Group of combined markers for diagnosing children asthma and application and detection kit thereof

[0028] 1. Serum Sample Collection

[0029]All volunteers included in the study signed an informed consent form before serum samples were collected. Under the same conditions, blood samples from 22 patients with asthma (0.5-13 years old), 20 patients with bronchiolitis (1-19 months old) and 26 healthy people (0-6 years old) were collected. Serum was taken directly after standing for 60 minutes, and stored in a -80°C refrigerator for future use.

[0030] 2. Analysis method

[0031] 2.1 Serum sample pretreatment

[0032] Add 200 μL methanol containing internal standard (containing 2 internal standards: 1.85 μg / mL D3-cholic acid and 0.75 μg / mL LPC 19:0) to the 96-well protein precipitation plate, then add 50 μL serum, vortex at low speed for 10 min, and then Centrifuge at 400g, discard the upper protein precipitation plate, and freeze-dry the samples in the lower collection box. Before LC-MS analysis, the sample in the collection box was reconstituted with 80 μL of acetonitril...

[0048] 1. Serum Sample Collection

[0049] All volunteers included in the study signed an informed consent form before serum samples were collected. Under the same conditions, blood samples from 11 patients with asthma (0.5-13 years), 10 patients with bronchiolitis (1 month-19 months) and 12 healthy people (0-6 years old) were collected. Serum was taken directly after standing for 60 minutes, and stored in a -80°C refrigerator for future use.

[0050] 2. Analysis method

[0051] Same as Example 1

[0052] 3. Serum test results and auxiliary diagnostic methods

[0053] The relative contents of 2-piperidone and SM 30:1 in normal control group, bronchiolitis group and asthma group are as follows: figure 1 (validation set) and Table 5. Concentrations of 2-piperidone and SM 30:1 were decreased in the serum of asthmatic patients relative to normal controls and bronchiolitis. Table 5. Relative content of 2-piperidone and SM 30:1 in normal controls, bronchiolitis and asthma

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