Chemiluminescence cleaning solution

Abstract

The invention discloses a chemiluminescence cleaning solution, and belongs to the technical field of chemiluminescence immunoassay. The chemiluminescent cleaning solution comprises the following components: a buffer solution, salt, a surfactant, a preservative and a defoaming agent, wherein the surfactant is prepared from Tween 20 and cocoanut fatty acid diethanolamide. According to the invention, the preservative is added into the chemiluminiscence cleaning solution, and multiple preservatives are combined for joint bacteriostasis, so that the concentrated cleaning solution can be stored for 6 months after being diluted, and the detection performance of the reagent is not influenced; various surfactants and high-salt ions are added into the chemiluminescence cleaning liquid, so that the washing effect of the chemiluminescence cleaning liquid is improved.

Description

Technical Field

[0001] This invention relates to the field of chemiluminescence immunoassay technology, and in particular to a chemiluminescence cleaning solution. Background Technology

[0002] The detection steps of chemiluminescence immunoassay are: sample addition, incubation, washing, and detection. The main purpose of washing is to remove unbound immunoreactants and free antigen-antibody conjugates, terminate further antigen-antibody binding, and reduce the influence of non-specific interfering substances on the detection results.

[0003] The cleaning solution is generally composed of buffer solution, salt ions, preservatives, and surfactants. The surfactants and salt ions in the cleaning solution can effectively separate unbound immunoreactants and non-specific interfering substances, ensuring that chemiluminescence immunoassay can be performed normally. The preservatives can ensure the long-term preservation and use of the cleaning solution.

[0004] While the concentrated cleaning solution itself exhibits good corrosion resistance during long-term storage, its resistance becomes problematic when diluted to usable levels. Within just 7-14 days of dilution, significant flocculent matter appears in the solution, leading to decreased accuracy in test results. To ensure accuracy, more frequent maintenance is necessary, requiring servicing of the instrument's cleaning solution tubing every 14 days, significantly increasing the workload. Furthermore, many commercially available cleaning solutions suffer from poor washing effects, high background levels, reduced detection sensitivity, and poor precision; or their washing effect is too strong, resulting in low sensitivity. Summary of the Invention

[0005] The purpose of this invention is to provide a chemiluminescent cleaning solution to solve the problems existing in the prior art.

[0006] To achieve the above objectives, the present invention provides the following solution:

[0007] One of the technical solutions of the present invention is a chemiluminescent cleaning solution, comprising the following components: buffer solution, salt, surfactant, preservative and defoamer;

[0008] The surfactants include Tween 20 and coconut oil fatty acid diethanolamide (6501).

[0009] Furthermore, the concentration of the salt in the chemiluminescent cleaning solution is 100–200 g / L.

[0010] Furthermore, the buffer solution comprises PBS at a concentration of 100 mmol / L.

[0011] The salt includes sodium chloride.

[0012] Furthermore, the concentration of Tween 20 in the chemiluminescent cleaning solution is 2–10 g / L.

[0013] Furthermore, the concentration of the coconut oil fatty acid diethanolamide (6501) in the chemiluminescent cleaning solution is 2-10 g / L.

[0014] Furthermore, the preservatives include Proclin 300 and 5-bromo-5-nitro-1,3-dioxane (BND).

[0015] Furthermore, the concentration of Proclin300 in the chemiluminescent cleaning solution is 10–30 g / L.

[0016] Furthermore, the concentration of 5-bromo-5-nitro-1,3-dioxane (BND) in the chemiluminescent cleaning solution is 1–5 g / L.

[0017] Furthermore, the concentration of the defoamer in the chemiluminescent cleaning solution is 0.5–2 g / L.

[0018] Furthermore, the defoamer includes defoamer 204.

[0019] The second technical solution of the present invention: the application of the above-mentioned chemiluminescent cleaning solution in chemiluminescent immunoassay.

[0020] The present invention discloses the following technical effects:

[0021] (1) The chemiluminescent cleaning solution of the present invention uses two mild nonionic surfactants (Tween20 and 6501) in combination, which enhances the washing effect, reduces the detection background value, and improves the detection signal-to-noise ratio (i.e. sensitivity).

[0022] (2) The chemiluminescent cleaning solution of the present invention uses Proclin 300 and BND in combination, which improves the anti-corrosion effect of the chemiluminescent cleaning solution after dilution into the working solution. Proclin 300 is a commercially available isothiazolinone preservative, which is a mixture of two isothiazolinone compounds and contains a certain amount of magnesium nitrate and magnesium chloride. This type of preservative is a very efficient preservative and has a strong inhibitory effect on various bacteria, molds, yeasts and algae. BND has a broad-spectrum antibacterial effect and can effectively inhibit most bacteria, especially Gram-negative bacteria. The combined use of the two has an excellent antibacterial effect.

[0023] (3) The chemiluminescent cleaning solution of the present invention uses defoamer 204 (a mixture of organic non-silicone polypropylene-based polyether dispersions), which can effectively eliminate foaming and avoid problems such as value jumps and poor precision caused by bubbles.

[0024] (4) This invention achieves the effect that the concentrated cleaning solution can be stored for 6 months after dilution without affecting the detection performance of the reagent by adding preservatives to the chemiluminescent cleaning solution and combining multiple preservatives to inhibit bacteria; by adding multiple surfactants and high salt ions to the chemiluminescent cleaning solution, the washing effect of the chemiluminescent cleaning solution is improved. Detailed Implementation

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0027] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0028] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0029] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0030] It should be noted that any aspects not described in detail in this invention are conventional practices in the field and are not the focus of this invention.

[0031] Example 1

[0032] A chemiluminescent cleaning solution:

[0033] The chemiluminescent cleaning solution was prepared using PBS with a concentration of 100 mmol / L. The chemiluminescent cleaning solution contained sodium chloride at a concentration of 100 g / L, Tween 20 at a concentration of 5 g / L, Proclin 300 at a concentration of 10 g / L, and coconut oil fatty acid diethanolamide (6501) at different concentrations (0 g / L, 2 g / L, 5 g / L, 10 g / L, or 20 g / L).

[0034] Samples at concentrations of 0 and 10 U / mL were tested using a carbohydrate antigen 50 (CA50) assay kit (magnetic microparticle chemiluminescence immunoassay). Each concentration was tested 10 times in duplicate to evaluate the performance of surfactant (6501) at different concentrations. The results are shown in Table 1.

[0035] Table 1 Detection performance of different chemiluminescent cleaning solutions

[0036]

[0037]

[0038] As can be seen from Table 1, the addition of coconut oil fatty acid diethanolamide (6501) to the cleaning solution based on Tween-20 showed a better cleaning effect. The concentration of 2 to 10 g / L can improve the sensitivity (signal-to-noise ratio) of the reagent detection, while the cleaning effect is too strong when the concentration reaches 20 g / L.

[0039] Example 2

[0040] A chemiluminescent cleaning solution:

[0041] The chemiluminescent cleaning solution was prepared using PBS with a concentration of 100 mmol / L. The chemiluminescent cleaning solution contained sodium chloride with a concentration of 100 g / L, Tween 20 with a concentration of 5 g / L, coconut oil fatty acid diethanolamide (6501) with a concentration of 10 g / L, Proclin 300 with a concentration of 10 g / L, and defoamer 204 at different concentrations (0 g / L, 0.5 g / L, 1 g / L, 2 g / L, 4 g / L).

[0042] Samples at concentrations of 0 and 10 U / mL were tested using a carbohydrate antigen 50 (CA50) assay kit (magnetic microparticle chemiluminescence immunoassay). Each concentration was tested 10 times in duplicate to evaluate the performance of defoamer 204 at different concentrations. The results are shown in Table 2.

[0043] Table 2 Detection performance of different chemiluminescent cleaning solutions

[0044]

[0045]

[0046] As can be seen from Table 2, adding defoamer 204 to the cleaning solution can significantly improve the accuracy of detection, especially the accuracy of the background value. The concentration range of 0.5 to 2 g / L is more suitable.

[0047] Example 3

[0048] A chemiluminescent cleaning solution:

[0049] The chemiluminescent cleaning solution was prepared using 100 mmol / L PBS. The solution contained 100 g / L sodium chloride, 5 g / L Tween 20 (TW-20), 10 g / L coconut oil fatty acid diethanolamide (6501), 1 g / L defoamer 204, and different concentrations of preservatives (Proclin 300 and / or 5-bromo-5-nitro-1,3-dioxane (BND)). The specific formulation is shown in Table 3.

[0050] Table 3. Chemiluminescent cleaning solution formulations containing different concentrations of Proclin 300 and BND.

[0051]

[0052] The chemiluminescent cleaning solutions of formulations 1-9 were diluted 10 times and placed in partially sealed containers (simulating the instrument detection environment). The performance of the reagents was observed and tested at different time periods (7 days, 14 days, 28 days, 2 months, 3 months, 4 months, 6 months, and 10 months). The reagent kit used was the carbohydrate antigen 50 (CA50) assay kit (magnetic microparticle chemiluminescent immunoassay).

[0053] Experiments revealed that no turbidity appeared in the formulations at 7 days; at 14 days, formulation 1 showed slight turbidity; at 28 days, both formulations 1 and 2 showed turbidity; at 2 months, formulations 1, 2, 3, and 4 showed turbidity; at 3 months, formulations 1, 2, 3, 4, 5, and 6 showed turbidity; at 6 months, formulations 7, 8, and 9, containing BND preservative, showed no turbidity; and at 10 months, all chemiluminescent cleaning solutions showed turbidity. Performance testing was conducted on the chemiluminescent cleaning solutions that did not show turbidity at 6 months, with a control obtained by diluting the concentrated cleaning solution to the working cleaning solution on the same day. The results were obtained using a carbohydrate antigen 50 (CA50) assay kit (magnetic microparticle chemiluminescent immunoassay). The results are shown in Table 4.

[0054] Table 4 shows the detection performance of chemiluminescent cleaning solutions formulated 1-9.

[0055]

[0056]

[0057] As can be seen from Table 4, adding 1 to 5 g / L of BND to a 10 g / L Proclin 300 solution can significantly improve the anti-corrosion performance of the concentrated cleaning solution after dilution, ensuring that the detection performance of the reagent is not affected for 6 months, and BND has no effect on the detection performance.

[0058] Example 4

[0059] Performance comparison with commercially available cleaning solutions:

[0060] The chemiluminescent cleaning solution prepared in this embodiment is shown in Table 5.

[0061] Table 5. Formulation of Chemiluminescent Cleaning Solution

[0062]

[0063] The two chemiluminescent cleaning solutions of this embodiment were diluted to prepare working solutions (10 times dilution), and two commercially available cleaning solutions (Company A and Company B) were diluted to prepare working solutions according to the instructions. These solutions were then placed in partially sealed containers (simulating the on-machine testing environment) and the following performance tests were conducted:

[0064] (1) Detection performance: The background value, signal-to-noise ratio and precision of the carbohydrate antigen 50 (CA50) assay kit (magnetic microparticle chemiluminescence immunoassay) and the thyroid-stimulating hormone (TSH) assay kit (magnetic microparticle chemiluminescence immunoassay) were compared.

[0065] (2) Anti-corrosion performance: The performance of the reagents was observed and tested at different time periods (1 month, 2 months, 3 months, 4 months and 6 months) using a carbohydrate antigen 50 (CA50) assay kit (magnetic microparticle chemiluminescence immunoassay).

[0066] The results are shown in Tables 6 and 7.

[0067] Table 6. Detection performance of different chemiluminescent cleaning solutions

[0068]

[0069] As can be seen from Table 6, the chemiluminescent cleaning solution of the present invention has a lower background value, a higher signal-to-noise ratio, and better precision than those of Company A and Company B.

[0070] Table 7. Corrosion resistance of chemiluminescent cleaning solutions (after 2 months)

[0071]

[0072]

[0073] Studies have found that Company A's cleaning solution began to become cloudy in the second month, while Company B's cleaning solution began to become cloudy in the first month, and both showed a significant decrease in precision, especially at the zero concentration point. The chemiluminescent cleaning solutions of formulations 1 and 2 of this invention exhibit better corrosion resistance than those of Company A and Company B.

[0074] Comparative Example 1

[0075] Formula 7 is the same as in Example 3, except that coconut oil fatty acid diethanolamide (6501) is replaced with sodium dodecyl sulfate (SDS) of equal concentration.

[0076] Samples at concentrations of 0 and 10 U / mL were tested using a carbohydrate antigen 50 (CA50) assay kit (magnetic microparticle chemiluminescence immunoassay). Each concentration was tested 10 times in duplicate to evaluate the performance of the chemiluminescence cleaning solution. The results are shown in Table 8.

[0077] Table 8 Detection performance of different chemiluminescent cleaning solutions

[0078]

[0079] As can be seen from Table 8, replacing 6501 with SDS of the same concentration resulted in decreased detection precision, reduced signal-to-noise ratio, and a whitening phenomenon in the concentrated mother liquor of the SDS formulation.

[0080] Comparative Example 2

[0081] Formula 7 is the same as in Example 3, except that 5-bromo-5-nitro-1,3-dioxane (BND) is replaced with gentamicin of the same concentration.

[0082] By observing the state of the cleaning solution after dilution (10 times), it was found that in the first month, neither group showed any turbidity; in the second month, the gentamicin group showed turbidity, while the formulation of the present invention did not show turbidity; in the third, fourth, fifth, and sixth months, the gentamicin group showed turbidity, while the formulation of the present invention did not show any turbidity.

[0083] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A chemiluminescent cleaning solution, characterized by comprising: It includes the following components: buffer solution, salt, surfactant, preservative, and defoamer; The surfactants include Tween 20 and coconut oil fatty acid diethanolamide.

2. The chemiluminescent cleaning solution according to claim 1, characterized in that, The concentration of the salt in the chemiluminescent cleaning solution is 100–200 g / L.

3. The chemiluminescent cleaning solution according to claim 1, characterized in that, The buffer solution includes PBS; And / or, the salt includes sodium chloride.

4. The chemiluminescent cleaning solution according to claim 1, characterized in that, The concentration of Tween 20 in the chemiluminescent cleaning solution is 2–10 g / L.

5. The chemiluminescent cleaning solution according to claim 1, characterized in that, The concentration of the coconut oil fatty acid diethanolamide in the chemiluminescent cleaning solution is 2–10 g / L.

6. The chemiluminescent cleaning solution according to claim 1, characterized in that, The preservatives include Proclin 300 and 5-bromo-5-nitro-1,3-dioxane.

7. The chemiluminescent cleaning solution according to claim 5, characterized in that, The concentration of Proclin300 in the chemiluminescent cleaning solution is 10–30 g / L.

8. The chemiluminescent cleaning solution according to claim 5, characterized in that, The concentration of the 5-bromo-5-nitro-1,3-dioxane in the chemiluminescent cleaning solution is 1–5 g / L.

9. The chemiluminescent cleaning solution according to claim 1, characterized in that, The concentration of the defoamer in the chemiluminescent cleaning solution is 0.5–2 g / L.

10. The application of the chemiluminescent cleaning solution according to any one of claims 1 to 9 in chemiluminescent immunoassay.

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