cTnI detection kit and using method thereof

A detection kit and magnetic bead technology, which is applied in the field of immunochemical detection, can solve the problems that cannot be realized, achieve the effects of improving detection speed, simplifying operation steps, and realizing high sensitivity and wide range detection

Active Publication Date: 2018-04-17
NANTONG EGENS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, the technical problem to be solved by the present invention is to overcome the defect that cTnI detection in the prior art cannot realize the detection of whole blood samples

Method used

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  • cTnI detection kit and using method thereof

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Embodiment 1

[0056] This embodiment provides a method for preparing an alkaline phosphatase-labeled cTnI antibody, comprising the following steps:

[0057] 1. The storage concentration of cTnI-labeled antibody (i.e. cTnI antibody) is 2mg / mL, add 0.5M TECP solution to the cTnI-labeled antibody until the final concentration of TECP-labeled antibody is 1.25mM, mix immediately, and stand at 25°C for reaction 60 minutes.

[0058] 2. The cTnI-labeled antibody solution obtained in step 1 was immediately desalted, replaced with a pH 7.3, 100 mM triethanolamine solution, and the concentration of the antibody solution was measured by ultraviolet spectrophotometry.

[0059] 3. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.

[0060] 4. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml), the molar ratio of alkaline phosphatase to Sulfo...

Embodiment 2

[0069] This embodiment provides a method for preparing an alkaline phosphatase-labeled cTnI antibody, comprising the following steps:

[0070] 1. The storage concentration of cTnI-labeled antibody (i.e. cTnI antibody) is 2mg / mL, add 0.5M TECP solution to the cTnI-labeled antibody until the final concentration of TECP-labeled antibody is 1.25mM, mix immediately, and stand at 25°C for reaction 60 minutes.

[0071] 2. The cTnI-labeled antibody solution obtained in step 1 was immediately desalted, replaced with a pH 7.3, 100 mM triethanolamine solution, and the concentration of the antibody solution was measured by ultraviolet spectrophotometry.

[0072] 3. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.

[0073] 4. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml), the molar ratio of alkaline phosphatase to Sulfo...

Embodiment 3

[0082] This embodiment provides a method for preparing cTnI antibody-labeled magnetic beads, comprising the following steps:

[0083] 1. Store the cTnI-coated antibody (i.e., cTnI antibody) in MES buffer at pH 5.0 and 100 mM, and the antibody concentration is 1 mg / mL;

[0084] 2. Take the carboxyl magnetic bead solution 100 times the weight of the antibody, the concentration of the carboxyl magnetic bead solution is 100 mg / ml, the particle size of the carboxyl magnetic bead is 3 μm, and the supernatant is discarded by magnetic separation;

[0085] 3. Washing: re-dissolve the magnetic beads obtained in the above steps with pH 5.0, 100 mM MES buffer, and discard the supernatant by magnetic separation.

[0086] 4. Repeat step 3 once.

[0087] 5. Add an appropriate amount of 100mM MES buffer to dissolve the magnetic beads obtained in step 4.

[0088] 6. Weigh an appropriate amount of NHS (N-hydroxysuccinimide) reagent and dissolve it with pH 5.0, 100mM MES buffer to form a solut...

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Abstract

The invention discloses a cTnI detection kit which comprises a calibration product, a cleanout fluid, a substrate solution, a pretreatment liquid, an enzyme conjugate working solution and a magnetic bead conjugate working solution, wherein the pretreatment liquid contains imidazole; the enzyme conjugate working solution contain an enzyme labeled cTnI antibody; and the magnetic bead conjugate working solution contains cTnI antibody labeled magnetic beads. The cTnI detection kit can realize accurate determination of cTnI in a whole blood sample, the detection steps are simplified, and the detection efficiency is improved. The lowest detection limit of the kit is 0.02ng / ml, the linear range is 0.02-50ng / ml, the detection sensitivity is high, the linear range is wide, and the detection resultis accurate. The invention further discloses a using method of the cTnI detection kit. The using method is simple in using steps, the detection time of the cTnI is effectively shortened, and rapid andsensitive detection of the cTnI is realized.

Description

technical field [0001] The invention belongs to the technical field of immunochemical detection, and in particular relates to a cTnI detection kit and a use method thereof. Background technique [0002] Cardiovascular disease is a major fatal disease that endangers the health and life of the people in our country. According to the statistics of the Ministry of Health, in my country's cities, 235 people die of cardiovascular and cerebrovascular diseases for every 100,000 people, accounting for 100% of all diseases. The number of deaths caused by the disease is the first, and it is increasing at a rate of 2% every year. Acute Myocardial Infarction (AMI) is the main cause of death of cardiovascular patients. AMI is a clinically sudden disease. Its onset is often accompanied by a series of symptoms such as chest pain, nausea, fever, arrhythmia, and elevated serum markers. Early diagnosis and interventional treatment will help save dying myocardium and improve prognosis. It is o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/535
CPCG01N33/535G01N33/54326G01N33/6887G01N2333/4712G01N2446/90G01N2800/324
Inventor 王保君汤双双欧卫军徐艳
Owner NANTONG EGENS BIOTECH CO LTD
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