Reagent for determining procalcitonin and preparation method of reagent

A procalcitonin, human calcitonin technology, applied in biological testing, material analysis by observing the effect on chemical indicators, measuring devices, etc. Burden and other problems, to achieve the effect of improving detection efficiency, rapid detection, and high accuracy

Active Publication Date: 2013-04-03
YESEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are two kinds of kits in the market, tube type and plate type, but this method has the disadvantages of low luminescence efficiency and unstable labeling of acridinium ester, luminol, and isoluminol directly labeled antibodies; and this direct labeling method Belongs to the instant light-emitting type, it is difficult to guarantee the stability and repeatability of the test results, and special testing instruments are required
Not convenient for routine laboratory development
However, the electrochemiluminescence immunoassay kit for detecting PCT using the biotin-avidin system needs to be equipped with an expensive automatic electrochemiluminescence detector, which cannot be carried out in conventional laboratories, and it will bring unnecessary costs to patients and increase the burden on patients.
ELISA uses artificially synthesized PCT monoc

Method used

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  • Reagent for determining procalcitonin and preparation method of reagent
  • Reagent for determining procalcitonin and preparation method of reagent
  • Reagent for determining procalcitonin and preparation method of reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] 1) Procalcitonin Reagent 1

[0057]

[0058] After dissolving the above raw materials with 800ml of purified water, add purified water to make the volume to 1000ml, and then adjust the pH to 7.2 with hydrochloric acid;

[0059] 2) Procalcitonin Reagent 2

[0060] 1. Take 80nm carboxyl latex microspheres and dilute them with 50mmol / L, pH6.0 MES buffer to form a suspension of 0.01mg / ml;

[0061] 2. Add 20 mg of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) and 40 mg of sodium N-hydroxysuccinimide sulfonate to 1 ml of carboxyl latex microsphere suspension Add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide sulfonate sodium salt, mix immediately and place at room temperature The mixture was reacted for 15-30 minutes, stirring constantly;

[0062] 3. Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purifie...

Embodiment 2

[0077] 1) Procalcitonin Reagent 1

[0078]

[0079] After dissolving the above raw materials with 800ml of purified water, add purified water to make the volume to 1000ml, and then adjust the pH to 8.0 with sodium hydroxide;

[0080] 2) Procalcitonin Reagent 2

[0081] 1. Dilute 150nm carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer to a 0.01mg / ml suspension;

[0082] 2. Add EDAC and NHS according to the ratio of 1ml carboxyl latex microsphere suspension plus 20mg EDAC and 40mg NHS, mix immediately and react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0083] 3. Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water, remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a suspension of carboxyl latex microspheres, wherein the carboxyl latex microspheres The content of latex microspheres is 0.01mg / ml;

[0084] 4. Dissolve the rabbit anti-human procalcitonin anti...

Embodiment 3

[0098] 1) Procalcitonin Reagent 1

[0099]

[0100] After dissolving the above raw materials with 800ml of purified water, add purified water to make the volume to 1000ml, and then adjust the pH to 8.0 with sodium hydroxide;

[0101] 2) Procalcitonin Reagent 2

[0102] 1. Take 300nm carboxyl latex microspheres and dilute them with 50mmol / L, PH6.0 MES buffer to form a suspension of 0.01mg / ml;

[0103] 2. Add EDAC and NHS according to the ratio of 1ml carboxyl latex microsphere suspension plus 20mg EDAC and 40mg NHS, mix immediately and react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0104] 3. Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a suspension of carboxyl latex microspheres, in which carboxyl The content of latex microspheres is 0.01mg / ml;

[0105] 4. Dissolve the mouse anti-human procalcitonin ant...

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Abstract

The invention relates to a reagent for determining procalcitonin and a preparation method of the reagent, in order to provide the reagent which has the characteristic of high accuracy degree, and the provided preparation method has the characteristic of convenience in preparation. The technical scheme is that the reagent for determining procalcitonin comprises the following components: a. a procalcitonin reagent 1, b. a procalcitonin reagent 2, and c. a liquid-type procalcitonin reference calibration item. The preparation method of the reagent for determining procalcitonin comprises the following steps of: (1) uniformly mixing the procalcitonin reagent 1; (2) removing the procalcitonin reagent 2, the removing process specifically comprises the following steps of: a. taking a suspension, b. carrying out reaction on a mixture, c. acquiring a carboxylic latex microsphere suspension, d. regulating the concentration, e. taking the suspension in step (3), and adding into the step (4), f. reacting, g. adding cholamine, and h. carrying out centrifugal treatment; and (3) treating the liquid-type procalcitonin reference calibration product: mixing according to the amount of a formula, and ranking according to the content of the procalcitonin or adding a procalcitonin pure product in the mixed liquid.

Description

technical field [0001] The invention relates to a reagent for measuring procalcitonin and a preparation method of the reagent, in particular to a reagent for detecting procalcitonin in serum or plasma by latex-enhanced immune turbidimetry and a preparation method, which can be widely used in medicine and biochemical technology. Background technique [0002] Serum procalcitonin (Procalcitionin, PCT) is composed of 116 amino acids, a glycoprotein with a molecular weight of 13KD. Its half-life is 25-30 hours. The plasma PCT content of healthy people is extremely low, lower than 0.5ng / ml in normal human blood. Its level in plasma increases when severe bacterial, fungal, and parasitic infections, as well as sepsis and multiple organ failure; autoimmunity, allergies, and local infections, viral infections, chronic non-specific inflammation, cancer fever PCT levels will not increase in diseases such as graft host rejection and so on; this determines the high specificity of PCT, ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531G01N21/82
Inventor 陈开华其他发明人请求不公开姓名
Owner YESEN BIOTECH SHANGHAI
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