IL-6 fluorescence immunochromatography kit and preparation method thereof

A fluorescence immunochromatography, IL-6 technology, applied in the field of IL-6 fluorescence immunochromatography kit and preparation thereof, can solve the problems of unsuitability for clinical emergency detection and project promotion, large instrument volume and high reagent cost, It can shorten the detection time, improve the signal-to-noise ratio, and improve the detection sensitivity.

Pending Publication Date: 2022-06-07
上海艾瑞德生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the first two detection products have the problem of low sensitivity
Although the chemiluminescence method has high sensitivity and good accuracy, the cost of reagents is relatively high, the reaction time is long, and the volume of the instrument is large, so it is not suitable for clinical emergency testing and project promotion

Method used

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  • IL-6 fluorescence immunochromatography kit and preparation method thereof
  • IL-6 fluorescence immunochromatography kit and preparation method thereof
  • IL-6 fluorescence immunochromatography kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] This embodiment provides an IL-6 fluorescence immunochromatography kit and a preparation method thereof.

[0048] Specifically include:

[0049] 1. Preparation of fluorescent microsphere-labeled IL-6 monoclonal antibody solution

[0050] (1) Take 100 μL of fluorescent microspheres, add 900 μL of MES buffer (0.05M, pH 6.0), centrifuge at 21000g for 20 min to remove the supernatant, add 1 mL of MES buffer (0.05M, pH 6.0) to resuspend by ultrasonic, and centrifuge at 21000g for 20 minutes to remove the supernatant clear;

[0051] (2) Add 200 μL of MES buffer (0.05M, pH 6.0) for ultrasonic dispersion, and then add 10 μL of NHS (100 mg / mL) and 5 μL of EDC solution (100 mg / mL) to mix well, mix by ultrasonic, and incubate at room temperature on a rotary incubator Reaction 30min;

[0052] (3) After the reaction, 800 μL of MES buffer (0.05M, pH 6.0) was added, and the supernatant was removed by centrifugation at 21,000 g for 20 min;

[0053] (4) Add 1 mL of MES buffer (0.05M...

experiment example 1

[0069] This experimental example provides the assay method of the IL-6 fluorescence immunochromatography kit of Example 1.

[0070] 1. Establishment of standard curve: Dilute the IL-6 standard with the calibrator diluent into a series of gradient concentrations of standard, mix it with the clinical diluent in proportion and add it to the sample hole of the chromatographic test strip. After 10 minutes of reaction, use a dry immunofluorescence analyzer (Shanghai Ai Ruide Biotechnology Co., Ltd., i-Reader S) to read the fluorescence signals of T and C lines on the NC membrane and calculate the ratio of 5000T / (T+C). The concentration of the solution and the ratio were linearly fitted to obtain the standard fitting curve. The results are shown in Table 1 and figure 2 shown.

[0071] Table 1 Calculation of 5000T / (T+C) results

[0072] Concentration (pg / mL) 5000T / (T+C) 0 44.11 2 68.94 7 126.04 10 158.01 70 677.96 140 1150.52 350 1816....

experiment example 2

[0075] In this experimental example, the sensitivity evaluation of the kit was carried out.

[0076] 1. Calculation of blank limit LOB: use the kit of the present invention to repeat the test blank sample 20 times, and calculate the average value of the test results and standard deviation (SD). LOB is The results are shown in Table 2.

[0077] Table 2 LOB calculation results

[0078]

[0079] According to the results in Table 2, the LOB was 0.55 pg / mL.

[0080] 2. Calculation of detection limit LOD: Take 5 low-value samples with approximate LOD for testing, each sample is tested 5 times, and the test results are sorted by size, which should meet: the number of test results below the LOB should not be higher than 3, and the results are shown in Table 3.

[0081] Table 3 LOD calculation results

[0082]

[0083] According to the results in Table 3, after 25 replicates of the sample at a concentration of approximately 1.5 pg / mL, no results below the LOB occurred, th...

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Abstract

The invention discloses an IL-6 fluorescence immunochromatography kit and a preparation method thereof, and relates to the technical field of biological medicines. The kit comprises a sample pad, a combination pad, a buffer pad, a coating film and an absorption pad which are arranged on a bottom plate, the combination pad is provided with an anti-IL-6 monoclonal antibody marked with fluorescent latex microspheres and a DNP-BSA antibody marked with fluorescent latex microspheres, the coating film is provided with a detection T line and a quality control C line, the detection T line is coated with the anti-IL-6 monoclonal antibody, the quality control C line is coated with the DNP-BSA antibody, and the quality control C line is coated with the DNP-BSA antibody. And the quality control C line is coated with an X anti-DNP polyclonal antibody. The kit can improve the detection sensitivity of IL-6, improve the detection accuracy and shorten the detection time.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an IL-6 fluorescence immunochromatography kit and a preparation method thereof. Background technique [0002] Interleukin-6 (IL-6) is a cytokine with various biological activities. It can regulate the growth and differentiation of various cells, regulate immune response, hematopoietic function, neuroendocrine system, etc., and play a role in the body's anti-infection important role in the immune response. It is involved in the occurrence and development of many diseases, and viral or bacterial infections, autoimmune diseases, etc. can lead to elevated serum levels. And when infection and inflammation occur, IL-6 can peak within 1-2 hours, while procalcitonin (PCT) increases after 2 hours, and C-reactive protein (CRP) increases after 6 hours. Therefore, IL-6 is a sensitive indicator for the early diagnosis of acute infection, especially for the differential diagnosis of earl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/542G01N33/543G01N33/577
CPCG01N33/558G01N33/542G01N33/543G01N33/577
Inventor 王茜倪海欧陈超樊菲于坤宏
Owner 上海艾瑞德生物科技有限公司
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