extract;
[0045] Add the pepsinogen crude extract to the equilibrated DEAE-52 chromatographic column, elute the chromatographic column with PBS buffer until the absorbance value is 0 at 280nm, continue to elute the chromatographic column with PBS buffer, and combine the eluted Peak off to obtain active pepsinogen;
[0046]...
Embodiment 2
[0057] The use and result judgment of the colloidal gold test strip of the present invention
[0058] Dilute the prepared human pepsin antigen I and human pepsin antigen II to 22, 46, 63, 100, 200ug / L and 2.8, 4.8, 11.5, 25, 43ug / L solutions respectively, as standard solution, and prepare at the same time blank solution. Take 60μl standard and sample to be tested (30μl sample + 30μl sample diluent) and drop them into the test strip sample groove. After 20 minutes, put the color-developed standard and the test strip of the sample to be tested into the immunochromatography reading In the instrument (Beijing Meikang Biotechnology Research Center Co., Ltd.), read the color value of the T line and record it. The schematic diagram of the color development result is as follows image 3 Shown; Bring the color value of the sample to be tested into the standard curve to obtain the concentration result of pepsinogen I or pepsinogen II.
[0059] The pepsinogen colloidal gold immunochrom...
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