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Trypsinogen-2 detection kit and preparation method thereof

A trypsinogen and detection kit technology, which is applied to measurement devices, instruments, scientific instruments, etc., can solve problems such as long time consumption, cumbersome detection process, and inability to quantitatively detect, and achieves guaranteed stability, guaranteed sensitivity, and good specificity. Effect

Active Publication Date: 2013-09-18
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA has high accuracy and good sensitivity in detecting samples, but the detection process is cumbersome and time-consuming, and samples need to be tested in batches, which is not suitable for timely inspection; meanwhile, ELISA detection has a low degree of automation, and the detection results are affected Human factors have a greater influence
Immunochromatography is a rapid diagnostic technique that has emerged abroad in recent years. This method detects samples quickly, but generally cannot be used for quantitative detection and can only be used for qualitative judgment.

Method used

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  • Trypsinogen-2 detection kit and preparation method thereof
  • Trypsinogen-2 detection kit and preparation method thereof
  • Trypsinogen-2 detection kit and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Preparation of latex particles coated with trypsinogen-2 antibody in preliminary example 1

[0039] (1) Preparation of latex solution: take 1g of polystyrene latex microsphere particles with a particle size of 200nm, and disperse the latex particles with 100mL of 0.05mol / L 2-(N-morpholine)ethanesulfonic acid buffer solution, latex microsphere particles The final concentration is 1% (W / V);

[0040] (2) Add 10 mg of N-hydroxysuccinimide to the latex solution in step (1) to a final concentration of 0.1 mg / mL; add 50 mg of 1-ethyl-3-(3-dimethylaminopropyl ) carbodiimide, the final concentration is 0.5mg / mL, stirring to fully dissolve each substance, stirring the mixture at room temperature for 30 minutes, after the reaction is completed, centrifuge the reaction system at 18000 rpm for 30min, discard supernatant;

[0041] (3) Wash the precipitate in step (2) with 0.05mol / L 2-(N-morpholine)ethanesulfonic acid buffer to remove unreacted N-hydroxysuccinimide and 1-ethyl-3-(3-...

Embodiment 2

[0045] Preparation of latex particles coated with trypsinogen-2 antibody in preliminary example 2

[0046] (1) Preparation of latex solution: Take 0.8g of polystyrene latex microsphere particles with a particle size of 150nm, and disperse the latex particles with 100mL of 0.05mol / L 2-(N-morpholine)ethanesulfonic acid buffer solution, latex microspheres The final concentration of particles is 0.8% (W / V);

[0047] (2) Add 15 mg of N-hydroxysuccinimide to the latex solution in step (1) to a final concentration of 0.15 mg / mL; add 40 mg of 1-ethyl-3-(3-dimethylaminopropyl ) carbodiimide, the final concentration is 0.4mg / mL, stirring to fully dissolve each substance, stirring the mixture at room temperature for 30 minutes, after the reaction is completed, centrifuge the reaction system at 20000 rpm for 20min, discard supernatant;

[0048] (3) Wash the precipitate in step (2) with 0.05mol / L 2-(N-morpholine)ethanesulfonic acid buffer to remove unreacted N-hydroxysuccinimide and 1-et...

Embodiment 3

[0052] Preliminary Example 3 Preparation of latex particles coated with trypsinogen-2 antibody

[0053] (1) Preparation of latex solution: Take 1.2g of polystyrene latex microsphere particles with a particle size of 250nm, and disperse the latex particles with 100mL of 0.05mol / L 2-(N-morpholine)ethanesulfonic acid buffer solution, latex microspheres The final concentration of particles is 1.2% (W / V);

[0054] (2) Add 20 mg of N-hydroxysuccinimide to the latex solution in step (1) to a final concentration of 0.2 mg / mL; add 50 mg of 1-ethyl-3-(3-dimethylaminopropyl ) carbodiimide, the final concentration is 0.5mg / mL, stirring to fully dissolve each substance, stirring the mixture at room temperature for 40 minutes, after the reaction is completed, centrifuge the reaction system at 18000 rpm for 30min, discard supernatant;

[0055] (3) Wash the precipitate in step (2) with 0.05mol / L 2-(N-morpholine)ethanesulfonic acid buffer to remove unreacted N-hydroxysuccinimide and 1-ethyl-...

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Abstract

The invention discloses a trypsinogen-2 content detection kit and a preparation method thereof. The detection kit consists of reagent I and reagent II which are independent of each other, wherein the reagent I comprises the following components: a biological buffer, a preservative, inorganic salt and water; the reagent II comprises the following components: latex granules coated by trypsinogen-2 antibody, the biological buffer, the preservative and water. The trypsinogen-2 content detection kit adopts the latex granules with the particle size of 100-300nm to prepare the latex granules coated by the trypsinogen-2 antibody, thus effectively guaranteeing the sensitivity of the kit, the detection linearity range, the measurement value accuracy and the bottle opening stability; furthermore, the latex granules coated by the antibody are prepared by a chemical cross-linking method, so that the combination stability between the antibody and the latex granules can be guaranteed, and the stability of the detection kit is improved. The detection kit has the advantages of being good in stability, high in sensitivity, high in specificity, simple and convenient to operate, etc.

Description

technical field [0001] The present invention relates to a test kit for detecting trypsinogen-2 content, in particular to a test kit for detecting trypsinogen-2 content in the human body and a preparation method thereof. The method for using the content of 2 belongs to the detection field of trypsinogen-2 content in human body. Background technique [0002] Acute pancreatitis (AP) is one of the common acute abdomens in clinical practice. According to foreign statistics, its incidence is 6.5-80 / 100,000 people, accounting for about 10% of acute abdomens. Some scholars believe that its pathogenesis is related to the excessive activation of trypsin caused by genetic changes. The onset of this disease is acute and the condition is dangerous. About 20% to 30% of AP cases can be transformed into necrotizing pancreatitis, leading to severe complications such as sepsis. Therefore, early diagnosis and treatment are very important. Blood amylase (AMS) is a commonly used detection item...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/531
Inventor 华权高许可沈鹤霄黄爱来祥兵伍卫姣舒芹
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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