A cholinesterase assay reagent and its application

Abstract

The present invention relates to the technical field of in vitro detection reagents, and specifically discloses a cholinesterase assay reagent and its application. The cholinesterase assay reagent of the present invention includes: a reagent one containing potassium ferrocyanide and a buffer and a reagent two containing iodinated butyrylthiocholine and a buffer, wherein the reagent one also includes a surfactant; the reagent two also includes a surfactant and a stabilizer; the stabilizer is dimethyl sulfoxide or dimethylacetamide; the surfactant is one or two of polyoxyethylene dodecyl ether, a fatty alcohol containing 8 EO ethylene oxide groups, polyethylene glycol monolaurate, and 4-nonylphenyl-polyethylene glycol. The cholinesterase assay reagent of the present invention has accurate test results and improved stability: when the bottle is opened at 2-8°C, the result of the assay sample can be continuously stable for 60 days, and during the sealed light-proof storage process at 2-8°C, the result of the assay sample can be continuously stable for 24 months, and the reagent has no color change, which is conducive to storage and use.

Description

Technical Field

[0001] The invention relates to the technical field of in vitro detection reagents, in particular to a cholinesterase determination reagent and application thereof. Background Art

[0002] Cholinesterase (CHE) is an enzyme that catalyzes the hydrolysis of acylcholine or choline esters. It can be divided into two categories according to its substrate specificity, namely true cholinesterase and pseudocholinesterase. True cholinesterase, also known as acetylcholinesterase, is mainly present in the synaptic cleft of cholinergic nerve endings, especially in the wrinkles of the postsynaptic membrane of the motor nerve end plate. It is also present in cholinergic neurons and red blood cells. Pseudocholinesterase (PCHE) exists in serum or plasma. In addition to acting on acetylcholine, it can also act on other choline compounds.

[0003] In clinical practice, the determination of pseudocholinesterase activity is usually used as an important means to assist in the diagnosis of organophosphorus poisoning and the assessment of liver parenchymal cell damage. The existing technology mainly catalyzes butyrylthiocholine through cholinesterase to hydrolyze it into butyric acid and thiocholine, and thiocholine reacts with the indicator 5,5-dithiobis(2-nitrobenzoic acid) (DTNB) to generate a color-developing substance, and the color-developing substance is tested for absorbance at a specified wavelength to obtain the concentration of the hydrolysis product thiocholine, and then the activity of pseudocholinesterase is obtained. However, in actual applications, DTNB has a high blank absorbance and a high reaction background, resulting in a narrow linear range of detection; at the same time, due to the poor stability of DTNB and butyrylthiocholine and the short storage time of liquid reagents, in actual use, it needs to be prepared into dry powder or lyophilized powder reagents for storage, dissolved or re-dissolved when used, and the stability of the reagent is short after opening the lid. The operation is complicated during clinical use and the reagent needs to be replaced frequently.

[0004] Chinese patent CN106680222 discloses a cholinesterase assay reagent and kit, which adopts the method recommended by the German Society of Clinical Chemistry (DGKC) as the basic principle, that is, cholinesterase hydrolyzes butyrylthiocholine to generate thiocholine and butyric acid, and thiocholine reduces yellow potassium ferrocyanide to almost colorless potassium ferrocyanide. The change in absorbance is detected at 400-420nm, and the rate of decrease in absorbance is proportional to the activity of cholinesterase. The technical solution of this patent includes reagent R1 with a pH value of 7.5-8.0 and reagent R2 with a pH value of 3.5-4.5, and the volume ratio of the reagent R1 to the reagent R2 is 5:1; 1L of the reagent R1 is mainly composed of the following components: 1-3g of sodium dihydrogen phosphate, 20-50g of disodium hydrogen phosphate, 1.2-2.4g of trishydroxymethylaminomethane and 0.6-1.2g of potassium ferrocyanide; 1L of the reagent R2 is mainly composed of the following components: 3-9g of 2-(N-morpholinyl)ethanesulfonic acid, 1-6g of sodium chloride, 0.2-1.0g of disodium ethylenediaminetetraacetate, 1-5g of magnesium chloride, 0.2-1g of ProClin300 and 10-20g of butyrylthiocholine iodide; the application of the above technical solution improves the linear range and accuracy of cholinesterase assay reagents and kits. Specifically, the potassium ferrocyanide in the reagent R1 is used as an indicator, sodium dihydrogen phosphate and disodium hydrogen phosphate are used as acid-base buffers, and tris(hydroxymethyl)aminomethane is used as an inhibitor, which mainly inhibits the hydrolysis reaction of choline. The indicator, acid-base buffer and inhibitor cooperate with each other, and the synergistic reagent R2 overcomes the disadvantage of the indicator being too sensitive, which is conducive to expanding the linear range of the cholinesterase assay reagent; magnesium chloride is added to the reagent R2 as a catalyst to accelerate the reaction speed and improve the accuracy of the test; but during the process of sealed light-proof storage at 2~8°C, the blank absorbance of the reagent will decrease, the reactivity of the reagent will decrease, and the color of the reagent will change from colorless to yellow, and the stability of the reagent is a certain problem. And because the substrate iodinated butyrylthiocholine will gradually hydrolyze in an aqueous solution as the placement time increases, it needs to be prepared into a dry powder or lyophilized powder reagent for storage in actual use, and dissolved or redissolved when used, which is inconvenient to use; when prepared into a liquid reagent, the accuracy of the reagent will decrease as the placement time increases, and the stability of the liquid reagent needs to be further improved. Summary of the invention

[0005] In view of the problems of the prior art, the object of the present invention is to provide a stable cholinesterase assay reagent which can effectively assay the activity of cholinesterase.

[0006] In order to achieve this object, the technical solution of the present invention is as follows:

[0007] A cholinesterase assay reagent comprises: a reagent one containing potassium ferrocyanide and a buffer and a reagent two containing butyrylthiocholine iodide and a buffer, wherein the reagent one further comprises a surfactant; the reagent two further comprises a surfactant and a stabilizer; the stabilizer is dimethyl sulfoxide or dimethylacetamide; the surfactant is one or two of polyoxyethylene dodecyl ether, a fatty alcohol containing 8 EO ethylene oxide groups, polyethylene glycol monolaurate, and 4-nonylphenyl-polyethylene glycol.

[0008] The present invention adopts the basic principle of cholinesterase determination recommended by DGKC, and the reaction substrates potassium ferrocyanide and butyrylthiocholine iodide are respectively prepared in reagent one and reagent two, and it is found through research that when a specific surfactant is added to reagent one, and a specific surfactant and a specific stabilizer are added to reagent two, it is unnecessary to add components such as a catalyst and an inhibitor, and the cholinesterase activity in a serum or plasma sample can be quickly determined while overcoming the disadvantages of an overly sensitive indicator and inaccurate testing, and the cholinesterase determination reagent of the present invention has excellent storage stability and is convenient for use.

[0009] In the present invention, polyoxyethylene lauryl ether is EMULGEN109P, fatty alcohol containing 8 EO ethylene oxide groups is ZUSOLAT 1008 / 85 and OXETAL 800 / 85, polyethylene glycol monolaurate is THEST, and 4-nonylphenyl-polyethylene glycol is Nonidet P40.

[0010] In the cholinesterase assay reagent of the present invention, the mass ratio of the potassium ferrocyanide to the butyrylthiocholine iodide is (2-4):(5-20); the mass ratio of the butyrylthiocholine iodide to the stabilizer is (2-10):1.

[0011] In the reagent 1 of the present invention, the concentration of the potassium ferrocyanide is 0.5 g / L to 1.0 g / L, and the concentration of the surfactant is 1.0 g / L to 10.0 g / L;

[0012] In the reagent 2, the concentration of butyrylthiocholine iodide is 5 g / L to 20.0 g / L, the concentration of the surfactant is 1.0 g / L to 10.0 g / L, and the concentration of the stabilizer is 0.5 g / L to 10.0 g / L.

[0013] Preferably, in the reagent 1, the concentration of potassium ferrocyanide is 0.85 g / L, and the concentration of the surfactant is 2.0 g / L to 5.0 g / L;

[0014] In the reagent 2, the concentration of butyrylthiocholine iodide is 10.0 g / L, the concentration of the surfactant is 2.0 g / L to 5.0 g / L, and the concentration of the stabilizer is 1.0 g / L to 5.0 g / L.

[0015] More preferably, in the reagent 1, the concentration of potassium ferrocyanide is 0.85 g / L, and the concentration of the surfactant is 4.0 g / L;

[0016] In the reagent 2, the concentration of butyrylthiocholine iodide is 10.0 g / L, the concentration of the surfactant is 2.0 g / L, and the concentration of the stabilizer is 1.0 g / L.

[0017] The concentration of each substance in the reagent 1 of the present invention can not only meet the requirements of the detection range of different detection instruments but also improve the anti-interference ability of the reagent.

[0018] The concentration of each substance in the reagent 2 of the present invention can ensure the accuracy of detection and the stability of each component.

[0019] When using reagent 1 and reagent 2 of the specific composition of the present invention, the long-term stability of the reagents before mixing can be guaranteed, and after the two are mixed, the components can be properly matched, and the formed reaction system will not affect the accuracy and reaction speed of the test results.

[0020] The cholinesterase assay reagent of the present invention can be configured according to the above-mentioned concentrations of the reagents. When performing the test, reagent one and reagent two are mixed in proportion according to the mass ratio of potassium ferrocyanide and butyrylthiocholine iodide defined above (e.g., the volume ratio of reagent one to reagent two is 4:1).

[0021] In the present invention, the buffer in the reagent 1 is sodium pyrophosphate-sodium dihydrogen phosphate buffer or sodium pyrophosphate-citric acid buffer, and the pH value is 7.4-7.8;

[0022] The buffer in the reagent 2 is 1,4-piperazinediethanesulfonic acid (PIPES) buffer or 2-(N-morpholino)ethanesulfonic acid (MES) buffer, with a pH value of 3.8-4.2;

[0023] The concentration of the buffer component in the buffer in the reagent one is 95-105 mmol / L, and the concentration of the buffer component in the buffer in the reagent two is 10-100 mmol / L.

[0024] Preferably, the surfactant in the reagent 1 is a fatty alcohol containing 8 EO ethylene oxide groups; the surfactant in the reagent 2 is polyoxyethylene lauryl ether;

[0025] Alternatively, the surfactants in the reagents 1 and 2 are both 4-nonylphenyl-polyethylene glycol;

[0026] Alternatively, the surfactant in the reagent one is polyethylene glycol monolaurate; the surfactant in the reagent two is 4-nonylphenyl-polyethylene glycol.

[0027] The fatty alcohol containing 8 EO ethylene oxide groups includes ZUSOLAT 1008 / 85 and OXETAL 800 / 85 in a mass ratio of 1:(1-3), preferably in a mass ratio of 1:1.

[0028] Further preferably, in the present invention, the reagent one comprises: 0.85 g / L potassium ferrocyanide, 2.0 g / L ZUSOLAT1008 / 85, 2.0 g / L OXETAL 800 / 85 and sodium pyrophosphate-citric acid buffer (the concentration of sodium pyrophosphate-citric acid in the reagent one is 100 mmol / L);

[0029] The reagent 2 includes: 10.0 g / L butyrylthiocholine iodide, 1.0 g / L dimethylacetamide, 2.0 g / L EMULGEN109P and MES buffer (the concentration of MES in the reagent 2 is 10.0 mmol / L);

[0030] When used, the reagent 1 and the reagent 2 are mixed in a volume ratio of 4:1.

[0031] The present invention also provides a use of the cholinesterase assay reagent in preparing a cholinesterase assay kit.

[0032] The present invention also provides a cholinesterase assay kit, which comprises the cholinesterase assay reagent.

[0033] The present invention also provides a method for detecting cholinesterase activity, which uses the above-mentioned cholinesterase assay reagent or cholinesterase assay kit for assay.

[0034] The beneficial effects of the present invention are at least:

[0035] 1. The cholinesterase assay reagent prepared by using the surfactant, buffer and stabilizer of the present invention has accurate test results and improved stability: when the bottle is opened and used at 2-8°C, the result of the sample assayed by the reagent can be continuously stable for 60 days, while the stability period of the dry powder reagent in the conventional scheme is 5-15 days; during the sealed and light-proof storage at 2-8°C, the result of the sample assayed by the reagent of the present invention can be continuously stable for 24 months, and the reagent has no color change; while the prior art, such as the method provided by Chinese patent CN106680222, can only be stable for 12 months, and after 12 months, the blank absorbance of the reagent gradually decreases from colorless or light yellow.

[0036] 2. The cholinesterase assay reagent of the present invention can be matched with a calibrator and a quality control product containing cholinesterase to form a kit, which can achieve the accuracy and traceability of the cholinesterase test results and ensure that the test results are continuously under control. DETAILED DESCRIPTION

[0037] The preferred embodiments of the present invention will be described in detail below in conjunction with examples. It should be understood that the following examples are provided only for the purpose of illustration and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0038] The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples are commercially available or prepared according to conventional methods in the art unless otherwise specified.

[0039] Example 1

[0040] This embodiment provides a cholinesterase assay reagent of the present invention.

[0041] The specific composition of reagent 1 is shown in Table 1-1.

[0042] Table 1-1

[0043]

[0044] The specific composition of reagent 2 is shown in Table 1-2.

[0045] Table 1-2

[0046]

[0047] The volume ratio of reagent 1 to reagent 2 during use is 4:1.

[0048] Example 2

[0049] This embodiment provides a cholinesterase assay reagent of the present invention.

[0050] The specific composition of reagent 1 is shown in Table 2-1.

[0051] Table 2-1

[0052]

[0053] The specific composition of reagent 2 is shown in Table 2-2.

[0054] Table 2-2

[0055]

[0056] The volume ratio of reagent 1 to reagent 2 during use is 4:1.

[0057] Example 3

[0058] This embodiment provides a cholinesterase assay reagent of the present invention.

[0059] The specific composition of reagent 1 is shown in Table 3-1.

[0060] Table 3-1

[0061]

[0062] The specific composition of Reagent 2 is shown in Table 3-2.

[0063] Table 3-2

[0064]

[0065] The volume ratio of reagent 1 to reagent 2 during use is 4:1.

[0066] Example 4

[0067] This embodiment provides a cholinesterase assay reagent of the present invention.

[0068] The specific composition of reagent 1 is shown in Table 4-1.

[0069] Table 4-1

[0070]

[0071] The specific composition of Reagent 2 is shown in Table 4-2.

[0072] Table 4-2

[0073]

[0074] The volume ratio of reagent 1 to reagent 2 during use is 4:1.

[0075] Example 5

[0076] This embodiment provides a cholinesterase assay reagent of the present invention.

[0077] The specific composition of reagent 1 is shown in Table 5-1.

[0078] Table 5-1

[0079]

[0080] The specific composition of Reagent 2 is shown in Table 5-2.

[0081] Table 5-2

[0082]

[0083] The volume ratio of reagent 1 to reagent 2 during use is 4:1.

[0084] Comparative Example 1

[0085] This comparative example (based on patent CN1978660A) provides a cholinesterase assay reagent. Compared with the present invention, the reagent has the same reaction principle, and the main components of the reagent both contain potassium ferrocyanide and butyrylthiocholine, but the stabilizer and buffer used in the reagent are different and the reagent does not contain a surfactant. The specific composition is as follows:

[0086] The specific composition of reagent 1 is shown in Table 6-1.

[0087] Table 6-1

[0088]

[0089] The specific composition of Reagent 2 is shown in Table 6-2.

[0090] Table 6-2

[0091]

[0092] Comparative Example 2

[0093] This comparative example provides a cholinesterase assay reagent.

[0094] The components are basically the same as those in Example 1, except that Triton X-100 is used instead of EMULGEN 109P in reagent 2.

[0095] Comparative Example 3

[0096] This comparative example provides a cholinesterase assay reagent.

[0097] The components are substantially the same as those in Example 1, except that glycerol is used instead of dimethylacetamide in reagent 2.

[0098] Comparative Example 4

[0099] This comparative example provides a cholinesterase assay reagent.

[0100] The same stabilizer is used as in Example 2, except that the reagent 1 does not contain a surfactant; the amounts of DMSO and butyrylthiocholine iodide used in the reagent 2 are greater than those in Example 2, and the reagent 2 does not contain the surfactant EMULGEN109P. The specific composition is as follows:

[0101] The specific composition of reagent 1 is shown in Table 7-1.

[0102] Table 7-1

[0103]

[0104] The specific composition of Reagent 2 is shown in Table 7-2.

[0105] Table 7-2

[0106]

[0107] The volume ratio of reagent 1 to reagent 2 during use is 4:1.

[0108] Comparative Example 5

[0109] This comparative example (based on patent CN106018308A) provides a cholinesterase assay reagent, which has a different reaction principle and main components from the present invention, and uses 5,5-dithiobis(2-nitrobenzoic acid) (DTNB) as an indicator.

[0110] The specific composition of reagent 1 is shown in Table 8-1.

[0111] Table 8-1

[0112]

[0113] The specific composition of Reagent 2 is shown in Table 8-2.

[0114] Table 8-2

[0115]

[0116] Experimental Example 1

[0117] This experimental example verifies the test accuracy of the cholinesterase assay reagent of Examples 1-5.

[0118] Specifically, reagent 1 and reagent 2 of Examples 1-5 were used respectively, and the measurement was performed using a fully automatic biochemical analyzer with dual reagent function, and the calibration product was purchased from Biosino Biotechnology Co., Ltd. The specific operation steps of Hitachi 3500 fully automatic biochemical analyzer are shown in Table 9.

[0119] Table 9 Operation steps of fully automatic biochemical analyzer

[0120]

[0121] Wavelength: 405nm, temperature: 37℃, cuvette light path: 1cm.

[0122] The sample is a working calibrator (available from Biosino Biotech Co., Ltd.), which is a composite product. Its main components include 26 routine test items such as cholinesterase, aspartate transferase, alanine transferase, alkaline phosphatase, uric acid, urea, etc. The theoretical concentration of cholinesterase in the product used in this experiment is 4747U / L. The test results are shown in Table 10.

[0123] Table 10 Accuracy test results of the embodiment (unit: U / L)

[0124]

[0125] The accuracy of each embodiment meets the requirements.

[0126] Experimental Example 2

[0127] In this experimental example, the stability of the cholinesterase assay reagents of Examples 1-5 and Comparative Example 5 in the open bottle use state at 2 to 8°C was tested; the stability of the cholinesterase assay reagents of Examples 1-5 and Comparative Example 1-4 in the sealed light-proof storage process at 2 to 8°C was tested.

[0128] Test method: Examples 1-5 and Comparative Examples 1-4 use the method in Experimental Example 1; the test method for Comparative Example 5 is: 5 μL sample + 250 μL reagent 1, pre-incubate for 5 minutes after mixing, add 50 μL reagent 2, read the absorbance A1; read the absorbance A2 after 5 minutes of reaction; the instrument for detecting absorbance will automatically calculate the absorbance change rate per minute (△A / min 样本 ), and give the sample test results.

[0129] Test sample: Biochemical composite quality control product (purchased from Biosino Biotechnology Co., Ltd.), which is a composite product containing 55 routine test items such as cholinesterase, aspartate transferase, alanine transferase, alkaline phosphatase, uric acid, urea, etc. The theoretical concentration of cholinesterase in the product used in this experiment is 8105 U / L.

[0130] Test results (1): When the bottle is opened and used at 2-8°C, measure the absorbance of the sample and reagent blank and compare it with the initial test results. A deviation exceeding ±10% indicates a decrease in the stability of the reagent. The test results are shown in Table 11.

[0131] Table 11 Results of reagent stability test when the bottle is opened

[0132]

[0133] For the reagents of Examples 1-5, within 63 days of being opened and used at 2-8°C, the deviations of the measured sample and reagent blank absorbance from the initial state were all within ±5%, with small fluctuations; for the reagent of Comparative Example 5, as the storage time increased, the measured value decreased and the reagent blank absorbance increased.

[0134] Test results (2): During the storage process at 2-8°C in a sealed and dark place, the absorbance of the sample and reagent blank and the analytical sensitivity were measured and compared with the initial test results. A deviation exceeding ±10% indicates a decrease in the stability of the reagent. The test results are shown in Table 12.

[0135] Table 12 Results of reagent stability test during sealed light-proof storage

[0136]

[0137] The above experimental results show that for Examples 1-5, the blank absorbance of the sample and reagent and the analytical sensitivity were measured at 2-8°C for 24 months in a sealed and dark place, and the initial test results were all within ±10%, indicating that the stability of the reagent was good. For Comparative Examples 1-4, the test results showed a downward trend at 18 months, and the deviation exceeded ±10% at 24 months, indicating that the stability of the reagent decreased.

[0138] Experimental Example 3

[0139] This experimental example verifies the linearity of Examples 1-5.

[0140] The test method is the same as that of Experimental Example 1; the test sample is a high-value sample with cholinesterase added to the serum matrix, and the high-value sample is diluted with physiological saline at a certain ratio before measurement. Test results are shown in Table 13.

[0141] Table 13 Linearity verification results

[0142]

[0143] The linearity of each embodiment is within the range of 700U / L-15000U / L, and the correlation coefficient is greater than 0.990, which can meet clinical needs.

[0144] Experimental Example 4

[0145] This experimental example verifies the analytical sensitivity of Examples 1-5.

[0146] The test method is the same as that of Experimental Example 1; the test sample is a biochemical composite calibrator (purchased from Sinopharm Biotechnology Co., Ltd.), the main components of which are 26 conventional test items including cholinesterase, aspartate transferase, alanine transferase, alkaline phosphatase, uric acid, urea, etc. The test results are shown in Table 14.

[0147] Table 14 Analytical sensitivity verification results

[0148]

[0149] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A cholinesterase assay reagent, characterized in that: include: Reagent 1 containing potassium ferrocyanide and a buffer and reagent 2 containing butyrylthiocholine iodide and a buffer, wherein reagent 1 further comprises a surfactant; reagent 2 further comprises a surfactant and a stabilizer; The surfactant in the reagent 1 is a fatty alcohol containing 8 EO ethylene oxide groups; the surfactant in the reagent 2 is polyoxyethylene lauryl ether; the stabilizer in the reagent 2 is dimethyl sulfoxide or dimethylacetamide; Alternatively, the surfactants in the reagents 1 and 2 are both 4-nonylphenyl-polyethylene glycol; the stabilizer in the reagent 2 is dimethyl sulfoxide; In the reagent 1, the concentration of the potassium ferrocyanide is 0.5 g / L to 1.0 g / L, and the concentration of the surfactant is 1.0 g / L to 10.0 g / L; In the reagent 2, the concentration of butyrylthiocholine iodide is 5 g / L to 20.0 g / L, the concentration of the surfactant is 1.0 g / L to 10.0 g / L, and the concentration of the stabilizer is 0.5 g / L to 10.0 g / L; The mass ratio of the butyrylthiocholine iodide to the stabilizer is (2-10):

1.

2. The cholinesterase assay reagent according to claim 1, characterized in that The mass ratio of the potassium ferrocyanide to the butyrylthiocholine iodide is (2-4):(5-20).

3. The cholinesterase assay reagent according to claim 1, characterized in that In the reagent 1, the concentration of potassium ferrocyanide is 0.85 g / L, and the concentration of the surfactant is 2.0 g / L to 5.0 g / L; In the reagent 2, the concentration of butyrylthiocholine iodide is 10.0 g / L, the concentration of the surfactant is 2.0 g / L to 5.0 g / L, and the concentration of the stabilizer is 1.0 g / L to 5.0 g / L.

4. The cholinesterase assay reagent according to claim 1, characterized in that The buffer in the reagent 1 is sodium pyrophosphate-sodium dihydrogen phosphate buffer or sodium pyrophosphate-citric acid buffer, with a pH value of 7.4-7.8; The buffer in the reagent 2 is 1,4-piperazine diethanesulfonic acid buffer or 2-(N-morpholinyl)ethanesulfonic acid buffer, with a pH value of 3.8-4.2; The concentration of the buffer component in the buffer in the reagent one is 95-105 mmol / L, and the concentration of the buffer component in the buffer in the reagent two is 10-100 mmol / L.

5. The cholinesterase assay reagent according to claim 1, characterized in that: The fatty alcohol containing 8 EO ethylene oxide groups includes ZUSOLAT 1008 / 85 and OXETAL 800 / 85 in a mass ratio of 1:(1-3).

6. Use of the cholinesterase assay reagent according to any one of claims 1 to 5 in the preparation of a cholinesterase assay kit.

7. A cholinesterase assay kit, characterized in that: A cholinesterase assay reagent comprising the reagent according to any one of claims 1 to 5.

8. A method for detecting cholinesterase activity for purposes other than disease diagnosis or treatment, characterized in that: The cholinesterase assay reagent according to any one of claims 1 to 5 or the cholinesterase assay kit according to claim 7 is used for the assay.

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