1. A method for detecting the content of water-insoluble glucan is characterized by comprising the following steps:

s1, sample treatment:

s11, adding 1mol/L NaOH solution into a sample to be detected according to a solid-to-liquid ratio of 1: 15-25, and stirring until the NaOH solution is dissolved;

s12, filtering and removing insoluble substances in the step S1, adding 1/5-1/4 GTMAC of the total volume, uniformly mixing, and reacting for 2-4 hours at the temperature of 65-75 ℃;

S2.Ru(bpy)₃ ²⁺modified ga protein:

s21, preparation of Ru (bpy)₃ ²⁺Doped SiO₂；

S22, dispersing the product obtained in the step S21 in a PBS buffer solution containing 5-10% of glutaraldehyde by mass, and stirring for 2-4 h;

s23, dispersing the nano particles obtained by filtering in the step S22 into a PBS (phosphate buffer solution) containing the G alpha a protein, and culturing for 12-24 h at the temperature of 0-4 ℃;

s24, washing the product obtained in the step S23 with a PBS solution, and keeping the product in the PBS buffer solution at the temperature of 0-4 ℃;

s3, preparing the graphene gold nano electrode:

s31, preparing 0.5-1.0 mg/mL GO solution, and adding chloroauric acid into the GO solution to enable the concentration of the chloroauric acid to be 10-50 mu M;

s32, soaking the polished GCE electrode in the solution, and scanning for 8-15 circles within the range of-1.5V-0.5V at the scanning speed of 5-10 mV/s by using a cyclic voltammetry method;

s33, washing the electrode modified in the step S32 with deionized water for 3-5 times;

s4, detecting an object to be detected:

s41, soaking the electrode prepared in the step S3 in PBS (phosphate buffer solution) of the G alpha a protein with the concentration of 0.01-0.05G/mL for 3-5 hours;

s42, cleaning the electrode in the step S41 with a PBS solution, and soaking the electrode in the solution to be detected in the step S1 for 1-3 hours;

s43, cleaning the electrode in the step S42 with a PBS solution, and soaking the electrode in the solution in the step S2 for 0.5-1 h;

and S44, testing the luminous intensity of the electrode in the step S43 in the electrochemical luminescence system.

2. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the solid-to-liquid ratio in step S11 is 1: 20.

3. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the reaction temperature in the step S12 is 70 ℃ and the reaction time is 3 hours.

4. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the pH of the PBS buffer is 7.2.

5. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the concentration of GO in step S31 is 1.0 mg/mL.

6. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the concentration of chloroauric acid in step S31 is 20 μ M.

7. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the scanning speed in step S32 is 10mV/S, and the number of scanning cycles is 10 cycles.

8. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the soaking time in the step S41 is 4 hours.

9. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the soaking time in the step S42 is 1.5 hours.

10. The method for detecting the content of water-insoluble glucan according to claim 1, wherein the soaking time in the step S43 is 1 hour.

Background

Electrochemiluminescence (ECL), also called electrochemiluminescence, is a product of combining chemiluminescence with electrochemistry, and refers to a luminescence phenomenon that some electrically generated substances are generated on the surface of an electrode by applying a certain voltage to perform an electrochemical reaction, and then excited states are formed among the electrically generated substances or among the electrically generated substances and some components in a system through electron transfer, and the excited states return to a ground state. The technology integrates the advantages of high sensitivity of luminescence analysis and controllability of electrochemical potential, and has become one of the research fields of great interest to analytical chemists. The electrochemical luminescence analysis method not only reserves the advantages of high sensitivity, wide linear range, simple equipment, convenient and quick operation, easy realization of automation and the like of the chemical luminescence analysis method, but also has the advantages of strong controllability, good selectivity, capability of providing high-activity luminescent reaction substances, reagent saving and the like of the electrochemical analysis method.

The rapid development of ultrasensitive electrochemiluminescence sensors is mainly attributed to a very good electrochemiluminescence signal amplification strategy. Various nanomaterials, such as carbon nanotubes, graphene, gold nanoparticles, and nanocomposites, have received wide attention as a signal amplifier in the construction of electrochemiluminescence sensors. In particular, graphene is a two-dimensional material with a single atomic thickness, and has remarkable physicochemical properties, such as a large specific surface area and adjustable electronic properties. Graphene has therefore become the most promising electrode material in the field of constructing electrochemiluminescence sensors.

There are many methods for producing graphene, of which the method of reducing Graphene Oxide (GO) obtained by ultrasonic exfoliation is the most effective method. However, since graphene is easily agglomerated in either a dry state or a solution state, practical use of graphene is hindered. The metal nanoparticles were initially introduced into graphene in order to separate graphene sheets and prevent the aggregation of graphene. However, it has now been found that the deposition of metal nanoparticles onto the surface of graphene confers new fields of graphene applications, such as catalysis, magnetic materials, electron conduction, etc. Graphene metal nanocomposites have been applied to ECL biosensors as an electrochemiluminescence signal amplifier, and the resulting sensors have good performance. Therefore, the graphene-gold nanocomposite is also a good material for signal amplification.

The dextran is dextral glucopyranose polymer with molecular formula of (C)₆H₁₀O₅) n is the same as the formula (I). The glucan mainly comprises alpha-dextran and β -dextran. Among them, α -glucan is mainly present in mucus secreted by microorganisms during growth, such as exopolysaccharides produced by bacterial fermentation of sucrose, and is currently one of the best plasma substitutes. The beta-glucan is widely present in microorganisms, plants and even animals, mainly exists in cell walls, has strong induction and activity effects on a foreign body host defense system, and is a good biological response effector with strong activity and low toxic and side effects.

In 2012, the blood of Limulus tridentate and Limulus orientalis was subjected to DNA sequencing by Nippon cantonensis, and a functional region specifically binding to beta-glucan in the fungal cell wall was found. This specific DNA region was named Tachypleus gigas factor G alpha subunit end a (G alpha a).

Examples of deep fungal infections include Candida, Aspergillus, and beta-glucan, a characteristic component of fungal cell walls and recognized by immune cells. Therefore, clinically, the measurement of β -glucan in serum and plasma is used as a criterion for early clinical diagnosis, therapeutic effect, and recovery of fungal infections.

Yeast is an important food and industrial microorganism, and a large amount of insoluble glucan having β -1,3-D as a main chain and β -1,6-D as a side chain is present in the cell wall thereof. The substance has the effects of resisting bacteria and viruses, enhancing the immunity of mammals and the like, is a good biological effect regulator, is widely applied to the industries of food, medicine, cosmetics and the like, and is a hot topic of domestic and foreign research. The water-insolubility of yeast glucan makes it difficult to measure it, and the detection of glucan is difficult.

Disclosure of Invention

The invention aims to overcome the problems in the prior art and provides a method for detecting the content of water-insoluble glucan. The invention utilizes the sandwich-type immunosensor to salinize the beta-glucan quaternary ammonium so as to improve the water solubility of the beta-glucan quaternary ammonium and improve the accuracy of the detection of the beta-glucan.

The purpose of the invention is realized by the following technical scheme:

a method for detecting the content of water-insoluble glucan comprises the following steps:

s1, sample treatment:

s11, adding 1mol/L NaOH solution into a sample to be detected according to a solid-to-liquid ratio of 1: 15-25, and stirring until the NaOH solution is dissolved;

s12, filtering and removing insoluble substances in the step S1, adding 1/5-1/4 GTMAC of the total volume, uniformly mixing, and reacting for 2-4 hours at the temperature of 65-75 ℃;

S2.Ru(bpy)₃ ²⁺modified ga protein:

s21, preparation of Ru (bpy)₃ ²⁺Doped SiO₂；

S22, dispersing the product obtained in the step S21 in a PBS buffer solution containing 5-10% of glutaraldehyde by mass, and stirring for 2-4 h;

s23, dispersing the nano particles obtained by filtering in the step S22 into a PBS (phosphate buffer solution) containing the G alpha a protein, and culturing for 12-24 h at the temperature of 0-4 ℃;

s24, washing the product obtained in the step S23 with a PBS solution, and keeping the product in the PBS buffer solution at the temperature of 0-4 ℃;

s3, preparing the graphene gold nano electrode:

s31, preparing 0.5-1.0 mg/mL GO solution, and adding chloroauric acid into the GO solution to enable the concentration of the chloroauric acid to be 10-50 mu M;

s32, soaking the polished GCE electrode in the solution, and scanning for 8-15 circles within the range of-1.5V-0.5V at the scanning speed of 5-10 mV/s by using a cyclic voltammetry method;

s33, washing the electrode modified in the step S32 with deionized water for 3-5 times;

s4, detecting the object to be detected

S41, soaking the electrode prepared in the step S3 in PBS (phosphate buffer solution) of the G alpha a protein with the concentration of 0.01-0.05G/mL for 3-5 hours;

s42, cleaning the electrode in the step S41 with a PBS solution, and soaking the electrode in the solution to be detected in the step S1 for 1-3 hours;

s43, cleaning the electrode in the step S42 with a PBS solution, and soaking the electrode in the solution in the step S2 for 0.5-1 h;

and S44, testing the luminous intensity of the electrode in the step S43 in the electrochemical luminescence system.

Preferably, the solid-to-liquid ratio in step S11 is 1: 20.

Preferably, the reaction temperature of the step S12 is 70 ℃, and the reaction time is 3 h.

Preferably, the PBS buffer has a pH of 7.2.

Preferably, the concentration of GO in said step S31 is 1.0 mg/mL.

Preferably, the concentration of the chloroauric acid in the step S31 is 20 μ M.

Preferably, in the step S32, the scanning speed is 10mV/S, and the number of scanning turns is 10 turns.

Preferably, the soaking time in the step S41 is 4 h.

Preferably, the soaking time in the step S42 is 1.5 h.

Preferably, the soaking time in the step S43 is 1 h.

The step S1 of the method is to treat the sample mainly to convert the beta-glucan in the sample into quaternary ammonium salt. But also converted into quaternary ammonium salt without influencing the overall molecular configuration of the glucan. The beta-glucan can still be specifically combined with the end a (G alpha a) of the alpha subunit of the factor G of the tachypleus amebocyte. In step S1, GTMAC is glycidyl trimethyl quaternary ammonium hydrochloride.

Step S2 using Ru (bpy)₃ ²⁺A modified ga protein. In step S21, Ru (bpy) is used first₃ ²⁺Modified SiO₂. The specific operation is as follows: first, 1.77mL of surfactant TX-100, 7.5mL of cyclohexane and 1.8mL of co-surfactant n-hexanol were mixed for 30min at room temperature with a mixer. Then, 0.48mL of Ru (bpy) was added₃Cl₂And stirring is continued until the mixture forms a homogeneous water-in-oil microemulsion system. Adding 100 mu L TEOS, then adding 60 mu L ammonia water with the mass percentage of 28-38%, and carrying out hydrolysis reaction. Stirring was continued for 24h, then 100. mu.L (3-aminopropyl) triethoxy silicon (APTES) and 60. mu.L ammonia were added with stirring. After 24h, 25mL of acetone was added to break the emulsion. Centrifuging at 10000r/min for 10min, and collecting the product. Ultrasonic separation with ethanol and waterWashing the core several times to remove the surfactant adsorbed on the surface of the nanoparticles and the adsorbed Ru (bpy)₃Cl₂。

Graphene Oxide (GO) was synthesized from flake graphite by a modified Hummers method. First, 1g of flake graphite and 0.8g of sodium nitrate were weighed and mixed with 23mL of concentrated sulfuric acid for pre-oxidation for 12 hours. Then slowly adding 3.0g of potassium permanganate in an ice bath under stirring, and controlling the sample adding speed to ensure that the temperature is not higher than 30 ℃. It was then transferred to a water bath at about 50 ℃ for 6 h. 3.0g of potassium permanganate is added once again and the reaction is carried out for 6 hours. 50mL of deionized water was added stepwise, the temperature was raised to 98 ℃ and the reaction was maintained at that temperature for 40min, and the mixture was observed to change from tan to yellow. And further adding 30mL of deionized water for dilution, stopping reaction, adding 2mL of hydrogen peroxide solution (30%) for further oxidation, enabling the reaction solution to be mutated into bright yellow, filtering while the reaction solution is hot, repeatedly washing until the reaction solution is neutral, performing 10000rpm centrifugation for 30min after the last washing, pouring out supernate, and performing vacuum drying at 60 ℃ to obtain the graphene oxide.

The processing method of the Glassy Carbon Electrode (GCE) comprises the following specific steps: first, a Glassy Carbon Electrode (GCE) was prepared using 0.3 and 0.05 μm α -Al in this order₂O₃And (5) polishing the powder. The electrode is rinsed with deionized water after each polishing, then ultrasonically treated in deionized water and ethanol for 5min, respectively, and then activated in 0.5M sulfuric acid solution with a scanning range of-1V to 1V and a scanning speed of 100mV s^-1And sweeping until the cyclic voltammograms coincide. The electrode was then used directly for dressing after a simple rinse. Dissolve the obtained GO powder into 0.07M PBS solution with pH of 8.0 to form 1.0mg mL^-1And then a few drops of chloroauric acid solution are added into the GO solution. Using Cyclic Voltammetry (CV) (scan range-1.5V-0.5V, scan speed 10mV s^-1Scan 10 cycles) GO and chloroauric acid were simultaneously reduced to the glassy carbon electrode surface.

The electrochemical luminescence detection system adopts a three-electrode system, and a modified electrode is used as a working electrode. The Pt electrode was used as a counter electrode and AgCl was used as a reference electrode. Ru (bpy)₃ ²⁺An intense electrochemiluminescence signal can be generated in tri-n-propylamine. The electrochemiluminescence solution system of step S4 is a tri-n-propylamine solution.

Compared with the prior art, the invention has the following technical effects:

the method for detecting the content of the water-insoluble glucan adopts the quaternary ammonium salinized glucan, so that the water solubility of the glucan is improved. And the gold nano composite graphene is adopted to expand an electrochemical luminescence signal, so that the detection method with better accuracy and sensitivity is obtained.

Detailed Description

The present invention will be further described in detail with reference to the following specific examples, which are provided for illustration only and are not intended to limit the scope of the present invention. The test methods used in the following examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.

Example 1

A method for detecting the content of water-insoluble glucan comprises the following steps:

s1, sample treatment:

s11, adding 1mol/L NaOH solution into a sample to be detected according to a solid-liquid ratio of 1:15, and stirring until the NaOH solution is dissolved;

s12, filtering and removing insoluble substances in the step S1, adding 1/5 of GTMAC (GTMAC) in total volume, uniformly mixing, and reacting for 2 hours at 75 ℃;

S2.Ru(bpy)₃ ²⁺modified ga protein:

s21, preparation of Ru (bpy)₃ ²⁺Doped SiO₂；

S22, dispersing the product obtained in the step S21 in a PBS buffer solution containing 10% of glutaraldehyde by mass, and stirring for 2 hours;

s23, dispersing the nano particles obtained by filtering in the step S22 into a PBS (phosphate buffer solution) containing the G alpha a protein, and culturing for 24 hours at the temperature of 0-4 ℃;

s24, washing the product obtained in the step S23 with a PBS solution, and keeping the product in the PBS buffer solution at the temperature of 0-4 ℃;

s3, preparing the graphene gold nano electrode:

s31, preparing 1.0mg/mL GO solution, and adding chloroauric acid into the GO solution to enable the concentration of the chloroauric acid to be 10 mu M;

s32, soaking the polished GCE electrode in the solution, and scanning for 8 circles within the range of-1.5V-0.5V at the scanning speed of 5-10 mV/s by using a cyclic voltammetry method;

s33, washing the electrode modified in the step S32 with deionized water for 3 times;

s4, detecting the object to be detected

S41, soaking the electrode prepared in the step S3 in PBS (phosphate buffer solution) of the G alpha a protein with the concentration of 0.01G/mL for 5 hours;

s42, cleaning the electrode in the step S41 with a PBS solution, and soaking the electrode in the solution to be detected in the step S1 for 3 hours;

s43, after the electrode in the step S42 is washed by PBS solution, soaking the electrode in the solution in the step S2 for 0.5 h;

and S44, testing the luminous intensity of the electrode in the step S43 in the electrochemical luminescence system.

Experimental example 2

A method for detecting the content of water-insoluble glucan comprises the following steps:

s1, sample treatment:

s11, adding 1mol/L NaOH solution into a sample to be detected according to a solid-liquid ratio of 1:25, and stirring until the NaOH solution is dissolved;

s12, filtering and removing insoluble substances in the step S1, adding 1/4 of GTMAC (GTMAC) in the total volume, uniformly mixing, and reacting for 4 hours at 65 ℃;

S2.Ru(bpy)₃ ²⁺modified ga protein:

s21, preparation of Ru (bpy)₃ ²⁺Doped SiO₂；

S22, dispersing the product obtained in the step S21 in a PBS buffer solution containing 5% of glutaraldehyde by mass, and stirring for 4 hours;

s23, dispersing the nano particles obtained by filtering in the step S22 into a PBS (phosphate buffer solution) containing the G alpha a protein, and culturing for 12 hours at the temperature of 0-4 ℃;

s24, washing the product obtained in the step S23 with a PBS solution, and keeping the product in the PBS buffer solution at the temperature of 0-4 ℃;

s3, preparing the graphene gold nano electrode:

s31, preparing a 0.5mg/mL GO solution, and adding chloroauric acid into the GO solution to enable the concentration of the chloroauric acid to be 50 mu M;

s32, soaking the polished GCE electrode in the solution, and scanning for 15 circles within the range of-1.5V-0.5V at the scanning speed of 5-10 mV/s by using a cyclic voltammetry method;

s33, washing the electrode modified in the step S32 with deionized water for 3 times;

s4, detecting the object to be detected

S41, soaking the electrode prepared in the step S3 in PBS (phosphate buffer solution) of the G alpha a protein with the concentration of 0.05G/mL for 3 hours;

s42, cleaning the electrode in the step S41 with a PBS solution, and soaking the electrode in the solution to be detected in the step S1 for 1 h;

s43, after the electrode in the step S42 is washed by PBS solution, soaking the electrode in the solution in the step S2 for 1 h;

and S44, testing the luminous intensity of the electrode in the step S43 in the electrochemical luminescence system.

Example 3

A method for detecting the content of water-insoluble glucan comprises the following steps:

s1, sample treatment:

s11, adding 1mol/L NaOH solution into a sample to be detected according to a solid-liquid ratio of 1:20, and stirring until the NaOH solution is dissolved;

s12, filtering and removing insoluble substances in the step S1, adding 1/5 of GTMAC (GTMAC) in the total volume, uniformly mixing, and reacting for 3 hours at 70 ℃;

S2.Ru(bpy)₃ ²⁺modified ga protein:

s21, preparation of Ru (bpy)₃ ²⁺Doped SiO₂；

S22, dispersing the product obtained in the step S21 in a PBS buffer solution containing glutaraldehyde with the mass fraction of 8%, and stirring for 3 hours;

s23, dispersing the nano particles obtained by filtering in the step S22 into a PBS (phosphate buffer solution) containing the G alpha a protein, and culturing for 20 hours at the temperature of 0-4 ℃;

s24, washing the product obtained in the step S23 with a PBS solution, and keeping the product in the PBS buffer solution at the temperature of 0-4 ℃;

s3, preparing the graphene gold nano electrode:

s31, preparing a 1mg/mL GO solution, and adding chloroauric acid into the GO solution to enable the concentration of the chloroauric acid to be 20 mu M;

s32, soaking the polished GCE electrode in the solution, and scanning for 10 circles within the range of-1.5V-0.5V at the scanning speed of 5-10 mV/s by using a cyclic voltammetry method;

s33, washing the electrode modified in the step S32 with deionized water for 3 times;

s4, detecting the object to be detected

S41, soaking the electrode prepared in the step S3 in PBS (phosphate buffer solution) of the G alpha a protein with the concentration of 0.01G/mL for 4 hours;

s42, cleaning the electrode in the step S41 with a PBS solution, and soaking the electrode in the solution to be detected in the step S1 for 1.5 h;

s43, after the electrode in the step S42 is washed by PBS solution, soaking the electrode in the solution in the step S2 for 1 h;

and S44, testing the luminous intensity of the electrode in the step S43 in the electrochemical luminescence system.

Experimental example 1

The gold nano-graphene selenium is obtained by simultaneously depositing graphene oxide and chloroauric acid. First, cyclic voltammetry was performed at 5mM [ Fe (CN) containing 0.1M KCl₆]^4-/[Fe(CN₆)]^3-The deposited electrodes were characterized in solution. Compared with a bare electrode, the modified electrode circuit is greatly improved. The current of the bare electrode is 1.0 x 10^-4A, the current of the modified graphene gold nano rear electrode is changed into 1.4 multiplied by 10^-4A. The electrode modified with the graphene gold nano is soaked in the protein containing the Galpha.a, and the graphene has larger specific surface area and has good adsorption effect on the protein, so that after the Galpha.a protein is adsorbed on the surface of the electrode, the current of the electrode is obviously reduced to 0.4 multiplied by 10^-4A. The change of current in the modification process indicates that the graphene gold nanocomposite is successfully modified by the electrode, and the rapid reduction of the current indicates that the G alpha a protein is adsorbed.

Experimental example 2

In order to explore the composite electricity modified by the graphene gold nanocompositeExtremely stable. Continuous electrochemiluminescence testing was performed at various stages of electrode assembly. The electrochemical luminescence test system is as follows: 5mM [ Fe (CN) containing 0.1M KCl₆]^4-/[Fe(CN₆)]^3-Solution and 0.1M solution of tri-n-propylamine. The scanning range of the electrode is-0.5-1.5V, and the scanning speed is 100 mV/s. The electrode comprises a pair of bare electrodes (GCE), electrodes (GCE-gGNPs) modified with graphene and gold nano composite materials, electrodes (GCE-gGNPs-G alpha a) modified with G-GNPs and adsorbing G alpha a protein, electrodes (GCE-gGNPs-G alpha a-G) soaked in glucan and electrodes (GCE-gGNPs-G alpha a) soaked in G alpha a protein modified with ruthenium. The results of the electrochemiluminescence test on the electrodes at the above-mentioned stages are shown in table 1.

TABLE 1 table of the variation of the electrochemiluminescence intensity during the electrode assembly process

It can be seen from table 1 that the number of cycles gradually increased and decreased with the electrochemical cycle. The electrochemiluminescence signal at each stage decays only slightly. And the bare electrode, the electrode for modifying gGNPs, the stage for adsorbing the G alpha a protein and the stage for re-adsorbing the beta-glucan have almost no corresponding electrochemical signals, and the electrode shows a strong electrochemical luminescence signal only when the G alpha a protein modified with Ru is introduced to the surface of the electrode. And no significant decay of the electrochemiluminescence signal occurs during the continuous electrochemical scan. The electrochemical luminescence biosensor assembled by the method has good stability.

Experimental example 3

Standard beta-glucan solutions were prepared in the series of concentrations of 0.01nM, 0.05nM, 0.1nM, 0.5nM, 1nM, 5nM and 10 nM. The measurement was carried out in accordance with the detection method of example 3, and the electrochemiluminescence intensity at each concentration was measured as shown in Table 2.

TABLE 2 electrochemiluminescence Signal intensities for different concentrations of beta-Glucan

Concentration nM	0.01	0.05	0.1	0.5	1	5	10
								ECL a.u.	345.12	1432.45	2065.78	3179.92	4129.56	4768.33	5467.45

Fitting the data, under the best test condition, the electrochemical luminescence signal of the electrochemical luminescence sensor of the invention increases along with the increase of the concentration of the beta-glucan, and the logarithm of the concentration of the beta-glucan with the electrochemical luminescence intensity in the range of 0.01 nM-10 nM presents a linear relationLinear equation is I_ECL＝1785.8lgC_βG+3843.4, linear correlation coefficient R ═ 0.9987. The standard deviation of the sensor multiplied by 3 times the corresponding intensity of the blank sample is defined as the detection limit, and the detection limit of the sensor is 5.7 pM. Therefore, the invention realizes the ultra-sensitive detection of the beta-glucan.

Experimental example 4

Serum has a chemical composition similar to that of plasma, and the content of beta-glucan in blood is an important index for testing fungal infection in blood. The detection effect of the sensor on the actual sample is determined by a recovery experiment on serum. Standard beta-glucan was added to the serum and the content of beta-glucan was tested by using the method of the above example. The recovery rate is 95-101%. This result demonstrates that the method of the present invention can be applied to the practical detection of β -glucan in blood.

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.