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Calprotectin heterodimer detection kit and application thereof

A technology of calprotectin and heterodimer, which is applied in the field of medical testing, can solve problems such as quantitative or semi-quantitative detection of calprotectin, achieve huge market value, avoid false negative results, and improve sensitivity and specificity Effect

Pending Publication Date: 2019-12-24
SUZHOU PRECISION BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the kit cannot be used for quantitative or semi-quantitative detection of calprotectin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The assembly of embodiment 1 kit

[0048] 1. Main reagents

[0049] (1) Coating solution: 1.5g Na 2 CO 3 and 2.92g NaHCO 3 Dissolve in 800mL double-distilled water, adjust the pH to 9.6, and dilute to 1000mL;

[0050] (2) Enzyme-labeled secondary antibody: The enzyme-labeled secondary antibody is diluted with HRP protection solution according to the volume ratio of the enzyme-labeled secondary antibody and the protection solution is 1: (2000-10000), and the specification of the enzyme-labeled secondary antibody is 12mL per bottle;

[0051] (3) Positive control substance: use antibody diluent to dilute calprotectin standard substance according to the volume ratio of 1:(20-100), and the specification is 1mL per tube;

[0052] (4) Negative control substance: use antibody diluent to dilute the negative protein according to the volume ratio of 1: (20-100), and the specification is 1 mL per tube;

[0053] (5) Antibody diluent; add 800mL 1×PBS to 1g BSA, and dilute to 100...

Embodiment 2

[0062] ELISA detection of embodiment 2 calprotectin heterodimer

[0063] (1) The kit is equilibrated at room temperature for 0.5-1h, and the calprotectin is diluted with the sample diluent;

[0064] (2) Add 100 μL of the diluted sample to be tested, negative control substance and positive control substance to the corresponding wells of the 96-well ELISA plate, seal the plate, incubate at 37°C for 30 minutes, and remove the supernatant;

[0065] (3) Wash the 96-well plate with washing solution, 250 μL / well, wash 3 times, discard the washing solution, and tap it firmly on absorbent paper to avoid drying the coated well;

[0066] (4) Add 100 μL of biotin-labeled calprotectin monoclonal antibody to the ELISA plate, seal the plate, incubate at 37°C for 60 minutes, wash with washing solution for 3 times, and then tap it firmly on absorbent paper to avoid drying the coated well;

[0067] (5) Add 100 μL of HRP-labeled goat anti-mouse IgG1 to the microtiter plate and incubate at 37°C ...

Embodiment 3

[0080] Embodiment 3 Accuracy experiment

[0081] The kits of Example 1 and Comparative Examples 1-3 were used to measure the calprotectin heterodimer in the feces of 623 healthy donors, and the results are shown in Table 1.

[0082] Table 1 Statistical Table of Clinical Diagnosis Results of the Kit

[0083] Numbering Positive Negative accuracy(%) Example 1 6 617 99.0 Comparative example 1 17 606 97.3 Comparative example 2 45 578 92.8 Comparative example 3 41 582 93.4

[0084] It can be seen that the accuracy of the kit prepared by using the anti-MRP8 / MRP14 antibody as the coating antibody of the present application is higher than that of other kits prepared by using a single antibody as the coating antibody. All are better than the test kit of comparative example.

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Abstract

The invention provides a calprotectin heterodimer detection kit and application thereof. The kit comprises a coating antibody including an anti-myeloid-associated protein 8 / 14 antibody.According to the kit disclosed by the invention, the anti-myeloid-associated protein 8 / 14 antibody is used as a coating antibody and the enrichment effect on the trace amount of calprotectin heterodimer in the sample is realized; and the calprotectin monoclonal antibody is used as the detection antibody to carry out calprotectin detection, so that the interference of other types of calprotectin antigens on the result is basically eliminated and the detection sensitivity and the result accuracy are improved.

Description

technical field [0001] The invention belongs to the technical field of medical examination, and relates to a calprotectin heterodimer detection kit and application thereof. Background technique [0002] Calprotectin (also known as calprotectin) is a heterodimeric protein derived from neutrophils and macrophages, which has the functions of regulating immunity, inducing apoptosis, and inhibiting bacterial growth. Recently, calprotectin Research on the relationship with ulcerative colitis (ulcerative colitis, UC), inflammatory bowel disease (inflammatory bowel disease, IBD) and tumors has gradually become a hot spot. Studies have found that the content of calprotectin in the body fluids and excreta of patients with ulcerative colitis, inflammatory bowel disease and tumors changes, and can be used as a tumor marker for early diagnosis of tumors. [0003] Calprotectin consists of two heavy chains MRP14 and one light chain MRP8 non-covalently combined to form a heterodimer, which...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574G01N33/577
CPCG01N33/6893G01N33/577G01N33/57446G01N2800/065
Inventor 朱浩朱永亮张荃铭穆延召
Owner SUZHOU PRECISION BIOTECH CO LTD
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