A composition, chip and preparation method thereof, and detection device comprising the chip
一种组合物、芯片的技术，应用在测量装置、生物材料分析、仪器等方向，能够解决自身免疫性肾病检测等问题，达到储存及运输条件便利、稳定性好、降低成本的效果
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Embodiment 1～3
[0075] 1. The coating of the chip:
[0076] 1) The chip array design and the specific distribution of antigens and reference points are shown in Table 1 and Table 2:
[0077] Table 1
[0078] · · · · · · · · · · · · · · · · · · · ·
[0079] Table 2
[0080] PC NC GBM LAMP-2 Endothelial cells SC S1 PR3 PLA2R nucleosome EC S2 MPO THSD7A dsDNA Loc S3 LF PT protein C1q
[0081] 2) The specific coating process:
[0082] First, dilute antibodies and related proteins as follows:
[0083] The PC, NC, S1, S2, S3, and EC spots in the array were coated with 2μg / ml, 0.01μg / ml, 0.5μg / ml, 2μg / ml, 4μg / ml, 2μg / ml human IgG, respectively, diluted The buffer was CB buffer at pH 9.6 (containing 2.5% PEG4000, 5% trehalose, 0.05% Proclin300, and 15% glycerol).
[0084] The SC spots were coated with 2 μg / ml goat anti-human IgG antibody, and the dilution buffer was CB buffer at pH 9.6 (same as abo...
Embodiment 4
[0107] Example 4 Accuracy Evaluation
[0108] 1. 10 cases of anti-GBM IgG positive serum and 10 cases of anti-GBM IgG negative serum screened by ELISA kit of Abnova company were tested with the chip kit prepared in Examples 1 to 3. The results are as follows in Table 5 (+ means positive, - means negative).
[0109] Table 5 Chip test anti-GBM IgG serum results prepared in Examples 1-3
[0110]
[0111] 2. 10 cases of anti-PR3IgG positive serum and 10 cases of anti-PR3IgG negative serum were screened with the ELISA kit of EUROIMMUN company, and tested with the chips prepared in Examples 1 to 3. The results are as follows in Table 6 (+ means positive, - means negative).
[0112] Table 6 Anti-torch-IgG antibody spectrum chip test results of anti-PR3IgG serum prepared in Examples 1-3
[0113]
[0114] 3. 10 cases of anti-MPO IgG positive serum and 10 cases of anti-MPO IgG negative serum were screened by ELISA kit from Invitrogen Company, and the chips prepared in Examples 1...
Embodiment 5
[0145] Example 5 Sensitivity and specificity test results
[0146] The patient samples were 85 positive serum samples from patients with autoimmune nephropathy diagnosed by renal biopsy and clinical manifestations, and 102 negative serum samples from normal healthy people.
[0147] Table 17
[0148]
[0149] It can be seen from the above data that the detection sensitivity (positive coincidence rate) of the chip kit of the present invention for clinically autoimmune nephropathy patients (autoimmune nephropathy patients diagnosed jointly by renal puncture and clinical symptoms) can reach 98.8%, and the specificity can reach 98.8%. (negative correct rate) reaches 100%, indicating that the kit of the present invention has high sensitivity and good specificity, and can provide more accurate and comprehensive reference and guidance for the diagnosis of autoimmune nephropathy.
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