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Chemluminescent detection kit for procalcitonin and preparation method of chemluminescent detection kit

A technology for chemiluminescence detection and procalcitonin, which is used in measurement devices, scientific instruments, instruments, etc., can solve the problems of low sensitivity, narrow linear range, and difficulty in full automation of enzyme-linked immunosorbent assay, and achieve fast and convenient detection. Simple and convenient operation, high accuracy

Inactive Publication Date: 2018-03-16
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

Enzyme-linked immunoassay has methodological limitations such as low sensitivity, narrow linear range, and difficulty in realizing full automation.

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 chemiluminescence detection kit for procalcitonin is formed, which contains the following components:

[0031] sample diluent;

[0032] Magnetic particles coated with monoclonal antibody to procalcitonin;

[0033] Procalcitonin monoclonal antibody labeled with a chemiluminescent marker;

[0034] Excitation solution A, excitation solution B;

[0035] Calcitonin original calibration solution, the concentrations are: 0ng / mL, 0.04ng / mL, 0.4ng / mL, 4ng / mL, 10ng / mL, 50ng / mL, 100 ng / 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 procalcitonin monoclonal antibody

[0040] After magnetically separating the carboxylated magnetic parti...

Embodiment 2

[0050] Example 2: Detection of procalcitonin in actual samples

[0051] The procedure for using the procalcitonin quantitative detection kit of the present invention is as follows:

[0052] Detection of the kit

[0053] 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.

[0054] 2) Separate and wash 5 times.

[0055] 3) Fully shake the washed reaction container to disperse the magnetic particles.

[0056] 4) Add 100 μL of chemiluminescence excitation solution A, and add 100 μL of chemiluminescence excitation solution B after 1 s to measure the relative luminous intensity. The content of procalcitonin in the sample is proportional to its relative luminous intensity.

Embodiment 3

[0057] Embodiment 3: the performance index of kit

[0058] 1) Sensitivity of the kit

[0059] Use the 0 ng / mL calibrator in the kit as the sample to be tested, repeat the measurement 20 times, 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 two-point regression fitting according to the concentration-RLU value results 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 procalcitonin was found to be 0.02 ng / mL.

[0060] 2) Precision determination

[0061] The samples with concentrations of 4 ng / mL and 50 ng...

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Abstract

The invention discloses a chemiluminescent detection kit for procalcitonin and a preparation method thereof. The kit includes: sample diluent, magnetic particles coated with procalcitonin monoclonal antibody, procalcitonin monoclonal antibody labeled with acridinium ester, procalcitonin 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 procalcitonin in human serum by a chemiluminescence method and a preparation method thereof. Background technique [0002] Procalcitonin (PCT) is a glycoprotein composed of 116 amino acids and a molecular weight of 13kDa. It is the precursor of serum calcitonin (CT). Its half-life in the human body is about 20-24 hours, and it has good stability. . Procalcitonin is the expression product of calcitonin I gene (CALCI) on human chromosome 11. The gene is a single-copy gene, consisting of 2800 base pairs, containing 6 exons and 5 introns, and the molecular weight of the full-length gene is about 7600. Under normal physiological conditions, the gene is transcribed and translated into proprocalcitonin in the rough endoplasmic reticulum next to the thyroid follicle. Calcitonin precursor contains 84 N-terminal amino acids, active calcitonin (CT, 32) peptide and ca...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577
CPCG01N33/54326G01N33/577
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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