Kit for quantitatively detecting NT-proBNP/ST2 by time-resolved fluorescence immunochromatography and application thereof

[0049] 1. The preparation method of the treatment pad:

[0050] The preparation method of the treatment pad comprises:

[0051] The treatment solution is prepared, the treatment solution is soaked into the sample pad by using a paper pad pressing machine, and the soaked sample pad is dried to obtain the treatment pad.

[0052] The specific steps of drying the infiltrated sample pad include: drying the infiltrated sample pad at a temperature of (18-26)°C and a relative humidity of ≤30% for overnight (16-24) hours to obtain a dried Sample pad. Put the dried sample pad in a ziplock bag, seal it in the ziplock bag, and put an appropriate amount of desiccant. Before assembling the intermediate, under the condition of temperature (18~28) ℃ and relative humidity ≤30%, cut off the upper and lower two long sides of the dried sample pad by 5mm, and then cut it into (23±1)mm×(300 ±10) mm in size, obtain the treatment pad and set it aside. Calculate the material balance of the processing

[0085] 3. Example 1: The sample detection limit detection experiment of the kit of this application

[0086]a) NT-proBNP: 5 samples with low concentration of approximate detection limit (5pg / ml) were detected, each sample was tested 5 times, 100uL sample to be tested was added to 300uL sample diluent, after mixing, 100uL was added to Into the sample hole, after 15 minutes, use a fluorescence detector to detect, repeat 25 times. The number of test results that are less than the blank limit (1pg / mL) among the 25 test values ​​is less than or equal to 3 and all 25 test values ​​are not higher than 5pg / mL. The results showed that the detection limit of this kit was not more than 5.00pg / mL.

[0087] Table 5 NT-proBNP detection limit test results

[0088]

[0089]

[0090] b) ST2: Detect 5 low-value samples with an approximate detection limit (3ng / ml), and test each sample 5 times. Take 100uL of the sample to be tested and add 300uL of sample diluent. After mixing, ta

[0093] Example 2: Sample linearity detection experiment of the kit of the present application

[0094] a) Mix the BNP linear reference substance in a certain proportion, the mixed concentrations are 5pg / mL, 100pg / mL, 1000pg / mL, 3000pg / mL, 5000pg / mL, take 100uL of the reference substance and add 300uL of the sample diluent, mix well Take 100uL and add it to the sample hole, repeat the measurement for each reference product 3 times, use a fluorescence detector to detect after 15 minutes, calculate the average value of the feedback concentration (Yi), use Xi as the independent variable, and Yi as the dependent variable to obtain a linear regression Equation, and calculate the linear correlation coefficient r. The results showed that the linear correlation coefficient r was greater than 0.9900.

[0095] Table 7 NT-proBNP linearity test results

[0096]

[0097] b) Mix the ST2 linear reference substance in a certain proportion, the mixed concentrations are 3.125ng / mL,