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Improved gastrin 17 detection kit
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A detection kit and technology of gastrin, applied in the field of immunological detection, can solve the problems of inconvenient clinical use and achieve the effects of simple operation, elimination of differences between old and new samples, and improvement of detection accuracy
Inactive Publication Date: 2019-02-05
AUTOBIO DIAGNOSTICS CO LTD
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These problems have brought great inconvenience to the clinical use of gastrin 17 project
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Embodiment 1
[0022] Example 1 Preparation of PBS buffer solution (according to 1000 mL volume)
[0023] Weigh 0.592g NaH 2 PO 4 .2H 2 O, 5.80 g Na 2 HPO 4 .12H 2 Add O and 9.0g NaCl to 1000mL purified water, add HCl to adjust the pH to 7.4±0.05, add preservatives P300 1g, MIT 1g, mix well, store at 2-8°C until use.
Embodiment 2
[0024] Example 2 Preparation of Improved Gastrin 17 Detection Kit
[0026] First prepare blocking solution (according to 1000 mL): take 100 mL of PBS buffer, add 2.5% bovine serum albumin (BSA), mix well, aliquot, and store at 2-8°C until use.
[0027] Fully mix 30ul of the magnetic particle stock solution, wash 3 times with 300ul of phosphate buffer, add 100ul of coupling activator, mix and shake for 2 hours, then add G-17 antibody at a concentration of 0.1 ug / T, mix and shake for 2 hours h, and finally blocked four times with blocking solution, dilute to 3ml, and store at 2-8°C until use.
[0029] Add HRP-labeled G-17 antibody to PBS buffer containing 2% BSA at a volume ratio of 1:1000, then mix evenly, aliquot, and store at 2-8°C until use.
[0034] Embodiment 3 Detection method of the kit of the present invention
[0035]Add calibrators and samples to the reaction container at one time, and the sample volume is 25ul / well. Add 20ul of magnetic particle suspension and 50ul of sample diluent to each well, mix well and incubate at 37°C for 15min, wash with washing solution 5 times. Add 100ul of enzyme conjugate to each well, mix well and incubate at 37°C for 17min, wash with washing solution 5 times, add 50ul each of substrate A solution and substrate B solution to each well, and detect the luminescence value within 1-5min after mixing. The concentration value of G-17 was calculated back through the calibrator curve.
[0036] The kit of the present invention plays the role of eliminating the difference between old and new samples by mixing the EDTA in the sample diluent with the sample.
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Abstract
The invention discloses an improved gastrin 17 detection kit. The detection kit comprises a G-17 monoclonal-antibody-coated magnetic particle suspension, an HRP-marked G-17 monoclonal-antibody-containing enzyme conjugate, a sample diluent and a calibration product, wherein the sample diluent is prepared from 0.01-0.2mol / L of buffer solution, 0.1-0.5%(w / v) of preservative, 1-10%(w / v) of protectiveprotein, 1-10%(w / v) of EDTA, 0.1-0.5%(w / v) of sodiumdodecyl sulfate, 0-500[mu]g / mL of blocking agent and 0.001%(g / v) of pigment. The kit is prepared based on a magnetic particle chemiluminescence method, and is matched with an Auto Lumo A2000 series full automatic immunoassay instrument for use, so that the automatic detection can be realized, the operation is simple and convenient, and the detection speed is high. The kit adopts a unique sample diluent, so that the difference of new and old samples can be eliminated, and the false positive or missing detection probability caused by HAMA is effectively reduced, thereby highly improving the detection accuracy of the gastrin 17.
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