Pepsinogen I/pepsinogen II detection kit
A technology of pepsinogen and detection kits, applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve good detection specificity, ensure accuracy, accurate and reliable diagnosis
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Embodiment 1
[0026] Such as figure 1 As shown, the pepsinogen I / pepsinogen II detection kit is a detection kit for fluorescence quantitative immunochromatography, including a detection card, and the detection card is sequentially provided with: PVC plate 1, sample pad 2, Combination pad 3, chromatographic membrane 4 and absorbent paper 5; wherein the chromatographic membrane includes a detection area and a quality control area; the detection area is coated with a detection area of anti-PG I monoclonal antibody and PG II monoclonal antibody, for capturing antigen; The quality control area is coated with goat anti-mouse IgG antibody for signal detection. Above-mentioned chromatographic membrane 4 is nitrocellulose membrane, makes through the following steps:
[0027] 1) Using a cell line different from the anti-PG I and anti-PG II monoclonal antibody cell lines used on the binding pad 3, prepare and purify anti-human cardiac troponin I antibody and anti-myoglobin antibody by standard asci...
Embodiment 2
[0079] In order to improve the stability and sensitivity of the kit, the optimization measures taken also include:
[0080] The rare earth fluorescent microsphere-labeled anti-PG I monoclonal antibody and PG II monoclonal antibody-microsphere coupling complex adsorbed on the binding pad 3 were prepared by the following steps:
[0081] The aldylation step of rare earth fluorescent microspheres is as follows: take 5 mg of rare earth fluorescent microspheres, wash with 20 mM carbonate buffer solution with pH 9.5 for 3 times by centrifugation at 12000 rpm for 5 minutes, and finally resuspend Add 500ul of aldylated dextran and 0.2μg of 2-aminoethanesulfonic acid to 100ul of the above-mentioned carbonate buffer solution containing 1.5ul of thioglycolic acid monoethanolamine, mix well, and react in the dark at room temperature for 4 hours. Wash by centrifugation and resuspend in 100ul of the above-mentioned carbonate buffer, and place at 4°C for later use. The presence of monoethano...
Embodiment 3
[0083] Stability test
[0084] Stability of the test card: The test card in the kit is usually stored in a dry and cool environment at room temperature. The test card is placed in an oven at 50°C for one month to conduct an accelerated damage stability test, which is equivalent to the validity of the test card stored at room temperature for one year. Place the test card in an oven at 50°C for 1 week, 2 weeks, 3 weeks, and 4 weeks, then take it out and put it in a drying room, and compare it with the reagent card placed in the drying room at room temperature. card stability. The detection card of Example 2 was placed in an oven at 50°C for accelerated destruction for 1 week, 2 weeks, 3 weeks, and 4 weeks. The reaction curve in the case of the standard product test was basically consistent with the detection card stored at room temperature, and the fluorescence signal of low, medium and high concentrations The T / C value basically did not change, indicating that the fluorescenc...
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