Myoglobin detection reagent kit and method for applying same

A technology for detecting kits and myoglobin, applied in the field of myoglobin detection kits, can solve the problems of narrow detection range, low specificity of myoglobin detection kits, cumbersome operation, etc., and achieve detection sensitivity and precision Improve the effect of avoiding non-specific binding and simplifying the operation steps

Active Publication Date: 2018-04-13
NANTONG EGENS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Therefore, the technical problem to be solved by the present invention is that the existing myoglobin detection kit has low specificity, and adopts plasma or serum as a sample, has the defects of cumbersome operation and narrow detection range, thereby providing a high specificity, capable A myoglobin detection kit that directly uses whole blood as a sample; further, the present invention also provides a method for using the detection kit

Method used

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  • Myoglobin detection reagent kit and method for applying same

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Embodiment 1

[0055] The preparation of embodiment 1 enzyme conjugate

[0056] 1. Weigh 3mg of Traut's reagent and prepare a solution with a concentration of 1.376mg / mL with 100mmol / L triethanolamine buffer solution (pH=8.5±0.05); L of triethanolamine buffer solution (pH = 8.5 ± 0.05) was prepared into a solution with a concentration of 2 mg / mL, and was added with a concentration of 1.376 mg / mL of Traut's solution, the mixture of myoglobin-labeled antibody and Traut's reagent The molar ratio is 1:15, mix immediately, and react at room temperature for 15 minutes. Add the glycine solution of 1mmol / L, the amount of substance of glycine is 10 of the amount of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide ester sodium salt substance times, mix immediately, and react at room temperature for 10 minutes. Desalted and replaced with 100 mmol / L triethanolamine cross-linking buffer (pH=7.3±0.05).

[0057] 2. Weigh 3 mg of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sul...

Embodiment 2

[0061] The preparation of embodiment 2 enzyme conjugates

[0062] 1. Weigh 3mg of Traut's reagent and prepare a solution with a concentration of 1.3mg / mL with 100mmol / L triethanolamine buffer solution (pH=8.5±0.05); L of triethanolamine buffer solution (pH=8.5 ± 0.05) was prepared into a solution with a concentration of 5 mg / mL, and was added with a concentration of 1.3 mg / mL Traut's solution, the mixture of myoglobin-labeled antibody and Traut's reagent The molar ratio is 1:10, mix immediately, and react at room temperature for 12 minutes. Add the glycine solution of 1mmol / L, the amount of substance of glycine is 20% of the amount of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide ester sodium salt substance times, mix immediately, and react at room temperature for 12 minutes. Desalted and replaced with 100 mmol / L triethanolamine cross-linking buffer (pH=7.3±0.05).

[0063] 2. Weigh 3 mg of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosucci...

Embodiment 3

[0067] The preparation of embodiment 3 enzyme conjugates

[0068] 1. Weigh 3mg of Traut's reagent and prepare a solution with a concentration of 1.5mg / mL with 100mmol / L triethanolamine buffer solution (pH=8.5±0.05); L of triethanolamine buffer solution (pH = 8.5 ± 0.05) was prepared into a solution with a concentration of 2.5 mg / mL, and to it was added a concentration of 1.5 mg / mL Traut's solution, myoglobin-labeled antibody and Traut's reagent The molar ratio is 1:15, mix immediately, and react at room temperature for 18 minutes. Add the glycine solution of 1mmol / L, the amount of substance of glycine is 15% of the amount of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide ester sodium salt substance times, mix immediately, and react at room temperature for 10 minutes. Desalted and replaced with 100 mmol / L triethanolamine cross-linking buffer (pH=7.3±0.05).

[0069] 2. Weigh 3 mg of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide sodiu...

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Abstract

The invention belongs to the field of in-vitro diagnostic reagent kits, and particularly relates to a myoglobin detection reagent kit and a method for applying the same. The myoglobin detection reagent kit comprises calibration products, reagents R1, enzyme conjugate working solution R2, magnetic bead conjugate working solution M, cleaning solution and chemiluminescent substrates. The reagents R1contain imidazole components, blood cells in whole blood can be quickly removed by the imidazole components, and accordingly the possibility that magnetic beads are swallowed by the blood cells can beeffectively prevented; the cleaning solution contains sodium lauryl sulfate components, accordingly, cleaning effects can be enhanced, and non-specific binding of the chemiluminescent substrates canbe prevented. The myoglobin detection reagent kit and the method have the advantages that finger peripheral whole blood or anticoagulant venous whole blood can be directly used as a to-be-detected sample for the myoglobin detection reagent kit, the whole blood sample can be directly detected without being pretreated, accordingly, the detection speed can be greatly increased, operation steps can besimplified, the application range of the myoglobin detection reagent kit can be expanded, and large-scale popularization and application can be facilitated.

Description

technical field [0001] The invention belongs to the field of in vitro diagnostic kits, and in particular relates to a myoglobin detection kit and a use method thereof. Background technique [0002] Myoglobin (MB) is a specific oxygen-conjugate protein for skeletal muscle and cardiac muscle. It consists of a polypeptide chain with 153 amino acid residues and a blood-red prosthetic group. The molecular weight is 17.8kD. function of transporting and storing oxygen. Normal human cardiac muscle and skeletal muscle contain a large amount of MB but very little in the blood, but when the cardiac muscle or skeletal muscle is damaged, MB will be released from the muscle tissue into the circulating blood, filtered by the glomerulus, and appear in the urine. Therefore, the determination of MB in blood and urine can be used for the diagnosis of some myopathy and heart disease, such as acute myocardial infarction (AMI), acute muscle injury, acute and chronic renal failure, severe congest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/535G01N21/76
CPCG01N21/763G01N33/535G01N33/54326G01N33/6887G01N2333/4712G01N2333/805G01N2446/90
Inventor 王保君汤双双欧卫军褚晖唐启伟顾一峰
Owner NANTONG EGENS BIOTECH
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