CN111607465A

CN111607465A - Chemiluminescent cleaning solution and preparation method and application thereof

Info

Publication number: CN111607465A
Application number: CN202010517044.7A
Authority: CN
Inventors: 张晓红; 张玉洁; 朱越谭; 曾敏霞
Original assignee: Zhuhai Livzon Diagnostics Inc
Current assignee: Zhuhai Livzon Diagnostics Inc
Priority date: 2020-06-08
Filing date: 2020-06-08
Publication date: 2020-09-01
Anticipated expiration: 2040-06-08
Also published as: CN111607465B

Abstract

The invention provides a chemiluminescent cleaning solution, a preparation method and application thereof, and relates to the technical field of in vitro diagnostic reagents of medical instruments. The cleaning solution has the advantages that the nonionic surfactant and the amphoteric surfactant are used in a combined manner, so that the cleaning solution is good in stability and high in accuracy; different kinds of preservatives are combined, so that the preservative effect is good. The chemiluminescent cleaning solution provided by the invention solves the problems of high price and high detection cost of imported cleaning solution through the selection of specific components, has low cost, and reduces the dependence on imported cleaning solution in China. Meanwhile, the chemiluminescent cleaning solution is suitable for chemiluminescent immunoreaction of an alkaline phosphatase system, and through project verification, the chemiluminescent cleaning solution can be suitable for various chemiluminescent detection projects of alkaline phosphatase systems such as PCT, AMH, dsDNA, CCP and the like.

Description

Chemiluminescent cleaning solution and preparation method and application thereof

Technical Field

The invention relates to the technical field of in-vitro diagnostic reagents for medical instruments, in particular to a chemiluminescent cleaning solution and a preparation method and application thereof.

Background

The chemiluminescence immune reaction comprises the following steps: sample loading, incubation, washing and interpretation. Wherein washing is an important step in determining the outcome of the test. The main purpose of washing is to remove the unbound immunoreactive substance and free conjugate, stop the continuous binding of antigen and antibody, and reduce the influence of non-specific interfering substance on the detection result. The cleaning solution not only provides a proper acid-base environment and electrolyte for immunoreaction, but also removes the immune reactant which is not combined in the reaction process, stops the continuous combination of antigen and antibody, and the surfactant in the cleaning solution can effectively separate the combined immune reactant and non-specific interfering substances, thereby ensuring that the chemiluminescence immunoassay can be normally carried out.

Secondly, the full-automatic chemiluminescence immunoassay analyzer adopts the pipeline, the sample adding needle and the waste liquid needle for repeated utilization, which easily causes the cross contamination of samples and further influences the determination result. Therefore, the effective chemiluminescent cleaning solution has important significance for maintaining pipelines, sample adding needles and waste liquid needles of a full-automatic chemiluminescent immunoassay analyzer and ensuring accurate sample test.

Chemiluminescence can be divided into enzymatic chemiluminescence and non-enzymatic chemiluminescence according to reaction types, in the enzymatic chemiluminescence, an alkaline phosphatase system is one of the more common systems, the reaction speed with a luminescent substrate is high, a correct and reliable result can be provided in a short time, emitted light is glow, the light intensity is relatively stable within 60min, the measurement is easy, the stability is high, and the application is wider.

CN110862881A discloses a special cleaning solution or its diluent for a full-automatic chemiluminescence analyzer, and its preparation method, which mainly comprises inorganic salt, phosphate buffer system, surfactant 1 (one of tween-20 and tween 80), surfactant 2 (one of sodium dodecyl sulfate and sodium metasilicate), preservative (Proclin300 and Bronidox) and water. The cleaning solution employs a relatively high concentration of phosphate, which has been shown to inhibit the activity of alkaline phosphatase. The chemiluminescence reagent of the alkaline phosphatase system has the light emitting principle that alkaline phosphatase catalyzes a substrate adamantane analogue AMPPD to release phosphate radical to form an unstable intermediate AMP-D, the intermediate is decomposed automatically immediately and emits photons, and if phosphate radical with higher concentration exists in the detection reagent system, the catalysis of alkaline phosphatase on the substrate is inhibited, so that the detection reaction is influenced. The Sodium Dodecyl Sulfate (SDS) in surfactant 2 used in the lotion is in powder form, so that the lotion is easy to be inhaled by an operator during weighing, has stimulation on mucosa and upper respiratory tract, eye and skin and is harmful to human body; SDS can destroy the protein structure at high concentration, improper use can cause antigen or antibody structure destruction in immune reaction, influence the detection reaction; SDS solution is easy to generate precipitation at low temperature, and washing effect of washing liquid is influenced.

CN107118865A discloses a cleaning solution, which comprises a phosphate buffer system, inorganic salts, surfactant a (Triton X-100 or tween-20), surfactant B (sodium cholate), and preservative (Proclin300 or Proclin950), and the cleaning solution also adopts phosphate with higher concentration, which is not suitable for alkaline phosphatase chemiluminescence system. The surfactant sodium cholate used in the cleaning solution can dissolve cell membranes, has strong washing capacity, and antigens or antibodies which are specifically combined in the detection process are easily washed away, so that the detection result is inaccurate, and the molecular structure of the sodium cholate contains nitrogen hydrogen bonds with reactivity, so that nonspecific reaction is easily generated in immunoassay. In addition, the preservative Proclin300 or Proclin950 used in the cleaning solution is the same series of preservatives, and cannot achieve a good preservative effect.

CN108546602A discloses a cleaning solution for chemiluminescence immunoassay, which comprises a surfactant, a non-protein protective agent and a buffer, and the cleaning solution has complex components and higher use cost. The amide nonionic surfactant used in the cleaning solution is easy to generate bubbles in the washing oscillation process, so that the liquid level of an instrument is inaccurate to detect, and the phenomenon of value jump is caused. The pH of the cleaning solution was 5.32, which is acidic and not suitable for alkaline phosphatase system. In addition, the preservative Proclin300 used in the cleaning solution is a single preservative, and cannot achieve a good preservative effect.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

It is a first object of the present invention to provide a chemiluminescent cleaning solution for use in an alkaline phosphatase system that alleviates at least one of the problems of the prior art.

The second purpose of the invention is to provide a preparation method of the chemiluminescent cleaning solution.

The invention provides a chemiluminescent cleaning solution applied to an alkaline phosphatase system, which comprises the following components in part by weight:

buffer solution, surfactant, metal ions, preservative and defoaming agent;

the surfactant comprises a nonionic surfactant and an amphoteric surfactant;

the preservative comprises at least two of Proclin-300, BND or propylparaben;

the antifoaming agent includes silicone oil type and/or silicone ether type antifoaming agent.

Further, the concentration of the buffer solution is 100-1000 mM;

the concentration of the surfactant is 0.3% -1.9% w/v;

the concentration of the metal ions is 100-1000 mM;

the concentration of the preservative is 0.3-2.7% w/v;

the concentration of the defoaming agent is 0.05-0.8% w/v.

Further, in the chemiluminescent cleaning solution, the buffer solution comprises a Tris buffer solution;

the nonionic surfactant comprises TritonX-100; amphoteric surfactants include CHAPS;

the metal ions include sodium ions;

the preservative comprises Proclin-300, BND and propylparaben;

the pH of the chemiluminescent cleaning solution is 7.3.

Further, the chemiluminescent washing solution contains 500mM Tris buffer solution, 500mM sodium ions, 0.5w/vCHAPS, 0.5w/v TritonX-100, 0.6% w/v Proclin-300, 0.1% w/v BND, 0.1% w/v propylparaben and 0.1% w/v silicone oil type defoaming agent.

The invention also provides a preparation method of the chemiluminescent cleaning solution, which comprises the steps of uniformly mixing the buffer solution, the surfactant, the metal ions and the preservative, and then adding the defoaming agent to obtain the chemiluminescent cleaning solution;

wherein the surfactant comprises a nonionic surfactant and an amphoteric surfactant;

the preservative comprises at least two of Proclin-300, BND or propylparaben;

Further, the concentration of the buffer solution in the chemiluminescence cleaning solution is 100-1000 mM; the concentration of the surfactant is 0.3% -1.9% w/v; the concentration of the metal ions is 100-1000 mM; the concentration of the preservative is 0.3-2.7% w/v; the concentration of the defoaming agent is 0.05-0.8% w/v.

Further, obtaining a buffer solution in the chemiluminescent cleaning solution, wherein the buffer solution comprises a Tris buffer solution; the nonionic surfactant comprises TritonX-100; amphoteric surfactants include CHAPS; the metal ions include sodium ions; the preservative comprises Proclin-300, BND and propylparaben; the defoaming agent comprises a silicone oil type defoaming agent; the pH of the chemiluminescent cleaning solution is 7.3.

Further, sequentially adding metal ions, a surfactant, a preservative and a defoaming agent into the buffer solution, and then fixing the volume through a solvent to obtain the chemiluminescent cleaning solution;

the solvent comprises water; the final concentration of the buffer solution in the chemiluminescent cleaning solution is 500mM, the final concentration of sodium ions is 500mM, the final concentration of CHAPS is 0.5w/v, the final concentration of TritonX-100 is 0.5v/v, the final concentration of Proclin-300 is 0.6% w/v, the final concentration of BND is 0.1% w/v, the final concentration of propylparaben is 0.1% w/v, and the final concentration of silicone oil type defoaming agent is 0.1% w/v.

Further, after the pH value of the buffer solution is adjusted to 7.0-7.5 by using a solid acid regulator, metal ions, a surfactant and a preservative are sequentially added, and finally, a defoaming agent is added.

Further, the solid acid regulator comprises sulfamic acid and/or boric acid, and the pH of the regulator regulating buffer is 7.3.

The chemiluminescent cleaning solution provided by the invention mainly comprises a buffer solution, a surfactant, metal ions, a preservative and a defoaming agent. The cleaning solution has the advantages that the nonionic surfactant and the amphoteric surfactant are used in a combined manner, so that the cleaning solution is good in stability and high in accuracy; different kinds of preservatives are combined, so that the preservative effect is good. The defoaming agent is added to prevent bubbles from being generated in the process of adding washing liquid or washing, so that the phenomenon of jumping is prevented, and the detection accuracy and precision are improved. The chemiluminescent cleaning solution provided by the invention solves the problems of high price and high detection cost of imported cleaning solution through the selection of specific components, has low cost, and reduces the dependence on imported cleaning solution in China. Meanwhile, the chemiluminescent cleaning solution is suitable for chemiluminescent immunoreaction of an alkaline phosphatase system, and through project verification, the chemiluminescent cleaning solution can be suitable for a plurality of chemiluminescent detection projects of the alkaline phosphatase system such as PCT, AMH, dsDNA, CCP and the like.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

According to an aspect of the present invention, there is provided a chemiluminescent cleaning solution comprising:

buffer solution, surfactant, metal ions, preservative and defoaming agent;

the surfactant comprises a nonionic surfactant and an amphoteric surfactant;

the preservative comprises at least two of Proclin-300, BND or propylparaben;

In the chemiluminescent cleaning solution provided by the invention, the nonionic surfactant and the amphoteric surfactant are jointly used, so that the cleaning solution has good stability, the influence of non-specific interfering substances on the detection result is greatly reduced, and the detection accuracy is improved; different kinds of preservatives are combined, so that the preservative effect is good. The defoaming agent is added to prevent bubbles from being generated in the process of adding washing liquid or washing, so that the phenomenon of jumping is prevented, and the detection accuracy and precision are improved. The chemiluminescent cleaning solution provided by the invention solves the problems of high price and high detection cost of imported cleaning solution by selecting specific components, has low cost, and reduces the dependence on imported cleaning solution in China. Meanwhile, the chemiluminescent cleaning solution is suitable for chemiluminescent immunoreaction of an alkaline phosphatase system, and through project verification, the chemiluminescent cleaning solution can be suitable for various chemiluminescent detection projects of the alkaline phosphatase system, such as PCT, AMH, dsDNA, CCP and other projects.

In the invention, the surfactant can effectively remove nonspecific adsorption substances in the immunoreaction process, and a great amount of experiments prove that the best washing effect can be achieved by adopting the combination of the nonionic surfactant and the amphoteric surfactant, the nonspecific adsorption substances can be completely removed, and the specific binding of protein can not be damaged.

The metal ion inorganic salt can make the cleaning solution have certain ionic strength, thereby protecting the specific binding protein.

In the present invention, the main components of Proclin-series preservatives are 2-methyl-4-isothiazolin-3-one (MCI) and 5-chloro-2-methyl-4-isothiazolin-3-one (CMCI). The two components inhibit cell growth and promote apoptosis by the same principle. Can penetrate cell membrane rapidly, inhibit specific enzyme essential for cell respiration, inhibit cell activity immediately upon contact with microorganism, and has broad antibacterial spectrum and rapid action. BND chemical name is 5-bromo-5-nitro-1, 3-dioxane, reaches the bacteriostatic effect by oxidizing the sulfhydryl in the essential enzyme of microorganism, has broad-spectrum bacteriostatic effect, can effectively inhibit bacteria, yeast and fungi, has good compatibility and stability, low toxicity and is biodegradable. Propyl p-hydroxybenzoate, known as propyl p-hydroxybenzoate, is a colorless crystal or white crystal powder, has no odor, low toxicity, and antibacterial effect on mold and yeast, and can be used as antiseptic in food, cosmetic and medicine. It has low toxicity, is easy to hydrolyze in human body, has antibacterial effect within the pH range of 1-8, and overcomes the disadvantage that common antiseptic is only used under acidic condition. When the three preservatives are preferably combined, the preservatives with different antimicrobial spectra complement each other, so that the antimicrobial range is enlarged, and the optimal bacteriostatic effect is achieved.

In addition, a foam is a dispersion having a plurality of bubbles dispersed in a liquid, the dispersed phase being a gas and the continuous phase being a liquid. When surfactant is added into the system, a layer of surfactant molecules which are arranged in an oriented way is adsorbed on the surface of the air bubble, and when the surfactant molecules reach a certain concentration, the air bubble wall forms a firm film. The surfactant is adsorbed on the gas-liquid interface to cause the surface tension of the liquid surface to be reduced, so that the gas-liquid contact surface is increased, and bubbles are not easy to merge. The bubbles have a relative density much less than that of water and adsorb a layer of surfactant molecules on the surface of the liquid as the rising bubbles permeate the surface. Therefore, the bubble film exposed to the air and adsorbed with the surfactant is different from the bubble film in the solution, and the bubble film is coated with two layers of surfactant molecules to form a bimolecular film, and the adsorbed surfactant has a protective effect on the liquid film. The defoaming agent is to destroy and inhibit the formation of the film, enters the bimolecular oriented film of the foam, and destroys the mechanical balance of the oriented film to break the foam. The ability of silicone to defoam or suppress foam is due to its very low surface tension. The organosilicon compounds (silicone type) interfere with the surface tension of the gas-liquid interface, resulting in defoaming effect. The silicone oil type defoaming agent not only can effectively break the generated foam, but also can remarkably inhibit the foam, prevent the generation of bubbles in the process of adding washing liquor or washing, thereby preventing the occurrence of a jump value phenomenon, improving the detection accuracy and generating a defoaming effect with little use amount, not only having low cost, but also not polluting substances to be defoamed.

In some preferred embodiments, the buffer comprises a Tris buffer, and the use of the Tris buffer as a buffer for the chemiluminescent wash solution is more effective in distinguishing between samples containing analytes and samples not containing analytes in the system of the assay item.

In some preferred embodiments, the nonionic surfactant comprises TritonX-100; the amphoteric surfactant comprises CHAPS.

The surfactant provided by the embodiment does not contain SDS, so that the damage to a body caused by careless inhalation during operation can be effectively avoided, meanwhile, the damage to a protein structure can be avoided, and the normal operation of reaction is ensured. And meanwhile, sodium cholate is not contained, so that inaccurate detection results caused by excessive washing or non-specific reaction are avoided.

In some preferred embodiments, the metal ions include sodium ions.

The addition of the metal ions can provide proper electrolyte for immune reaction, and ensure the normal operation of the reaction. In addition, the source of sodium ions is wide, and the cost of the chemiluminescent cleaning solution provided by the invention can be further reduced by using the sodium ions.

In some preferred embodiments, the pH of the chemiluminescent cleaning solution is 7.3, and when the pH of the chemiluminescent cleaning solution provided by the present invention is 7.3, a suitable acid-base environment can be provided for the alkaline phosphatase chemiluminescent system.

In some preferred embodiments, the concentration of the buffer is 100-1000mM, and for example, but not limited to 100mM, 200mM, 300mM, 400mM, 500mM, 600mM, 700mM, 800mM, 900mM or 1000mM, when the concentration of the buffer is within the above range, the buffer can play a role in buffering, thereby avoiding inhibiting the activity of alkaline phosphatase and ensuring the normal reaction.

Preferably, the concentration of the buffer solution is 500mM, and experiments prove that when the concentration of the buffer solution is 500mM, the luminous value CV verification is minimum, the signal-to-noise ratio is high, which indicates that the cleaning effect of the cleaning solution is better, and the interference substances which are not specifically adsorbed in the immune reaction can be effectively removed.

In some preferred embodiments, the surfactant concentration is 0.3% to 1.9% w/v. For example, but not limited to, 0.3% w/v, 0.4% w/v, 0.5% w/v, 0.8% w/v, 1% w/v, 1.2% w/v, 1.5% w/v, 1.7% w/v, or 1.9% w/v.

The concentration of the surfactant defined in the present embodiment is the total concentration of the nonionic surfactant and the amphoteric surfactant.

Preferably, when the total concentration of the surfactant is 1% w/v, and the concentrations of CHAPS and TritonX-100 are preferably 0.5% w/v through test verification, CV is within 10%, which indicates that the cleaning effect of the cleaning solution is better, and the non-specifically adsorbed interfering substances in the immunoreaction can be effectively removed, the content of the detected substance to be detected in each test is consistent, the repeatability is good, and the signal-to-noise ratio is the highest, the signal-to-noise ratio is a ratio of the signal value of the sample containing the substance to be detected to the signal value of the sample not containing the substance to be detected (or a low-concentration substance to be detected), the larger the ratio is, the larger the difference between the sample containing the substance to be detected and the sample not containing the substance to be detected (or a low-concentration substance to be detected) in the system is, the more effective the separation is achieved, the cleaning effect of the cleaning solution is better, the non-specifically adsorbed interfering substances in the immunoreacti, the content of the substance to be detected in the sample can be reflected more truly.

Preferably, the concentration of the metal ion is 100-1000mM, and may be, for example, but not limited to, 100mM, 200mM, 300mM, 400mM, 500mM, 600mM, 700mM, 800mM, 900mM, or 1000mM, preferably 500 mM.

When the metal ions are in the concentration range, sufficient electrolyte can be provided for the reaction, waste is avoided, and cost is effectively saved.

Preferably, the concentration of the preservative is 0.3% to 2.7% w/v, for example, but not limited to, 0.3% w/v, 0.5% w/v, 0.8% w/v, 1% w/v, 1.2% w/v, 1.5% w/v, 1.8% w/v, 2% w/v, 2.2% w/v, 2.5% w/v or 2.7% w/v. When the concentration of the preservative is within the range, the long-term storage of the chemiluminescent cleaning solution can be ensured.

Preferably the concentration of the preservative is 0.8% w/v, preferably the concentration of Proclin-300 is 0.6% w/v, the concentration of BND is 0.1% w/v, and the concentration of propylparaben is 0.1% w/v. And (3) detecting the blank of the washing liquor to evaluate the bacteriostatic effect of the washing liquor by further screening and limiting the concentration of the preservative. If the washing liquid has a poor bacteriostatic effect and has a bacterial contamination phenomenon, the blank luminous value of the washing liquid is increased, the higher the luminous value is increased, the more serious the bacterial contamination is, and the jump value phenomenon is generated, and experiments prove that the washing liquid can still keep the blank CV of the washing liquid within 5 percent after being placed at 37 ℃ for 6 months, the change of the luminous value is minimum compared with that at 37 ℃ for 6 months, and the washing liquid has the best bacteriostatic effect.

The concentration of the antifoaming agent is preferably 0.05% to 0.8% w/v, and the concentration of the silicone type foaming agent is preferably 0.1% w/v. The cleaning solution added with the defoaming agent has smaller CV and higher signal to noise ratio on the CCP project. The method has the advantages that after the defoaming agent is added into the cleaning liquid, bubbles can be removed or inhibited, the occurrence of a jump value phenomenon in the detection process is effectively reduced, and the precision and the accuracy of reagent detection are improved.

The invention also provides a preparation method of the chemiluminescent cleaning solution, which comprises the steps of uniformly mixing the buffer solution, the surfactant, the metal ions and the preservative to obtain the chemiluminescent cleaning solution;

the preservative comprises at least two of Proclin-300, BND or propylparaben;

The preparation method provided by the invention is simple in process and convenient to operate, and the chemiluminescent cleaning solution can be obtained by mixing the raw materials, so that the preparation method is suitable for actual production, popularization and application.

In some preferred embodiments, metal ions, a surfactant and a preservative are sequentially added into a buffer solution, then an antifoaming agent is added, and then the volume is determined through a solvent to obtain the chemiluminescent cleaning solution. By adding the components to the buffer, the components can be dispersed in a mild environment.

Preferably, the solvent comprises water.

More specifically, the final concentration of the buffer solution in the chemiluminescent cleaning solution is 500mM, the final concentration of sodium ions is 500mM, the final concentration of CHAPS is 0.5w/v, the final concentration of TritonX-100 is 0.5v/v, the final concentration of Proclin-300 is 0.6% w/v, the final concentration of BND is 0.1% w/v, the final concentration of propylparaben is 0.1% w/v, and the final concentration of silicone oil type defoaming agent is 0.1% w/v.

In the specific embodiment of the invention, the order of adding the metal ions, the surfactant and the preservative is not specifically limited, and the order of adding the same substances is not specifically limited, for example, CHAPS and TritonX-100 are both surfactants, and in the actual operation, the CHAPS and TritonX-100 can be added in the order of adding in sequence, or TritonX-100 and CHAPS in sequence, or CHAPS and TritonX-100 can be added simultaneously.

Further, after the pH value of the buffer solution is adjusted to 7.0-7.5, metal ions, a surfactant and a preservative are sequentially added, and the pH value of the buffer solution is preferably adjusted to 7.3;

preferably, the pH of the buffer is adjusted using a solid acid adjusting agent.

The solid acid is used for adjusting the pH value, does not contain SDS and other substances which are easy to absorb, is convenient to prepare and is harmless to operators. In addition, considering that the reagent is in an actual production configuration, concentrated hydrochloric acid and concentrated nitric acid have volatility, and are extremely easy to damage production and configuration personnel and the surrounding environment. Therefore, solid acids are most suitable in combination. It was found by comparison experiments with liquid acids that the chemiluminescent cleaning solution using solid acid for pH adjustment had a greater signal to noise ratio on the CCP project.

Preferably, the solid acid regulator comprises sulfamic acid and/or boric acid, preferably sulfamic acid.

According to the invention, the silicone oil type defoaming agent is added into the chemiluminescent cleaning solution, so that bubbles are prevented from being generated in the process of adding the cleaning solution or washing, the phenomenon of jumping value is prevented, and the detection accuracy and precision are improved.

Preferably, the antifoaming agent comprises one or a combination of two of a silicone oil type antifoaming agent and a silicone ether type antifoaming agent, preferably of the silicone oil type.

In addition, the invention also provides application of the chemiluminescent cleaning solution in a chemiluminescent detection project of an alkaline phosphatase system.

The invention is further illustrated by the following specific examples, which, however, are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

Verification of accuracy, precision, stability and cost

Example 1

The embodiment provides a chemiluminescent cleaning solution, which comprises:

500mM Tris buffer, 0.5% w/v CHAPS, 0.5% w/v Triton X-100, 500mM NaCl, 0.6% w/v Proclin-300, 0.1% w/v BND and 0.1% w/v propylparaben, 0.1% w/v silicone oil type defoamer, pH 7.3, wherein the silicone oil type defoamer is a silicone defoamer sold by Shanghai Biotech under the No. A600465.

Comparative example 1

As described in the CN110862881A patent, the present comparative example provides a chemiluminescent cleaning solution comprising:

sodium chloride	35g
		Potassium chloride	185g
Dodecahydrate and disodium hydrogen phosphate	196.67g
		Potassium dihydrogen phosphate	0.46g
Tween-80	2g
		Sodium dodecyl sulfate	6g
Proclin300	10g
		Bronidox	2g
Dilution factor when in use	20

The pH value of the cleaning solution prepared in the above way is 7.2-8.3.

Comparative example 2

As described in the CN107118865A patent, the present comparative example provides a chemiluminescent cleaning solution comprising:

KH₂PO₄	1.65g
		Na₂HPO₄.12H₂O	67.84g
KCl	180g
		Tween-20	20ml
cholesterol acid sodium salt	10g
		Proclin950	10ml
Dilution factor when in use	20

The pH value of the cleaning solution prepared in the above way is 7.5-8.0.

Comparative example 3

As described in the CN108546602A patent, the present comparative example provides a chemiluminescent cleaning solution comprising:

citric acid-sodium citrate buffer	50mM
		NaCl	150mM
Sodium dodecyl sulfate	0.001％
		Dodecyl polyglycol ether (n2 ═ 23)	0.02％
Coconut diethanolamide	0.005％
		Sucrose	0.5％
(C₁₁H₂₂NO₆P)n1(C₆H₁₀O₃) m1 molecular weight 3 ten thousand (n1: m1 ═ 6:4)	0.3％
		Proclin300	0.09％

The pH value of the cleaning solution prepared in the above way is 5.32, and the use method of the cleaning solution comprises the following steps: 714mL of wash solution +9.286mL of water.

Performance testing

1. Accuracy verification

The cleaning solution of the present invention and the standard cleaning solution of example 1 were diluted according to the ratio indicated in the specification, and then the two cleaning solutions were measured by using a lummel G1200 type full-automatic chemiluminescence immunoassay analyzer, 10 samples were measured for each item, and for each sample measurement item, Procalcitonin (PCT) by the sandwich method, mullerian hormone (AMH), anti-double-stranded DNA antibody (dsDNA) by the indirect method, and anti-cyclic citrullinated peptide antibody (CCP) were selected, and the experimental results are shown in table 1:

table 1 example 1 cleaning solution and standard cleaning solution accuracy test results

As can be seen from Table 1, the deviations of the measured values of the cleaning solutions of example 1 and the standard cleaning solutions in the 4 chemiluminescent products of Procalcitonin (PCT), clamp mullerian hormone (AMH), anti-double-stranded DNA antibody (dsDNA) and anti-cyclic citrullinated peptide antibody (CCP) of the sandwich method were within 5%, which indicates that the cleaning solutions of the present invention have good test accuracy and that the present invention is suitable for AFP, PCT, IL6, TB, AMH, FSH, dsDNA, CCP and the like.

2. Precision verification

The cleaning solution of the invention in example 1, the comparative cleaning solution in comparative example 1, comparative example 2 and comparative example 3 are diluted according to the proportion shown in the specification, and then four cleaning solutions are respectively measured by a LUMIPULSE G1200 type full-automatic chemiluminescence immunoassay analyzer, wherein 1 sample is repeatedly tested 10 times in each item, each test item selects anti-mullerian hormone (AMH) of a sandwich method, and anti-cyclic citrullinated peptide antibody (CCP) of an indirect method are alkaline phosphatase systems, and the test results are shown in Table 2:

table 2 results of testing precision of cleaning solution and comparative cleaning solution of example 1

The precision of the cleaning liquid is within 3%, the precision of the cleaning liquids provided by other comparative examples is more than 7%, the detection value is obviously lower than that of the cleaning liquid provided by the invention, the precision of the cleaning liquid to AMH can be more than 2 times that of the cleaning liquid provided by the invention, the precision of the cleaning liquid to CCP can be more than 3 times that of the cleaning liquid provided by the invention, and the results show that the precision and the reactivity of the cleaning liquid provided by the invention are good, and the cleaning liquid provided by the invention is obviously better than that of the cleaning liquid in an alkaline phosphatase system.

3. Stability verification

The cleaning solution of the invention in example 1 and the comparative cleaning solutions in comparative example 1, comparative example 2 and comparative example 3 were respectively subpackaged into 6 bottles, each bottle has about 1L, and the bottles were respectively placed at room temperature for 0 day, 6 months, 12 months, 18 months, 21 months and 24 months for testing, before testing, the bottles were diluted according to the proportion shown in the specification, then the blank test and CCP project test were respectively carried out on the four cleaning solutions by using a LUMIPULSE G1200 type full-automatic chemiluminescence immunoassay analyzer, and the test was repeated for 10 times, and the test results of each time are shown in tables 3 and 4:

table 3 example 1 cleaning solution and comparative cleaning solution long term stability cleaning solution blank test results

Table 4 example 1 cleaning solution and comparative cleaning solution long term stability CCP test results

The results show that: at room temperature, when the cleaning liquids provided by the comparative examples 1, 2 and 3 are in 18 th month, the blank of the cleaning liquid has a jump value phenomenon, the blank luminous value of the cleaning liquid exceeds 500, which is the phenomenon that the cleaning liquid is infected with bacteria after being stored for a long time, and the blank of the cleaning liquid provided by the invention is stable until 24 th month, and the luminous value is within 300, which shows that the cleaning liquid provided by the invention can still inhibit the growth of bacteria and keep stable within 24 th month. At room temperature, the CV detected by the cleaning liquids provided by the comparative examples 1, 2 and 3 in the project exceeds 10% at the 12 th month, while the CV detected by the cleaning liquid provided by the invention in the project is stable until the 24 th month, and is less than 5%, which indicates that the cleaning liquid provided by the invention in the 24 th month is stable and the stability can reach more than 2 times of that of the control cleaning liquid.

Selection of buffer type

Comparative examples 2 to 1 of example 1

This comparative example differs from example 1 in that the buffer system type is a PBS system, and the other features are the same.

Comparative examples 2 to 2 of example 1

This comparative example differs from example 1 in that the buffer system type was a HEPES system, and the remaining characteristics were the same.

Performance testing

Chemiluminescent cleaning solutions of different buffer types are prepared according to the table 5, the solutions are diluted by 10 times before detection, a LUMIPULSE G1200 full-automatic chemiluminescence immunoassay analyzer is used for measuring the cleaning solutions, 3 samples of Anti-cyclic citrullinated peptide antibodies (Anti-CCP) are selected for each item, the detection is repeated for 10 times, samples with the concentrations of the Anti-cyclic citrullinated peptide antibodies being 0U/mL and 200U/mL are respectively measured for each time, CV is calculated, Anti-cyclic citrullinated peptide antibodies (CCP) are selected for the measuring items, and 0U/mL and 200U/mL CCP are measured for each time. Results the luminescence values (RLU) are shown in table 6:

TABLE 5 chemiluminescent cleaning solutions of different buffer types

TABLE 6 influence of the reactivity of the chemiluminescent wash solution on CCP for different buffer types

The results show that: from the CCP project, the light-emitting value CV verification of the Tris buffer solution is minimum and is within 10 percent; and the signal-to-noise ratio is 449.3, which shows that in the system of the detection item, the sample containing the substance to be detected can be effectively distinguished from the sample without the substance to be detected, so that the buffer solution in the chemiluminescent cleaning solution is Tris (Tris) which is most suitable.

Thirdly, selection of buffer solution concentration

Examples 1-3-1

This example differs from example 1 in that the Tris buffer concentration is 100mM, and the other features are the same.

Examples 1 to 3 and 2

This example differs from example 1 in that the Tris buffer concentration is 1000mM, the remaining characteristics being the same.

Performance testing

Chemiluminescent cleaning solutions with different buffer solution concentrations are prepared according to the table 7, the solutions are diluted by 10 times before detection, a LUMIPULSE G1200 type full-automatic chemiluminescent immunoassay analyzer is used for measuring the cleaning solutions, an Anti-cyclic citrullinated peptide antibody (Anti-CCP) is selected as a measurement item to be repeatedly detected for 10 times, samples with the concentrations of the Anti-cyclic citrullinated peptide antibody being 0U/mL and 200U/mL are measured for each time, and CV and the signal-to-noise ratio of a luminescence value are calculated. The results of the experiment are shown in table 8:

TABLE 7 chemiluminescent cleaning solutions of different buffer concentrations

TABLE 8 influence of the reactivity of the chemiluminescent wash solution on CCP at different buffer concentrations

The results show that: from the CCP project, the light-emitting value CV verification of the Tris buffer solution with the concentration of 500mM is minimum and is within 10 percent; and the signal-to-noise ratio was the highest at 478.8. Therefore, the buffer concentration in the chemiluminescent washing solution is preferably 500 mM.

Fourthly, screening the surfactant in the buffer solution

Comparative example 4-1 of example 1

The embodiment provides a chemiluminescent cleaning solution, which comprises:

the buffer solution system type is Tris buffer solution with the concentration of 500mM, Tween-200.5% of surfactant, 500mM NaCl, 0.6% w/v Proclin-300, 0.1% w/v BND, 0.1% w/v propylparaben and 0.1% w/v silicone oil type defoaming agent with the pH of 7.3, wherein the silicone oil type defoaming agent is the silicone oil defoaming agent sold in Shanghai Bioshop under the No. A600465.

Comparative examples 4 to 2 of example 1

This comparative example differs from example 1 in that the surfactant is Triton X-1000.5% and the remaining characteristics are the same.

Comparative examples 4 to 3 of example 1

This comparative example differs from example 1 in that the surfactant is Brij 0.5% w/v, the remaining characteristics being the same.

Comparative examples 4 to 4 of example 1

This comparative example differs from example 1 in that the surfactant is NP-400.5% w/v, the remaining characteristics being the same.

Comparative examples 4 to 5 of example 1

This comparative example differs from example 1 in that the surfactant is CHAPS 0.5% w/v, the remaining characteristics being the same.

Performance testing

Chemiluminescent cleaning solutions of different surfactants were prepared according to table 9, diluted 10 times before detection, and the cleaning solutions were measured using a lumeal G1200 full-automatic chemiluminescent immunoassay analyzer, and the measurement items selected Anti-cyclic citrullinated peptide antibody (Anti-CCP) for 10 times, samples with concentrations of 0U/mL and 200U/mL for each time of Anti-cyclic citrullinated peptide antibody measurement, and CV and signal-to-noise ratio of the luminescence values were calculated. The results of the experiment are shown in table 10:

TABLE 9 chemiluminescent cleaning solutions of various surfactants

TABLE 10 effect of chemiluminescent purge on CCP reactivity with different surfactants

The results show that: according to empirical values, surfactants with different concentrations are prepared, and from CV verified on CCP project, CV of the surfactants Tween-20, TritonX-100, Brij, NP40 and CHAPS is within 10 percent, which indicates that the surfactants are all stable components; and from the signal-to-noise ratio, triton x-100 and CHAPS are the highest, 412.1 and 411.2, respectively. Therefore, the surfactant in the chemiluminescent cleaning solution is the most suitable surfactant TritonX-100 and CHAPS.

Fifth, the surfactant in the buffer solution is overlapped and screened

Comparative example 5-1 of example 1

This comparative example differs from example 1 in that the surfactant is 0.5% w/v CHAPS, with the same remaining characteristics.

Comparative examples 5 to 2 of example 1

This comparative example differs from example 1 in that the surfactant is TritonX-1000.5% w/v, the remaining characteristics being the same.

Performance testing

Chemiluminescent cleaning solutions of different surfactants were prepared according to table 11, diluted 10 times before detection, and the cleaning solutions were measured using a lumeal G1200 full-automatic chemiluminescent immunoassay analyzer, and the measurement items selected Anti-cyclic citrullinated peptide antibody (Anti-CCP) for 10 times, samples with concentrations of 0U/mL and 200U/mL for each time of Anti-cyclic citrullinated peptide antibody measurement were calculated for CV and signal-to-noise ratio of the luminescence value. The results of the experiment are shown in table 12:

TABLE 11 chemiluminescent cleaning solutions of different surfactants

TABLE 12 effect of chemiluminescent purge on CCP reactivity with different surfactants

The results show that: from the verification of the light-emitting value CV and the verification of the signal-to-noise ratio on the CCP project, the CV is within 10 percent when the surfactants CHAPS and TritonX-100 are used in a superposition mode; the highest signal-to-noise ratio was 458.7. Therefore, the overlapping use effect of CHAPS and TritonX-100 is the best.

Sixthly, screening the concentration of the surfactant

Examples 1 to 6 to 1

This example differs from example 1 in that the surfactant is TritonX-1000.2% w/v, 0.5% w/v CHAPS, and the remaining characteristics are the same.

Examples 1 to 6 and 2

This example differs from example 1 in that the surfactant is TritonX-1000.9% w/v, 0.5% w/v CHAPS, and the remaining characteristics are the same.

Examples 1 to 6 to 3

This example differs from example 1 in that the surfactant is TritonX-1000.5% w/v, 0.1% w/v CHAPS, and the remaining characteristics are the same

Examples 1 to 6 to 4

This example differs from example 1 in that the surfactant is TritonX-1000.5% w/v, 1.0% w/v CHAPS, and the remaining characteristics are the same.

Performance test

Chemiluminescent cleaning solutions with different surfactant concentrations were prepared according to table 13, diluted 10 times before detection, and the cleaning solutions were measured using a lumeal G1200 full-automatic chemiluminescent immunoassay analyzer, with the measurement items selecting Anti-cyclic citrullinated peptide antibodies (Anti-CCP) for 10 times, each time measuring samples with Anti-cyclic citrullinated peptide antibody concentrations of 0U/mL and 200U/mL, respectively, and calculating CV and signal-to-noise ratio of the luminescence values. The results of the experiment are shown in table 14:

TABLE 13 chemiluminescent cleaning solutions of different surfactant concentrations

TABLE 14 effect of chemiluminescent purge on CCP reactivity at different surfactant concentrations

The results show that: as can be seen from the verification of the light-emitting value CV and the verification of the signal-to-noise ratio on the CCP project, the CV is within 10% when the CHAPS concentration of the surfactant is 0.5% w/v and the TritonX-100 concentration is 0.5% w/v; the signal-to-noise ratio was highest and the group 2 signal-to-noise ratios were 447.6. Therefore, the CHAPS concentration is 0.5% w/v and the TritonX-100 concentration is 0.5% w/v, and the effect is best.

Seventh, screening of acid for pH adjustment

Comparative example 7-1 of example 1

This comparative example differs from example 1 in that the pH adjustment is carried out using nitric acid, and the remaining characteristics are the same.

Comparative examples 7 to 2 of example 1

This comparative example differs from example 1 in that the pH adjustment is carried out using hydrochloric acid, and the remaining characteristics are the same.

Performance testing

Chemiluminescent cleaning solutions with PH adjusted by different acids were prepared according to table 15, diluted 10 times before detection, and the cleaning solutions were measured using a lumeal G1200 full-automatic chemiluminescent immunoassay analyzer, and the Anti-cyclic citrullinated peptide antibody (Anti-CCP) was selected for the measurement items and repeatedly detected 10 times, samples with concentrations of 0U/mL and 200U/mL for each measurement were obtained, and CV and signal-to-noise ratio of the luminescence values were calculated. The results of the experiment are shown in table 16:

TABLE 15 chemiluminescent cleaning solution with pH adjusted with different acids

TABLE 16 chemiluminescent wash solutions with pH adjusted with different acids to counter the reactive effect of cyclic citrullinated peptide antibodies

The results show that: the CV of the luminous value of the cleaning solution of which the pH value is adjusted by the three acids is within 10 percent, and the signal to noise ratio of the cleaning solution prepared by the solid acid on the CCP project is larger. Considering that in the actual production configuration of the reagent, concentrated hydrochloric acid and concentrated nitric acid have volatility and are extremely easy to cause damage to production and configuration personnel and the surrounding environment. Therefore, solid acids are most suitable in combination.

Eighthly, screening of preservative types

Comparative example 8-1 of example 1

This comparative example differs from example 1 in that the preservative is Proclin-3000.6% w/v, the remaining characteristics being the same.

Comparative examples 8 to 2 of example 1

This comparative example differs from example 1 in that the preservative was proclin 9500.6% w/v, the remaining characteristics being the same.

Comparative examples 8 to 3 of example 1

This comparative example differs from example 1 in that the preservative is NaN30.2% w/v, the remaining characteristics being the same.

Comparative examples 8 to 4 of example 1

This comparative example differs from example 1 in that the preservative is BND 0.1% w/v, the remaining features being the same.

Comparative examples 8 to 5 of example 1

This comparative example differs from example 1 in that the preservative is methyl paraben 0.1% w/v, the remaining characteristics being the same.

Comparative examples 8 to 6 of example 1

This comparative example differs from example 1 in that the preservative is 0.1% propyl paraben, and the remaining characteristics are the same.

Comparative examples 8 to 7 of example 1

This comparative example differs from example 1 in that the preservative is 0.1% butyl paraben, with the same characteristics.

Performance testing

Chemiluminescent cleaning solutions of different preservatives were prepared according to table 17, and were split into 5 bottles of about 100mL each, and were placed in a 37 ℃ constant temperature incubator for 0 day, 7 days, 1 month, 3 months, and 6 months respectively before detection, diluted 10 times before detection, and cleaning solution blank was determined using a lumip G1200 type full-automatic chemiluminescent immunoassay analyzer, and detection was repeated 10 times, and the mean value and CV were recorded. The results of the experiment are shown in table 18:

TABLE 17 chemiluminescent cleaning solutions with different preservatives

TABLE 18 influence of chemiluminescent cleaning solution on the wash blank value with different preservatives

The results show that: after the cleaning solution is placed in the constant-temperature incubator at 37 ℃ for a period of time, the blank values and CVs of the cleaning solutions 2, 3, 5 and 7 are increased rapidly, which indicates that the cleaning solution is possibly infected with bacteria, and the blank values and CVs of the cleaning solutions 1, 4 and 6 are stable, which indicates that the preservative has a good bacteriostatic effect, so that the preservative Proclin300, BND and propylparaben have the best effect.

Nine, superposition screening of preservative

Examples 1 to 9-1

This example differs from example 1 in that the preservative is Proclin300 at 0.6% w/v, propylparaben at 0.1% w/v, the remaining features being the same.

Examples 1 to 9 and 2

This example differs from example 1 in that the preservative is Proclin300 at 0.6% w/v and BND at 0.1% w/v, the remaining features being the same.

Performance testing

Chemiluminescent cleaning solutions of different preservatives were prepared according to table 19, and were split into 5 bottles of about 100mL each, and were placed in a 37 ℃ constant temperature incubator for 0 day, 7 days, 1 month, 3 months, and 6 months respectively before detection, diluted 10 times before detection, and cleaning solution blank was determined using a lumip G1200 type full-automatic chemiluminescent immunoassay analyzer, and detection was repeated 10 times, and the luminescence mean value and CV were recorded. The results of the experiment are shown in table 20:

TABLE 19 chemiluminescent cleaning solutions with various preservatives

TABLE 20 measurement of washing solution blank values for washing solutions with different preservatives

The results show that: when the preservative is used independently, the blank increase and CV increase of the washing liquid are faster along with the increase of the standing time of the washing liquid, which indicates that the effect of using the preservative independently is good without the addition of the preservative, and the addition of the preservatives Proclin300, BND and propylparaben in the group 3 has the best effect.

Ten, screening of preservative concentration

Examples 1 to 10-1

The embodiment provides a chemiluminescent cleaning solution, which comprises:

500mM Tris buffer, 0.5% w/v CHAPS, 0.5% w/v Triton X-100, 500mM NaCl, 0.05% w/v BND, 0.6% w/v Proclin300, 0.1% w/v propylparaben and 0.1% w/v silicone oil type defoamer, pH 7.3, wherein the silicone oil type defoamer is a silicone defoamer sold by Shanghai Biotech under the code A600465.

Examples 1 to 10 to 2

This example differs from examples 1-10-1 in that the preservative is BND at 0.1% w/v, Proclin300 at 0.6% w/v, propylparaben at 0.1% w/v, and the remaining features are the same.

Examples 1 to 10 to 3

This example differs from examples 1-10-1 in that the preservative is BND at 0.2% w/v, Proclin300 at 0.6% w/v, propylparaben at 0.1% w/v, and the remaining features are the same.

Examples 1 to 10 to 4

This example differs from examples 1-10-1 in that the preservative is BND at 0.1% w/v, Proclin300 at 0.2% w/v, propylparaben at 0.1% w/v, and the remaining features are the same.

Examples 1 to 10 to 5

This example differs from examples 1-10-1 in that the preservative is BND at 0.1% w/v, Proclin300 at 1.5% w/v, propylparaben at 0.05% w/v, and the remaining features are the same.

Examples 1 to 1010 to 6

This example differs from examples 1-10-1 in that the preservative is BND at 0.1% w/v, Proclin300 at 0.6% w/v, propylparaben at 0.05% w/v, and the remaining features are the same.

Performance testing

Chemiluminescent cleaning solutions with different concentrations of preservatives were prepared according to table 21, and were split into 5 bottles of about 100mL each, and were placed in a 37 ℃ constant temperature incubator for 0 day, 7 days, 1 month, 3 months, and 6 months respectively before detection, diluted 10 times before detection, and cleaning solution blank determination was performed on the cleaning solutions using a lumip G1200 type full-automatic chemiluminescent immunoassay analyzer, detection was repeated 10 times, and the luminescence mean value and CV were recorded. The results of the experiment are shown in table 22:

TABLE 21 chemiluminescent cleaning solutions with different concentrations of preservative

TABLE 22 measurement of washing solution blank values of washing solutions at different concentrations of preservative

The results show that: when the preservatives are used together, good antibacterial effects are achieved, when the group 2, namely the concentration of Proclin300 is 0.6%, the concentration of BND is 0.1% and the concentration of propylparaben is 0.1%, is placed at 37 ℃ for 6 months, the blank CV of the lotion can still be kept within 5%, and the change of the luminescence value is minimum compared with that of the lotion at 37 ℃ for 6 months, which indicates that the lotion has the best antibacterial effect, so that the concentration of Proclin300 is 0.6%, the concentration of BND is 0.1% and the concentration of propylparaben is 0.1%, and the effect is the best.

Eleven, screening of antifoaming agent

Comparative example 11-1 of example 1

This comparative example differs from example 1 in that the defoamer is not included and the remaining characteristics are the same.

Examples 1 to 11-1

This example differs from example 1 in that the antifoam is a 0.1% w/v silyl ether antifoam with the same remaining characteristics.

Examples 1 to 11 and 2

This example differs from example 1 in that the antifoam is a silicone oil type antifoam of 0.05% w/v, the remaining characteristics being the same.

Examples 1 to 11 to 3

This example differs from example 1 in that the antifoam is a silicone oil type antifoam of 0.8% w/v, the remaining characteristics being the same.

Performance testing

Chemiluminescent cleaning solutions with different antifoaming agents were prepared according to table 23, diluted 10 times before detection, and the cleaning solutions were measured using a lumeal G1200 full-automatic chemiluminescent immunoassay analyzer, and the Anti-cyclic citrullinated peptide antibody (Anti-CCP) was selected for the measurement items and repeatedly detected 10 times, and each time samples with concentrations of 0U/mL and 200U/mL of Anti-cyclic citrullinated peptide antibody were measured, and CV and signal-to-noise ratio of the luminescence values were calculated. The results of the experiment are shown in table 24:

TABLE 23 chemiluminescent cleaning solutions of various defoamers

TABLE 24 chemiluminescent wash solutions of various antifoam agents to combat the reactive effect of cyclic citrullinated peptide antibodies

The results show that: the cleaning solution added with the defoaming agent has smaller CV and higher signal to noise ratio on the CCP project. The method has the advantages that after the defoaming agent is added into the cleaning liquid, bubbles can be removed or inhibited, the occurrence of a jump value phenomenon in the detection process is effectively reduced, and the precision and the accuracy of reagent detection are improved. Wherein, the cleaning solution of 0.1 percent w/v silicone oil type defoaming agent is added, the CV is minimum, and the signal-to-noise ratio is highest. Therefore, from the viewpoint of compatibility, 0.1% w/v of the silicone oil type defoaming agent is most suitable.

As can be seen from comparison of experimental data of each set of examples, the synergistic effect of the buffer, the surfactant, the pH acid and the preservative enables 500mM Tris buffer, 0.5% w/v CHAPS, 0.5% w/v Triton X-100, 500mM NaCl, 0.6% w/v Proclin-300, 0.1% w/v BND, 0.1% w/v propylparaben and 0.1% w/v silicone oil type antifoaming agent, and the optimal condition of the chemiluminescent cleaning solution of the present invention is that solid acid sulfamic acid is used to adjust the pH to 7.3. The buffer solution can provide a buffer environment for the detection system, so that the solution maintains a relatively stable pH, but does not participate in the detection reaction.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A chemiluminescent cleaning solution for use in an alkaline phosphatase system, the chemiluminescent cleaning solution comprising:

buffer solution, surfactant, metal ions, preservative and defoaming agent;

the surfactant comprises a nonionic surfactant and an amphoteric surfactant;

the preservative comprises at least two of Proclin-300, BND or propylparaben;

2. The chemiluminescent cleaning solution according to claim 1, wherein the buffer solution has a concentration of 100-1000 mM;

the concentration of the surfactant is 0.3% -1.9% w/v;

the concentration of the metal ions is 100-1000 mM;

the concentration of the preservative is 0.3-2.7% w/v;

the concentration of the defoaming agent is 0.05-0.8% w/v.

3. A chemiluminescent cleaning solution according to claim 2 wherein the chemiluminescent cleaning solution is a solution,

the buffer comprises Tris buffer;

the nonionic surfactant comprises TritonX-100;

amphoteric surfactants include CHAPS;

the metal ions include sodium ions;

the preservative comprises Proclin-300, BND and propylparaben;

the pH of the chemiluminescent cleaning solution is 7.3.

4. A chemiluminescent cleaning solution according to claim 3 wherein,

the chemiluminescence cleaning solution contains 500mM Tris buffer solution, 500mM sodium ions, 0.5w/v CHAPS, 0.5w/v Triton X-100, 0.6% w/v Proclin-300, 0.1% w/v BND, 0.1% w/v propylparaben and 0.1% w/v silicone oil type defoaming agent.

5. The method for preparing a chemiluminescent cleaning solution according to any one of claims 1 to 4 wherein the chemiluminescent cleaning solution is obtained by first uniformly mixing a buffer solution, a surfactant, metal ions and a preservative, and then adding a defoaming agent;

the preservative comprises at least two of Proclin-300, BND or propylparaben;

6. The production method according to claim 5, wherein the chemiluminescent cleaning solution is obtained,

the concentration of the buffer solution is 100-1000 mM;

the concentration of the surfactant is 0.3% -1.9% w/v;

the concentration of the metal ions is 100-1000 mM;

the concentration of the preservative is 0.3-2.7% w/v;

the concentration of the defoaming agent is 0.05-0.8% w/v.

7. The production method according to claim 6, wherein the chemiluminescent cleaning solution is obtained,

the buffer comprises Tris buffer;

the nonionic surfactant comprises TritonX-100;

amphoteric surfactants include CHAPS;

the metal ions include sodium ions;

the preservative comprises Proclin-300, BND and propylparaben;

the defoaming agent comprises a silicone oil type defoaming agent;

the pH of the chemiluminescent cleaning solution is 7.3.

8. The preparation method according to claim 7, characterized in that metal ions, a surfactant, a preservative and a defoaming agent are added into the buffer solution in sequence, and then the volume is determined by a solvent to obtain the chemiluminescent cleaning solution;

the solvent comprises water;

the final concentration of the buffer solution in the chemiluminescent cleaning solution is 500mM, the final concentration of sodium ions is 500mM, the final concentration of CHAPS is 0.5w/v, the final concentration of TritonX-100 is 0.5v/v, the final concentration of Proclin-300 is 0.6% w/v, the final concentration of BND is 0.1% w/v, the final concentration of propylparaben is 0.1% w/v, and the final concentration of silicone oil type defoaming agent is 0.1% w/v.

9. The method according to any of claims 5 to 8, wherein the pH of the buffer is adjusted to 7.0 to 7.5 using a solid acid regulator, and then the metal ions, the surfactant, the preservative and the antifoaming agent are sequentially added.

10. The method of claim 9, wherein the solid acid regulator comprises sulfamic acid and/or boric acid, and the pH of the regulator adjustment buffer is 7.3.