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Chemiluminesent immunoassay kit for quantitatively detecting digoxin and preparation method of chemiluminesent immunoassay kit

A chemiluminescence immunoassay, detection kit technology, applied in chemiluminescence/bioluminescence, biological testing, analysis by chemical reaction of materials, etc., can solve the problem that the sensitivity and detection reliability cannot be guaranteed, and the detection system is expensive. Popularization and other issues, to achieve the effect of easy automation, unattended operation, and easy operation

Inactive Publication Date: 2019-12-27
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the price is cheap, the sensitivity and detection reliability cannot be guaranteed, and foreign detection systems are expensive and difficult to popularize

Method used

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  • Chemiluminesent immunoassay kit for quantitatively detecting digoxin and preparation method of chemiluminesent immunoassay kit
  • Chemiluminesent immunoassay kit for quantitatively detecting digoxin and preparation method of chemiluminesent immunoassay kit
  • Chemiluminesent immunoassay kit for quantitatively detecting digoxin and preparation method of chemiluminesent immunoassay kit

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preparation example Construction

[0041] The present invention also provides a preparation method of the above-mentioned chemiluminescent immunoassay kit for quantitatively detecting digoxin, comprising the following steps:

[0042] Step S1, preparation of reagent R1:

[0043] After mixing the streptavidin magnetic particle solution and TBST solution, place it on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, keep the magnetic particles, and prepare a solid phase reagent in the buffer after washing Wherein, the concentration of the streptavidin magnetic particle solution is 50-100 mg / ml; the volume ratio of the streptavidin magnetic particle solution and the TBST solution is preferably (0.5-1): (5-10) The mixing time is preferably 10-15min; the buffer is 50mM MES, 0.05% Tween-20, 0.05% Proclin300, pH6.5 or 100mM PBS, 0.1% Tween-20, 0.1% Proclin300 , pH7.2; the concentration of the solid-phase reagent is preferably 0.01% to 1%, more preferably 0.05%; the source of the...

Embodiment 1

[0051] Example 1 Preparation of chemiluminescent immunoassay kit for quantitative detection of digoxin

[0052] (1) Preparation of reagent R1

[0053] Take 0.5mL (50mg) of streptavidin magnetic particle solution with a concentration of 100mg / mL, the particle size of streptavidin magnetic particles is 0.05μm, add 10mL of TBST solution and mix well for 10min, place on a magnetic separator , until the supernatant has no turbidity, discard the supernatant, and keep the magnetic particles. After repeated washing for 3 times, use 50mM MES, 0.05% Tween, 0.05% Proclin300, pH 6.5 buffer to prepare a solid-phase reagent with a magnetic bead concentration of 0.05%, and store at 2-8°C.

[0054] (2) Preparation of reagent R2

[0055] Put 250μg digoxin monoclonal antibody into the centrifuge tube, make sure the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 20s), add PBS buffer solution, mix well, add 5mL 2mg / mL acridinium ester DMF after mixing The ...

Embodiment 2

[0058] Example 2 Preparation of Chemiluminescent Immunoassay Kit for Quantitative Detection of Digoxin

[0059] (1) Preparation of reagent R1

[0060] Take 0.72mL (72mg) of streptavidin magnetic particle solution with a concentration of 100mg / mL, the particle size of streptavidin magnetic particles is 3μm, add 15mL of TBST solution and mix well for 15min, place on a magnetic separator, Discard the supernatant until there is no turbidity in the supernatant, and keep the magnetic particles. After repeated washing 3 times, use 100mM PBS, 0.1% Tween-20, 0.1% Proclin300, pH 7.2 buffer solution to make a solid-phase reagent with a magnetic bead concentration of 0.072%, and store at 2-8°C.

[0061] (2) Preparation of reagent R2

[0062] Put 500μg digoxin monoclonal antibody into the centrifuge tube, make sure the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 30s), then add TRIS buffer solution, mix well, add 1mL 2.5mg / mL acridinium ester after...

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Abstract

The invention discloses a chemiluminescent immunoassay kit for quantitatively detecting digoxin. The chemiluminescent immunoassay kit is characterized in that the chemiluminescent immunoassay kit includes a reagent R1, a reagent R2 and a reagent R3, wherein the reagent R1 is a suspension containing streptavidin magnetic particles, the reagent R2 is a solution containing a digoxin monoclonal antibody marked by a chemiluminescent marker, and the reagent R3 is a solution containing digoxin derivatives marked by coupling markers. According to the chemiluminescent immunoassay kit provided by the invention, the digoxin is a small molecular substance, thus detection is conducted through the principle of a competitive method, the time and cost are saved, and the kit has the high specificity of animmune reaction, further has the high sensitivity of a luminous reaction and has the advantages of high sensitivity, high specificity, wide linear range, good reagent stability, easy and convenient operation and easy automation; a preparation method provided by the invention is simple in technology, and the kit is long in validity period and relatively low in cost; rapid detection is convenient torealize, the requirements for instruments are simple, and fully-automatic operation is convenient to realize.

Description

technical field [0001] The invention relates to the technical field of in vitro detection, in particular to a chemiluminescence immunoassay kit for quantitative detection of digoxin and a preparation method thereof. Background technique [0002] Digoxin (Digoxin, or DIG) is a medium-acting cardiac glycoside drug extracted from digitalis. It is a white crystal or crystalline powder, odorless, and bitter in taste. During treatment, the effect on the heart is manifested as positive inotropic effect, slowing down the heart rate, and inhibiting cardiac conduction. It is suitable for low-output congestive heart failure, atrial fibrillation, atrial flutter, and paroxysmal supraventricular tachycardia. speed. This positive inotropic effect makes digoxin an effective drug of choice in the treatment of heart failure to control rapid ventricular rate. [0003] However, the effective plasma concentration of digoxin is narrow, < 0.5 ng / ml in unmedicated patients. According to the 20...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/543G01N33/531G01N21/76
CPCG01N21/76G01N33/53G01N33/531G01N33/54326G01N33/9453
Inventor 李磊李冬梅韩美玉孙成艳高威何浩会
Owner DIRUI MEDICAL TECH CO LTD
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