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Interleukin 6 chemiluminiscence detection kit and preparation method thereof

A chemiluminescence detection and chemiluminescence technology, which is applied in the fields of chemiluminescence/bioluminescence, biological testing, and analysis through chemical reaction of materials, etc. The chromogenic substrate is easy to decompose when exposed to light, and the test results are not accurate, etc., to achieve the effect of fast and convenient detection, stable luminescence value, and high accuracy

Inactive Publication Date: 2018-03-20
太原瑞盛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of using ELISA for detection is that horseradish peroxidase or alkaline phosphate markers are easily inactivated, the chromogenic substrate is easily decomposed when exposed to light, and the sensitivity is low. inaccurate result

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: The formation and preparation of kit

[0029] 1) Kit set-up

[0030] A chemiluminescent detection kit for interleukin-6 is formed, which contains the following components:

[0031] sample diluent;

[0032] Magnetic particles coated with interleukin-6 monoclonal antibody;

[0033] Interleukin-6 monoclonal antibody labeled with a chemiluminescent marker;

[0034] Excitation solution A, excitation solution B;

[0035] Interleukin 6 calibrator solution, the concentrations are 0pg / mL, 10pg / mL, 50pg / mL, 150pg / mL, 600pg / mL, 1000pg / mL;

[0036] Cleaning fluid.

[0037] 2) Preparation of sample diluent

[0038] Specifically, it is a phosphate buffered saline solution with a pH of 7.4, containing 1% bovine serum albumin, 0.1% TritonX-100 and 0.3% preservative Proclin.

[0039] 3) Preparation of magnetic particle suspension coated with interleukin-6 monoclonal antibody

[0040] After magnetically separating the carboxylated magnetic particle solution, remove ...

Embodiment 2

[0051] Embodiment 2: Detection of interleukin-6 in actual samples

[0052] The operating procedures for the use of the interleukin-6 quantitative detection kit of the present invention are as follows:

[0053] Detection of the kit

[0054] 1) Add 50 μL of the sample to be tested, 150 μL of the magnetic particle suspension, and 150 μL of the acridinium ester marker into the reaction tube in sequence, shake and mix well, and incubate at 37°C for 15 min.

[0055] 2) Separate and wash 5 times.

[0056] 3) Fully shake the washed reaction container to disperse the magnetic particles.

[0057] 4) Add 100 μL of chemiluminescence excitation solution A, and add 100 μL of chemiluminescence excitation solution B after 1 s, and measure its relative luminous intensity. The content of procalcitonin in the sample is proportional to its relative luminous intensity.

Embodiment 3

[0058] Embodiment 3: the performance index of kit

[0059] 1) Sensitivity of the kit

[0060] Use the 0 ng / mL calibrator in the kit as the sample to be tested, repeat the measurement 20 times, and obtain the RLU value (relative luminescence value) of the 20 measurement results, calculate the mean (M) and standard deviation (SD), and get Get the RLU value corresponding to M+2SD, and perform a two-point regression fitting based on the concentration-RLU value between the zero concentration calibrator and the adjacent calibrator to obtain a linear equation, and bring the RLU value of M+2SD into the above equation , find the corresponding concentration value. According to the detection method of the lowest detection limit in the detection scheme, the experiment was repeated 3 times, and finally the analytical sensitivity of the kit for interleukin-6 was found to be 3.8 pg / mL.

[0061] 2) Precision determination

[0062] The samples with the concentration of 50pg / mL and 600pg / mL ...

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Abstract

The invention discloses a chemiluminescent detection kit for interleukin-6 and a preparation method thereof. The kit includes: sample diluent, magnetic particles coated with interleukin-6 monoclonal antibody, interleukin-6 monoclonal antibody labeled with acridinium ester, interleukin-6 series standard solution, chemiluminescence excitation solution A, chemiluminescence excitation solution B. Cleaning solution. The kit of the invention combines the chemiluminescent technology with the immunomagnetic particle, and provides a nearly homogeneous reaction system. Compared with the prior art, the kit of the invention has the advantages of high sensitivity, strong specificity, short reaction time and the like.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis, in particular to a kit for detecting interleukin-6 in human serum by a chemiluminescence method and a preparation method thereof. Background technique [0002] Interleukin 6, the full name of interleukin-6 (IL-6), is a multi-effect cytokine, which belongs to a kind of interleukin. It is produced by fibroblasts, monocytes / macrophages, T lymphocytes, B lymphocytes, epithelial cells, keratinocytes, and various tumor cells. IL-6 can be induced by IL-1, TNF-a, PDGF, virus infection, double-stranded RNA and cAMP, etc. to induce normal cells to produce. Interleukin 6 can stimulate the proliferation, differentiation and function of cells involved in the immune response. The main function of interleukin 6 is: promote the proliferation and differentiation of B cells and secrete antibodies, and have extensive effects on liver cells, T cells, nerve tissues, and hematopoietic system; have anti-tumor effect...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N33/532G01N21/76
CPCG01N33/6869G01N21/76G01N33/532G01N33/54326G01N33/577
Inventor 李瑶许殊荣程月萍杜爱铭徐兵
Owner 太原瑞盛生物科技有限公司
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