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Neuron-specific enolase content detection kit and preparation method thereof

A detection kit, enolase technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of small application range, small measurement linear range, radioactive material pollution, etc., and achieve wide detection linear range, good bottle opening Stable, well-specific effects

Active Publication Date: 2015-08-26
CUSABIO TECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, clinical methods for the determination of neuron-specific enolase mainly include radioimmunoassay, ELLSA, and chemiluminescence. Among them, the degree of automation of the enzyme-linked immunoassay is not high, and it is greatly affected by human factors; the radioimmunoassay has radiation Contamination of substances; while the chemiluminescence method has high sensitivity, but the detection linear range is small and the detection cost is high, and a matching chemiluminescence detector is required. These reasons lead to a small range of applications
So far, there is a lack of a kit for detecting the content of neuron-specific enolase (NSE) with simple operation, high sensitivity and wide linear range in clinical practice.

Method used

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  • Neuron-specific enolase content detection kit and preparation method thereof
  • Neuron-specific enolase content detection kit and preparation method thereof
  • Neuron-specific enolase content detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Latex washing: Take 1mL of latex solution with a concentration of 10% and a particle size of 80nm (purchased from PolyMicrospheres) in a 50mL centrifuge tube, then add 9mL of PBS buffer solution with a pH of 7.0 to the centrifuge tube, shake and mix evenly, Then add 4 μL Tween-20 to the centrifuge tube, shake and mix evenly, place the centrifuge tube in a centrifuge at 25,000 rpm, centrifuge for 30 min, discard the supernatant, and redissolve with 9 mL of PBS buffer at pH 7.0. Sonicate for 60 seconds to disperse the latex particles evenly.

[0042] (2) Activated latex: Accurately weigh 50mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 50mg of N-hydroxysulfosuccinimide (NHS) and mix with 1mL0. Dissolve the 12M phosphate buffer solution, and add this solution to the latex solution in step (1), oscillate and mix evenly, and place on a constant temperature shaker at 25°C, 200 rpm, and react for 1h;

[0043] (3) Sensitized latex: Add 1mL of 1mg / mL neuron-s...

Embodiment 2

[0047] (1) Latex washing: Take 1 mL of latex solution with a concentration of 10% and a particle size of 240 nm (purchased from PolyMicrospheres) in a 50 mL centrifuge tube, then add 9 mL of PBS buffer solution with pH 7.0 to the centrifuge tube, shake and mix evenly, Then add 4 μL Tween-20 to the centrifuge tube, shake and mix evenly, place the centrifuge tube in a centrifuge at 25,000 rpm, centrifuge for 30 min, discard the supernatant, and redissolve with 9 mL of PBS buffer at pH 7.0. Sonicate for 60 seconds to disperse the latex particles evenly.

[0048] (2) Activated latex: Accurately weigh 50mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 50mg of N-hydroxysulfosuccinimide (NHS) and mix with 1mL0. Dissolve the 12M phosphate buffer solution, and add this solution to the latex solution in step (1), oscillate and mix evenly, and place on a constant temperature shaker at 25°C, 200 rpm, and react for 1h;

[0049] (3) Sensitized latex: Add 1mL1mg / mL neuron-speci...

Embodiment 3

[0053] (1) Latex washing: Take 1mL of latex solution with a concentration of 10% and a particle size of 150nm (purchased from Poly Microspheres) in a 50mL centrifuge tube, then add 9mL of PBS buffer solution with pH7.0 into the centrifuge tube, shake and mix well , and then add 4 μL Tween-20 to the centrifuge tube, shake and mix evenly, place the centrifuge tube in a centrifuge at 25,000 rpm, centrifuge for 30 min, discard the supernatant, and redissolve with 9 mL of PBS buffer at pH 7.0 , ultrasonic vibration for 60 seconds to disperse the latex particles evenly.

[0054] (2) Activated latex: Accurately weigh 50mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 50mg of N-hydroxysulfosuccinimide (NHS) and mix with 1mL0. Dissolve the 12M phosphate buffer solution, and add this solution to the latex solution in step (1), oscillate and mix evenly, and place on a constant temperature shaker at 25°C, 200 rpm, and react for 1h;

[0055] (3) Sensitized latex: Add 1mL of 1...

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Abstract

The invention discloses a neuron specific enolase content detection kit and a preparation method thereof, belonging to the field of human neuron specific enolase content detection. The detection kit is composed of mutually independent reagent I and reagent II, wherein the constituents of reagent I include a biological buffer, a preservative, a coagulation accelerator, inorganic salt and water; the constituents of the reagent II include latex particles of an envelope neuron specific enolase antibody, a biological buffer, a preservative, a suspending agent and water. According to the invention, the latex particles with particle size of 80-240nm are compositely enveloped by the neuron specific enolase monoclonal antibody and a polyclonal antibody, and thus, the kit is guaranteed to have high sensitivity and wide detection linear range, and has the advantages of being simple and convenient to operate, rapid, strong in specificity, excellent in stability and high in sensitivity, and so on.

Description

technical field [0001] The invention relates to a detection kit for enzyme content in human body, in particular to a detection kit for neuron-specific enolase content in human body and a preparation method thereof, belonging to the detection field of human neuron-specific enolase content. Background technique [0002] Small cell lung cancer (Small Cell Lung Cancer, SCLC) accounts for about 20% of lung cancers. It has a high degree of malignancy, a short doubling time, early and extensive metastasis, is sensitive to chemotherapy and radiotherapy, and has a high remission rate after initial treatment, but it is very prone to secondary disease. Drug resistance, easy to relapse, the main treatment is systemic chemotherapy. If high-risk patients can be identified early after onset and treated in time, the mortality and prognosis of small cell lung cancer will be significantly improved. Therefore, tumor markers play an important role in the diagnosis and prognosis of SCLC. [00...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573G01N33/531
Inventor 华权高沈鹤霄许可闫彩霞黄爱伍卫姣舒芹鄢宝赵畅
Owner CUSABIO TECH LLC
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