Kit for detecting precursor of gastrin-releasing peptide for facilitating clinical application
A technology of gastrin-releasing peptide and detection kit, which is applied in the field of medical detection, can solve the problems of high price and high cost, achieve low cost of use, avoid detection errors, and facilitate clinical application
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Embodiment 1
[0020] Example 1 Preparation of Gastrin-releasing Peptide Precursor Detection Kit
[0021] 1. Preparation of magnetic particle suspension
[0022] 1.1 Take 30ul of the mixed magnetic particle stock solution and wash it 5 times with 300ul of PBS buffer;
[0023] 1.2 Activate the magnetic particles with 20mg / ml coupling agent for 2 hours;
[0024] 1.3 Wash the activated magnetic particles twice with MES buffer solution of pH 4-5;
[0025] 1.4 Add antibody according to the amount of 0.3ug / test, and coat for 2 hours;
[0026] 1.5 Stop the reaction with 5 mol / L ethanolamine for 1 hour;
[0027] 1.6 Seal with blocking solution for 3 times;
[0028] 1.7 Add 3ml of blocking solution for preservation.
[0029] The blocking solution used is prepared by 0.01M PBS buffer solution of PH7~8, which contains 1~3‰ (w / v) preservative (Proclin 300, MIT, Bro, IPBC or NaN can be used) 3 one or a mixture of two or more of them), 1-3‰ (w / v) protein stabilizer (one or two or more of casein, fis...
Embodiment 2
[0038] Embodiment 2 is used for the sample preservation time of this kit test
[0039]The samples stored at 2~8°C were tested on the 0th day, the 1st day, the 2nd day, the 3rd day, the 5th day, and the 7th day respectively using the kit of the present invention. The values measured on the 3rd day, the 5th day, and the 7th day sample were compared with those on the 0th day, as shown in Table 1 below:
[0040] Table 1
[0041]
[0042] As can be seen from the data in Table 1, the decline rate of the samples in the first 5 days is less than 10%, indicating that the samples used for the test of the kit of the present invention can be stored for 5 days.
Embodiment 3
[0043] Embodiment 3 Performance evaluation of the kit of the present invention
[0044] 1. Sensitivity detection
[0045] LoB, prepare 5 clinical samples with a value close to 0, repeat 3 times for each sample, do 4 days in total, and get 60 data; LoD, prepare 5 serial clinical samples with a concentration range of 1~4 times LOB, each sample Repeat 3 times, do a total of 4 days, and get 60 data; FS: Using the data in the LoD experiment, 5 concentration samples are measured 3 times a day, a total of 4 days, each sample gets 12 results, and the calculation of each sample Mean, SD and CV%, the concentration closest to 20% is the functional sensitivity, the results are shown in Table 2;
[0046] Table 2
[0047]
[0048] It can be seen from the results in Table 2 that the concentration that can be accurately detected in the first batch is 4.125pg / mL, and the concentration that can be accurately detected in the second batch is 4.376pg / mL, both of which meet the requirements. ...
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