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Kit for quantitatively detecting procalcitonin and preparation method thereof

A technology for quantitative detection of procalcitonin, applied in the field of biomedicine, can solve the problems of low detection sensitivity, narrow inspection range, poor specificity, etc., and achieve the effects of high sensitivity, high sensitivity and high accuracy

Inactive Publication Date: 2018-06-15
浙江艾明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

[0004] By providing a kit and preparation method for the quantitative detection of procalcitonin, this solution solves the shortcomings of low detection sensitivity, poor specificity, and narrow detection range in the prior art

Method used

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  • Kit for quantitatively detecting procalcitonin and preparation method thereof
  • Kit for quantitatively detecting procalcitonin and preparation method thereof
  • Kit for quantitatively detecting procalcitonin and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preparation of PCT alkaline phosphatase-labeled monoclonal antibody

[0050] 1) Add 100 μL alkaline phosphatase to 300 μL 0.01M PBS with pH7.4;

[0051] 2) Add 40 μL of glutaraldehyde with a volume ratio of 25%, mix well, and activate at 2-8°C for 16 hours;

[0052] 3) Transfer the reaction solution to a dialysis bag (molecular weight cut-off 8000-12000), dialyze in 0.01M PBS with pH7.4 for 24 hours, and change the solution twice in the middle;

[0053] 4) Take an appropriate amount of monoclonal antibody to procalcitonin, and make a 4mg / mL solution with 0.01M PBS at pH7.4;

[0054] 5) Take 125 μL of the solution in step 4 and add it to the dialysis bag in step 3, and react at 2-8°C for 24 hours;

[0055] 6) Transfer the solution to a glass bottle, add an equal volume of glycerol, and store at -15°C for later use.

Embodiment 2

[0056] Example 2 Preparation of the PCT quantitative assay kit of the present invention

[0057] 1. Preparation of alkaline phosphatase antibody

[0058] 1. Alkaline phosphatase labeled monoclonal antibody diluent:

[0059] Casein (Casein) 0.2% (weight)

[0060] Tris-HCl buffer 0.02M, pH7.4.

[0061] 2. Select the concentration of alkaline phosphatase-labeled monoclonal antibody:

[0062] The working concentration of alkaline phosphatase-labeled monoclonal antibody is greater than 1.0 μg / mL.

[0063] 2. Preparation of PCT calibrator

[0064] Prepared with pure PCT, diluted with PBS buffer containing 2% bovine serum albumin (BSA) by weight, 0.02M pH7.4, divided into 0.5mL / bottle, the concentration is 0, 0.2, 0.5, 2, 5 , 20ng / mL, a total of 6 vials, stored at 2-8°C.

[0065] 3. Preparation of alkaline phosphatase-labeled antibody:

[0066] (1) Add 100 μL of alkaline phosphatase to 300 μL of 0.01M PBS at pH 7.4;

[0067] (2) Add 40 μL of glutaraldehyde with a volume ratio...

Embodiment 3

[0097] Embodiment 3 Proportion of chemiluminescent substrate solution

[0098] Chemiluminescence refers to the emission of light accompanying the chemical reaction process. During the chemical reaction, certain substances (luminescent agents) absorb the chemical energy generated during the reaction, causing the electrons in the product molecules of the reaction or the intermediate state molecules of the reaction to jump to the excited state. When the electrons return from the excited state to the ground state , energy is released in the form of photons.

[0099] The ratio of chemiluminescent substrate solution has a great influence on the sensitivity of PCT quantitative detection.

[0100] Wherein, on the basis of the above-mentioned technical scheme, preferably described chemiluminescence substrate liquid contains:

[0101] AMPPD 20% (volume)

[0102] NaCl 16% (weight)

[0103] KCl 0.5% (weight)

[0104] Tris-HCl buffer 0.2M pH 8.0~10.0;

[0105] Among them, the differe...

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Abstract

The invention provides a kit for quantitatively detecting procalcitonin and a preparation method thereof. The kit comprises an alkaline phosphatase labeled monoclonal antibody of procalcitonin, a biotin labeled monoclonal antibody of procalcitonin, a carrier with coated streptavidin, a procalcitonin calibration product, and a chemiluminescent substrate for alkaline phosphatase. At the same time, the kit preparation method comprises following steps: labeling a procalcitonin monoclonal antibody by alkaline phosphatase; labeling a procalcitonin monoclonal antibody by biotin; coating a carrier bya procalcitonin monoclonal antibody; preparing a procalcitonin calibration product by using pure procalcitonin; split-charging the calibration product, the alkaline phosphatase labeled procalcitonin monoclonal antibody, the biotinylated antibody, and a chemiluminescent substrate; and carrying out assembly to obtain a finished product. The provided kit has the advantages of high sensitivity, good specificity, high accuracy of detection results of quantitative detection, low using cost, and easiness for popularization and application.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a kit for quantitatively detecting procalcitonin and a preparation method thereof. Background technique [0002] Procalcitonin (PCT, procalcitonin) is a protein. In normal and healthy individuals, its serum concentration is extremely low, only 10-50pg / ml, which cannot be detected by general methods. In patients with systemic bacterial, fungal and parasitic infections, systemic inflammatory response syndrome, sepsis, acute and chronic pneumonia, acute pancreatitis, active hepatitis, trauma, etc., the serum PCT level is abnormally increased, and its concentration is several times or even higher than the normal level of HKUST. Tens of thousands of times; in patients with viral infections, autoimmune diseases, and organ transplant rejection reactions, the serum concentration does not increase or slightly increases. This determines the high specificity of PCT, so it can be used in the diff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/535
CPCG01N33/535G01N33/577
Inventor 胡国富朱炎
Owner 浙江艾明德生物科技有限公司
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