CN-122017222-A - Application of improved isoluminol luminous substrate in MaglumiX chemiluminescent immunoassay system

Abstract

The invention discloses an application of an improved isoluminol luminous substrate in MaglumiX chemiluminescent immunoassay systems, and belongs to the technical field of in-vitro diagnostic reagents. The luminescent substrate comprises an alkaline solution containing a metal leaf-ine complex and derivatives thereof, piperazine phenol derivatives and a peroxide solution containing urea and stannate. The invention adds metal leaf-shaped substances into alkaline solution, stabilizes an active structure through coordination modification, promotes the reaction with hydrogen peroxide, enhances a chemiluminescence signal, improves the luminous efficiency and the detection sensitivity, and adds piperazine phenol derivatives to realize the persistent glow of isoluminol, ensure the signal stability and improve the result reliability. In the hydrogen peroxide system, urea/stannate stabilizer is added to raise the stability of the substrate. Experiments prove that the substrate has high accuracy correlation with the original reagent, excellent precision and stable performance after being stored for 24 months at room temperature, and is suitable for the detection fields of hormone, tumor markers and the like.

Inventors

• XIE QINGQING
• HU LIMIN

Assignees

• 浙江鑫科医疗科技有限公司

Dates

Publication Date

20260512

Application Date

20260211

Claims (10)

1. 1. The application of the improved isoluminol luminous substrate in MaglumiX-8 chemiluminescent immunoassay system is characterized in that the luminous substrate comprises a first substrate liquid and a second substrate liquid, wherein the first substrate liquid comprises alkali solution, a luminous enhancer 1-20mg/L, a luminous stabilizer 1-20mmol/L, a surfactant 0.1-5g/L, a buffering agent 0.01-0.10mol/L, a pH value is 13.0, the second substrate liquid comprises peroxide 0.5-2.0g/L, a buffering agent 0.01-0.10mol/L, a peroxide stabilizer 0.5-5.0g/L and a pH value is 5.30.
2. 2. The use of the improved isoluminol luminescent substrate according to claim 1 in MaglumiX chemiluminescent immunoassay systems wherein the alkaline solution in substrate solution one is 0.4mol/L aqueous sodium hydroxide solution.
3. 3. The use of an improved isoluminol luminescence substrate according to claim 1 in MaglumiX chemiluminescent immunoassay system, wherein the luminescence enhancer is one of chlorinated metal leaf-in, histidine/tryptophan/phenylalanine modified metal leaf-in/chlorinated metal leaf-in.
4. 4. The method for preparing the improved isoluminol luminous substrate in MaglumiX chemiluminescent immunoassay systems according to claim 1, wherein the luminous stabilizer is one of piperazine phenol derivatives such as N-methylpiperazine phenol, N-acetylpiperazine phenol, piperazine phenol hydrochloride and the like in substrate solution I.
5. 5. The use of the improved isoluminol luminous substrate according to claim 1 in MaglumiX chemiluminescent immunoassay system, wherein the surfactant is one of sodium dodecyl sulfate, tween-20, tween-80, triton X-100 and Brij-35.
6. 6. The use of the improved isoluminol luminous substrate according to claim 1 in MaglumiX chemiluminescent immunoassay systems, wherein the buffer is one or more of carbonate buffer, borate buffer and glycine in substrate solution one.
7. 7. The use of the improved isoluminol luminous substrate according to claim 1 in MaglumiX chemiluminescent immunoassay systems, wherein the peroxide in the substrate solution II is one of carbamide peroxide, hydrogen peroxide and sodium perborate.
8. 8. The use of the improved isoluminol luminous substrate in MaglumiX chemiluminescent immunoassay system according to claim 1, wherein the buffer is one or more of acetic acid, sodium acetate, citric acid, sodium citrate, sodium dihydrogen phosphate and disodium hydrogen phosphate.
9. 9. The use of the improved isoluminol luminous substrate according to claim 1 in MaglumiX chemiluminescent immunoassay systems, wherein the peroxide stabilizer in the substrate solution II is one or more of uric acid, stannate and organic phosphonate.
10. 10. The application of the improved isoluminol luminous substrate in MaglumiX chemiluminescent immunoassay systems according to claim 1 is characterized in that one component of the substrate solution is 0.4mol/L of sodium hydroxide, 5mg/L of histidine-coordinated ferric chloride leaf, 10mmol/L of N-acetylpiperazine phenol, 201g/L of Tween-and 0.05mol/L of sodium carbonate-sodium bicarbonate buffer solution, and two components of the substrate solution are 0.5g/L of hydrogen peroxide, 0.05mol/L of acetic acid-sodium acetate buffer solution and 2.0g/L of urea.

Description

Application of improved isoluminol luminous substrate in MaglumiX chemiluminescent immunoassay system Technical Field The invention belongs to the technical field of in-vitro diagnostic reagents, and particularly relates to an application of an improved isoluminol luminescent substrate in MaglumiX chemiluminescent immunoassay systems. Background The chemiluminescence immunoassay (ChemiluminescenceImmunoassay, CLIA) is a detection method combining a chemiluminescence technology and an immunoassay principle, has the advantages of high sensitivity, strong specificity, low background signal, high detection speed, simple operation, high automation degree and the like, and is widely applied to the fields of clinical diagnosis, environmental monitoring, food safety and the like. According to market data, the in-vitro diagnosis market scale of China exceeds 700 hundred million yuan, the annual compound growth rate reaches more than 18%, the CLIA is taken as the fastest growing sub-industry, and the imported substitution space is huge. Common chemiluminescent labels include acridinium esters and derivatives thereof, luminol (luminol) and derivatives thereof (e.g., isoluminol), adamantyl dioxetane, and the like. These labels may bind to enzymes (e.g., horseradish peroxidase HRP or alkaline phosphatase ALP) or act directly as luminescent substrates and undergo a chemical reaction under the action of an oxidizing agent (e.g., hydrogen peroxide) to produce a light signal that can be captured by a photomultiplier tube. The isoluminol (isoluminol) luminescent system is used as a core component in CLIA, and is usually combined with HRP enzyme through isoluminol and derivatives thereof to generate glow-type chemiluminescence in the presence of an oxidant, so that quantitative detection of analytes is realized. The system has high signal-to-noise ratio and wide linear range (which can exceed the upper limit of ELISA (enzyme-linked immunosorbent assay)) and can detect trace analytes with high sensitivity, complete signal generation and reading in a short time, and be beneficial to rapid diagnosis. The isoluminol luminous system is widely applied to the fields of hormone detection, tumor markers, infectious diseases, autoimmune diseases, myocardial markers and the like. The novel industry MaglumiX full-automatic chemiluminescence immunoassay instrument is used as high-performance CLIA equipment, supports the flux of 600 tests/hour, has the initial result time of only 15 minutes, stands by for 24 hours, supports 2800 continuous tests, and is suitable for medium and large laboratories. However, the prior isoluminol luminous system has the limitations that part of the system has shorter signal duration (glow type reaction is easy to attenuate) and can influence the stability of signal reading, and the equipment is required to have quick response capability; in addition, chemiluminescent reagents such as isoluminol conjugates and hydrogen peroxide are active in chemical property, sensitive to environmental factors such as illumination, temperature and the like, poor in storage stability (the shelf life is usually 6-18 months), easy to generate free radical-induced enzyme inactivation or substrate decomposition, and limit the clinical application efficiency and reliability of the chemiluminescent reagents. Disclosure of Invention 1. Problems to be solved Aiming at the problems existing in the prior art, the invention provides an application of an improved isoluminol luminous substrate in a MaglumiX chemiluminescent immunoassay system, and the substrate realizes luminous signal enhancement, continuous glow and substrate stability improvement by optimizing a metal leaf-ine complex and piperazine phenol derivative in an alkaline solution and a urea/stannate stabilizer in a peroxide solution, so that the detection sensitivity, the result reliability and the reagent shelf life are improved, and the substrate is suitable for the field of external diagnosis such as clinical hormone, tumor markers and the like. 2. Technical proposal In order to solve the problems, the invention adopts the following technical scheme. The substrate solution comprises a first substrate solution and a second substrate solution, wherein the first substrate solution comprises 1-20mg/L of an alkali solution, 1-20mmol/L of a luminescence enhancer, 0.1-5g/L of a luminescence stabilizer, 0.01-0.10mol/L of a surfactant and 13.0 of pH value, the second substrate solution comprises 0.5-2.0g/L of peroxide, 0.01-0.10mol/L of a buffer, 0.5-5.0g/L of a peroxide stabilizer and 5.30 of pH value. Preferably, in the substrate solution I, the alkali solution is 0.4mol/L sodium hydroxide aqueous solution. Preferably, in the first substrate solution, the luminescence enhancer is one of chlorinated metal leaf-ine, histidine/tryptophan/phenylalanine modified metal leaf-ine/chlorinated metal leaf-ine. Preferably, in the first substrate solution, the light-emitting stabilizer is one