CN116735570A - Method for measuring concentration of vancomycin drug by chemical photoparticulate reagent - Google Patents
Method for measuring concentration of vancomycin drug by chemical photoparticulate reagent Download PDFInfo
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- 108010059993 Vancomycin Proteins 0.000 title claims abstract description 65
- 229960003165 vancomycin Drugs 0.000 title claims abstract description 65
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- 239000000427 antigen Substances 0.000 claims description 6
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Abstract
The invention belongs to the technical field of biological detection, and discloses a method for measuring vancomycin drug concentration by using a chemical light microparticle reagent, which is used for preparing a chemical light microparticle reagent kit; taking 1-5 mL of a sample to be detected, centrifuging for 7min at a centrifugation speed of 3500-5000 rpm, and taking supernatant to obtain serum or plasma; and detecting serum or plasma by using the kit, and obtaining the corresponding vancomycin drug concentration. The invention combines the chemiluminescence detection technology and the biotin-streptavidin system for the first time to realize the purpose of quantitatively detecting the content of the vancomycin drug in the human serum sample, ensures the detection sensitivity, has no pollution, strong specificity and simple operation, has low pretreatment requirement on the sample, can rapidly detect a large number of samples in a high-flux way, and provides a more accurate, precise, convenient, quick and simple method for clinically detecting the concentration of the vancomycin drug in the human serum.
Description
Technical Field
The invention belongs to the field of biological detection, and particularly relates to a method for measuring vancomycin drug concentration by using a chemical light microparticle reagent.
Background
Vancomycin belongs to glycopeptide antibiotics, and hydrochloride thereof is clinically applied. Vancomycin has powerful bactericidal effect on gram-positive cocci and part of gram-positive bacilli (e.g., diphtheria bacillus, clostridium). The antibacterial activity action mechanism of vancomycin is mainly to inhibit bacterial cell wall synthesis, and the vancomycin can also change the permeability of bacterial cell membranes and influence the synthesis of RNA, so that the product has a powerful bactericidal effect. The effective blood concentration of vancomycin is 5.0-10.0mg/L, and when the blood concentration is more than 10.0mg/L, the toxic and side effects are enhanced. Adverse reactions are often referred to as "red neck syndrome", manifested by erythema, flushing of the face and neck and even the chest, drug fever, hypotension, and even shock-like reactions; and has nephrotoxicity and ototoxicity. Because the vancomycin treatment window is narrow and the metabolic process in the body has larger individual difference, the detection of the drug concentration in the blood is an important reference for adjusting the dosage and reducing adverse reaction, and has great significance in clinical medication. The current common methods for detecting vancomycin therapeutic drugs are as follows: ultraviolet spectrophotometry, homogeneous immunoassay, high performance liquid chromatography, liquid chromatography-mass spectrometry, and the like. Among the numerous methods, liquid chromatography is the most accurate method for determining vancomycin. The prior method has the problems of complex pretreatment, inaccurate qualitative and quantitative analysis, high cost, long analysis time, high technical requirements on experimenters and the like.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a method for measuring the concentration of vancomycin by using a chemical photoparticulate reagent.
The invention is realized by the method for measuring the concentration of vancomycin drug by using a chemical photoparticulate reagent, which is characterized by comprising the following steps:
s1: preparing a chemical photoparticle kit;
s2: taking 1-5 mL of a sample to be detected, centrifuging for 7min at a centrifugation speed of 3500-5000 rpm, and taking supernatant to obtain serum or plasma;
s3: and detecting serum or plasma by using the kit, and obtaining the corresponding vancomycin drug concentration.
Further, the kit in S1 specifically includes:
r1 reagent, R2 reagent, sample treatment fluid, magnetic separation reagent, cleaning fluid, eluent, calibrator and quality control product;
the reagent R1 is prepared from a biotinylated vancomycin monoclonal antibody A and a reagent diluent R1, the reagent R2 is prepared from an alkaline phosphatase-marked vancomycin monoclonal antibody B and a reagent diluent R2, the magnetic separation reagent is prepared from commercial streptavidin magnetic beads and a magnetic separation reagent diluent, and the calibrator and quality control materials are prepared from diluents containing different concentrations of vancomycin antigens and proteins; the chemiluminescent substrate solution needed in the experimental process is substrate solution for alkaline phosphatase catalytic luminescence.
Further, the working concentration of the biotinylated vancomycin monoclonal antibody A conjugate in the R1 reagent is 0.5-2.5 mug/mL; the pH value of the R1 reagent diluent is 7.5-8.0.
Further, the components of the R1 reagent diluent include: tris, working concentration is 12.0-12.7 g/L; sodium azide with the working concentration of 0.48-0.6 g/L; sodium chloride with the working concentration of 5.7-6.0 g/L; bovine serum albumin with a working concentration of 9-10 g/L; methyl cellulose with working concentration of 0.7-1.0 g/L; the balance being purified water.
Further, the working concentration of the vancomycin monoclonal antibody B conjugate marked by alkaline phosphatase in the R2 reagent is 0.7-1.5 mug/mL.
Further, the pH value of the R2 reagent diluent is 7.7-8.0, and the method comprises the following steps: tris, working concentration is 12.0-14.3 g/L; sodium azide with the working concentration of 1.60-2.50 g/L; sodium chloride with the working concentration of 3.7-6.0 g/L; magnesium chloride with the working concentration of 0.5-0.5 g/L; zinc chloride with working concentration of 0.03-0.1 g/L; bovine serum albumin with a working concentration of 5-10 g/L; the working concentration of the AP enzyme protective agent is 400-500 g/L; the balance being purified water.
Further, the sample treatment solution is an aqueous solution of one or more than two of acetate buffer solution, acetonitrile and methanol, wherein the content of ammonium acetate in the acetate buffer solution is 0.1-0.3 per mill, the content of methanol is 5-15%, and the content of acetonitrile is 5-15%.
Further, the cleaning liquid is methanol or acetonitrile or an aqueous solution of two mixtures.
Further, the eluent is an aqueous solution of one or more than two of acetate buffer solution, acetonitrile and methanol, wherein the content of ammonium acetate in the acetate buffer solution is 0.05-0.15 per mill, the content of methanol is 0-15%, and the content of acetonitrile is 70-90%.
Further, the step S3 specifically includes:
(1) Taking serum or plasma, transferring 1.0ml, adding into a sample bottle, and placing into an automatic sampler to sample 100ul for quantitative detection by liquid chromatography;
(2) Respectively adding an R1 reagent and an R2 reagent into a detection tube, and mixing;
(3) (37+ -0.5) deg.C for 15min;
(4) Adding 25 mu L of magnetic particle reagent into the detection tube;
(5) (37+ -0.5) deg.C for 5min;
(6) Adding 600 mu L of cleaning liquid into the detection tube, and uniformly mixing;
(7) Magnetically separating the supernatant to remove unreacted materials;
(8) Repeating the steps (6) - (7) twice;
(9) Adding 150 mu L of luminous substrate into a detection tube, and uniformly mixing for 3s;
(10) Measuring relative luminescence intensity (RLU) after ALP catalyzes luminescence of the substrate;
RLU and vancomycin antigen concentration are positively correlated in a certain range, and the vancomycin content of a sample to be detected is read from a standard curve by an interpolation method.
Further, the method comprises the steps of:
s1: reagent kit for preparing chemical photoparticulate
1.1 selecting suitable chemical reagents, including particulates and other reagents;
1.2 mixing the particles and other reagents according to a certain proportion to prepare; forming a kit;
s2: taking 1-5 mL of a sample to be detected, centrifuging for 7min at a centrifugation speed of 3500-5000 rpm, and taking supernatant to obtain serum or plasma;
2.1 preparing a sample comprising serum or plasma;
2.2, transferring the sample into a centrifuge tube, and performing centrifugal operation, wherein the centrifugal speed is 3500-5000 rpm, and the centrifugal time is 7min;
2.3 taking supernatant to obtain serum or plasma samples;
s3: detecting serum or plasma by using the kit, and obtaining the corresponding vancomycin drug concentration; 3.1, adding serum or plasma samples to be detected and a reagent into the kit, and forming a fluorescent signal after reaction;
3.2, detecting and quantitatively analyzing fluorescent signals by using instruments such as a chemiluminescent instrument and the like to obtain the corresponding concentration of the vancomycin drug.
In combination with the technical scheme and the technical problems to be solved, the technical scheme to be protected has the following advantages and positive effects:
in the method for measuring the concentration of the vancomycin drug by using the chemical light microparticle reagent, the aim of quantitatively measuring the content of the vancomycin drug in a human serum sample is realized by combining a chemiluminescence detection technology with a biotin-streptavidin system for the first time, the detection sensitivity is ensured, the reaction system has no pollution, strong specificity and simple operation, the pretreatment requirement on the sample is low, and a large number of samples can be rapidly and high-flux detected, so that a more accurate, precise, convenient, quick and simple method is provided for clinically measuring the concentration of the vancomycin drug in human serum.
The method for measuring the concentration of the vancomycin drug by using the chemical photoparticulate reagent has the technical effects and advantages as follows:
1) Improving the accuracy and sensitivity of drug determination: by preparing the chemical light microparticle kit, the detection and quantitative analysis of fluorescent signals can be carried out by using instruments such as a chemiluminescent instrument, so that the accuracy and the sensitivity of drug determination are improved.
2) The detection efficiency is improved: the method is in modularized design, is simple in detection operation, can quickly obtain results, and improves detection efficiency.
3) The detection cost is reduced: the kit has low preparation cost, is simple and convenient to operate, and can reduce detection cost.
4) Improving the purity of the sample: through adjusting centrifugal parameters, impurities and precipitates in the sample can be removed better, and the purity of the sample is improved, so that the detection accuracy and sensitivity are improved.
5) Convenient and practical: the method does not need complex instruments and technology, has the characteristics of simplicity, convenience and practicability, and can be widely applied to the fields of medicine quality control, clinical detection and the like.
6) The quantitative analysis can be carried out: the method can quantitatively analyze the concentration of the vancomycin drug and can provide accurate data support for drug research and development, production and medical diagnosis.
7) The reliability is high: the method has accurate and reliable measurement result and high accuracy and stability.
8) Environmental protection and energy saving: the method does not need to use harmful chemical reagents, and has the advantages of environmental protection and energy saving.
In conclusion, the method has the advantages of simplicity and convenience in operation, high sensitivity, high accuracy, low cost, high sample purity, capability of quantitative analysis, high reliability, environmental protection, energy conservation and the like, and can meet the requirements of the fields of medicine quality control, clinical detection and the like.
Drawings
FIG. 1 is a flow chart of a method for determining the concentration of vancomycin drug by using the chemical photoparticulate reagent provided in the embodiment of the invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
As shown in fig. 1, an embodiment of the present invention provides a method for determining vancomycin drug concentration by using a chemical photoparticulate reagent, the method comprising:
s1: preparing a chemical photoparticle kit;
s2: taking 1-5 mL of a sample to be detected, centrifuging for 7min at a centrifugation speed of 3500-5000 rpm, and taking supernatant to obtain serum or plasma;
s3: and detecting serum or plasma by using the kit, and obtaining the corresponding vancomycin drug concentration.
The kit in S1 specifically comprises:
r1 reagent, R2 reagent, sample treatment fluid, magnetic separation reagent, cleaning fluid, eluent, calibrator and quality control product;
the reagent R1 is prepared from a biotinylated vancomycin monoclonal antibody A and a reagent diluent R1, the reagent R2 is prepared from an alkaline phosphatase-marked vancomycin monoclonal antibody B and a reagent diluent R2, the magnetic separation reagent is prepared from commercial streptavidin magnetic beads and a magnetic separation reagent diluent, and the calibrator and quality control materials are prepared from diluents containing different concentrations of vancomycin antigens and proteins; the chemiluminescent substrate solution needed in the experimental process is substrate solution for alkaline phosphatase catalytic luminescence.
The working concentration of the biotinylated vancomycin monoclonal antibody A conjugate in the R1 reagent is 0.5-2.5 mug/mL; the pH value of the R1 reagent diluent is 7.5-8.0.
The components of the R1 reagent diluent comprise: tris, working concentration is 12.0-12.7 g/L; sodium azide with the working concentration of 0.48-0.6 g/L; sodium chloride with the working concentration of 5.7-6.0 g/L; bovine serum albumin with a working concentration of 9-10 g/L; methyl cellulose with working concentration of 0.7-1.0 g/L; the balance being purified water.
The working concentration of the vancomycin monoclonal antibody B conjugate marked by alkaline phosphatase in the R2 reagent is 0.7-1.5 mug/mL.
The pH value of the R2 reagent diluent is 7.7-8.0, and the method comprises the following steps: tris, working concentration is 12.0-14.3 g/L; sodium azide with the working concentration of 1.60-2.50 g/L; sodium chloride with the working concentration of 3.7-6.0 g/L; magnesium chloride with the working concentration of 0.5-0.5 g/L; zinc chloride with working concentration of 0.03-0.1 g/L; bovine serum albumin with a working concentration of 5-10 g/L; the working concentration of the AP enzyme protective agent is 400-500 g/L; the balance being purified water.
The sample treatment solution is an aqueous solution of one or more than two of acetate buffer solution, acetonitrile and methanol, wherein the content of ammonium acetate in the acetate buffer solution is 0.1-0.3 per mill, the content of methanol is 5-15%, and the content of acetonitrile is 5-15%.
The cleaning liquid is methanol or acetonitrile or water solution of two mixtures.
The eluent is an aqueous solution of one or more than two of acetate buffer solution, acetonitrile and methanol, wherein the content of ammonium acetate in the acetate buffer solution is 0.05-0.15 per mill, the content of methanol is 0-15%, and the content of acetonitrile is 70-90%.
The step S3 specifically comprises the following steps:
(1) Taking serum or plasma, transferring 1.0ml, adding into a sample bottle, and placing into an automatic sampler to sample 100ul for quantitative detection by liquid chromatography;
(2) Respectively adding an R1 reagent and an R2 reagent into a detection tube, and mixing;
(3) (37+ -0.5) deg.C for 15min;
(4) Adding 25 mu L of magnetic particle reagent into the detection tube;
(5) (37+ -0.5) deg.C for 5min;
(6) Adding 600 mu L of cleaning liquid into the detection tube, and uniformly mixing;
(7) Magnetically separating the supernatant to remove unreacted materials;
(8) Repeating the steps (6) - (7) twice;
(9) Adding 150 mu L of luminous substrate into a detection tube, and uniformly mixing for 3s;
(10) Measuring relative luminescence intensity (RLU) after ALP catalyzes luminescence of the substrate;
RLU and vancomycin antigen concentration are positively correlated in a certain range, and the vancomycin content of a sample to be detected is read from a standard curve by an interpolation method.
As an optimization scheme of the embodiment of the invention, the specific refinement steps are as follows:
s1: reagent kit for preparing chemical photoparticulate
1.1 selection of suitable chemical reagents including microparticles and other reagents such as fluorescein and the like.
1.2 mixing the microparticles and other reagents according to a certain proportion to prepare the kit.
S2: taking 1-5 mL of sample to be detected, centrifuging for 7min at the centrifugal speed of 3500-5000 rpm, and taking supernatant to obtain serum or plasma.
2.1 preparing a sample comprising serum or plasma.
2.2 transferring the sample into a centrifuge tube, and performing centrifugal operation, wherein the centrifugal speed is 3500-5000 rpm, and the centrifugal time is 7min.
2.3 taking supernatant to obtain serum or plasma samples.
S3: and detecting serum or plasma by using the kit, and obtaining the corresponding vancomycin drug concentration.
3.1 adding serum or plasma samples to be detected and reagents into the kit, and forming fluorescent signals after reaction.
3.2, detecting and quantitatively analyzing fluorescent signals by using instruments such as a chemiluminescent instrument and the like to obtain the corresponding concentration of the vancomycin drug.
As an optimization scheme of the embodiment of the invention, the method has the advantages that:
1. the operation is simple and convenient: the method is simple to operate, does not need complex instruments and techniques, and can rapidly obtain results.
2. The sensitivity is high: the chemical photoparticulate kit has high sensitivity and can rapidly and accurately detect the concentration of the vancomycin drug.
3. The accuracy is high: the method has accurate and reliable measurement result and high accuracy.
4. The operation cost is low: the kit has low preparation cost, is simple and convenient to operate, and can reduce detection cost.
As an optimization scheme of the embodiment of the invention, a new scheme for forming the parameter process is changed:
in step S2, the choice of centrifugation speed and time has an effect on the centrifugation effect of the sample, and a new protocol can be formed by adjusting the parameters of centrifugation and the formulation of the kit, as follows:
s2: taking 1-5 mL of sample to be detected, centrifuging at different centrifuging speeds and times, and taking supernatant to obtain serum or plasma.
2.1 preparing a sample comprising serum or plasma.
2.2 transferring the samples into centrifuge tubes, different centrifugation parameters were set, including centrifugation speed and time.
2.3 subjecting to centrifugation and collecting supernatant to obtain serum or plasma sample.
The scheme has the advantages that: through adjusting centrifugal parameters, impurities and precipitates in the sample can be removed better, and the purity of the sample is improved, so that the detection accuracy and sensitivity are improved.
To further optimize and refine the above scheme, the following can be considered:
1) Optimizing the formula of the kit: the detection sensitivity and accuracy can be further improved by optimizing the formulation of the kit. For example, the ratio of microparticles to other reagents in the kit can be adjusted, or new reagents can be added to improve detection performance.
2) Optimizing centrifugal parameters: by further optimizing the centrifugation parameters such as centrifugation time, centrifugation speed, etc., the purity and stability of the sample can be improved, thereby further improving the detection sensitivity and accuracy.
3) Repeatability and stability of the verification method: for the detection method, repeatability and stability among different laboratories, different operators and different batches of kits need to be verified so as to ensure the practical application effect of the method.
4) And (3) expanding a detection range: for different medicines, the formula and the detection method of the kit need to be optimized to meet the detection requirements of different medicines. Therefore, the range of vancomycin drug concentration detection needs to be expanded, and development and optimization of detection methods for other drugs are also needed.
5) The detection mechanism is studied in depth: in order to better understand the detection mechanism of the method, the reagent kit formula and the detection method can be optimized by deeply researching the chemical properties and the reaction mechanism of the reagent such as the microparticle reagent, the fluorescein and the like, so that the detection sensitivity and the detection accuracy are further improved.
6) Developing an automatic detection system: in order to improve detection efficiency and reduce human error, it is considered to develop an automated detection system, such as an automated fluorescence detector, for high throughput detection and automated data analysis.
7) Optimizing data analysis using artificial intelligence techniques: aiming at the analysis and processing problems of a large amount of data, the data can be analyzed and processed by utilizing artificial intelligence technology such as machine learning, deep learning and the like, so that the accuracy and the analysis efficiency of the data are improved. For example, drug concentration prediction models based on artificial intelligence techniques can be developed to help doctors and researchers understand the test results faster, thereby making more accurate diagnostic and therapeutic decisions.
In summary, the detection sensitivity, accuracy and reliability of the method can be further improved through optimization and refinement of the aspects of the reagent kit formulation, the centrifugal parameters, the detection mechanism, the data analysis and the like, so that the requirements of wider medicine quality control and clinical detection are met. Meanwhile, a new application field and a new technical innovation point can be explored, and new power is injected for the development of the field.
The foregoing is merely illustrative of specific embodiments of the present invention, and the scope of the invention is not limited thereto, but any modifications, equivalents, improvements and alternatives falling within the spirit and principles of the present invention will be apparent to those skilled in the art within the scope of the present invention.
Claims (10)
1. A method for determining vancomycin drug concentration by using a chemical photoparticulate reagent, the method comprising:
s1: preparing a chemical photoparticle kit;
s2: taking 1-5 mL of a sample to be detected, centrifuging for 7min at a centrifugation speed of 3500-5000 rpm, and taking supernatant to obtain serum or plasma;
s3: and detecting serum or plasma by using the kit, and obtaining the corresponding vancomycin drug concentration.
2. The method for determining vancomycin drug concentration by using the chemical light microparticle reagent according to claim 1, wherein the kit in S1 specifically comprises:
r1 reagent, R2 reagent, sample treatment fluid, magnetic separation reagent, cleaning fluid, eluent, calibrator and quality control product;
the reagent R1 is prepared from a biotinylated vancomycin monoclonal antibody A and a reagent diluent R1, the reagent R2 is prepared from an alkaline phosphatase-marked vancomycin monoclonal antibody B and a reagent diluent R2, the magnetic separation reagent is prepared from commercial streptavidin magnetic beads and a magnetic separation reagent diluent, and the calibrator and quality control materials are prepared from diluents containing different concentrations of vancomycin antigens and proteins; the chemiluminescent substrate solution needed in the experimental process is substrate solution for alkaline phosphatase catalytic luminescence.
3. The method for determining the concentration of the vancomycin drug by using the chemical photoparticulate reagent according to claim 2, wherein the working concentration of the biotinylated vancomycin monoclonal antibody A conjugate in the R1 reagent is 0.5-2.5 mug/mL; the pH value of the R1 reagent diluent is 7.5-8.0.
4. The method for determining vancomycin drug concentration using a chemical light microparticle reagent according to claim 2, wherein the components of the R1 reagent diluent comprise: tris, working concentration is 12.0-12.7 g/L; sodium azide with the working concentration of 0.48-0.6 g/L; sodium chloride with the working concentration of 5.7-6.0 g/L; bovine serum albumin with a working concentration of 9-10 g/L; methyl cellulose with working concentration of 0.7-1.0 g/L; the balance being purified water.
5. The method for determining the concentration of the vancomycin drug by using the chemical light microparticle reagent according to claim 2, wherein the working concentration of the alkaline phosphatase-labeled vancomycin monoclonal antibody B conjugate in the R2 reagent is 0.7-1.5 mug/mL.
6. The method for determining vancomycin drug concentration by using the chemical light microparticle reagent according to claim 2, wherein the pH value of the R2 reagent diluent is 7.7-8.0, and the method comprises the following steps: tris, working concentration is 12.0-14.3 g/L; sodium azide with the working concentration of 1.60-2.50 g/L; sodium chloride with the working concentration of 3.7-6.0 g/L; magnesium chloride with the working concentration of 0.5-0.5 g/L; zinc chloride with working concentration of 0.03-0.1 g/L; bovine serum albumin with a working concentration of 5-10 g/L; the working concentration of the AP enzyme protective agent is 400-500 g/L; the balance being purified water.
7. The method for determining vancomycin drug concentration by using the chemical light microparticle reagent according to claim 2, wherein the sample treatment solution is an aqueous solution of one or more than two of acetate buffer solution, acetonitrile and methanol, the content of ammonium acetate in the acetate buffer solution is 0.1-0.3 per mill, the content of methanol is 5-15%, and the content of acetonitrile is 5-15%.
8. The method for determining vancomycin drug concentration by using the chemical photoparticulate reagent of claim 2, wherein: the cleaning liquid is methanol or acetonitrile or aqueous solution of two mixtures;
the eluent is an aqueous solution of one or more than two of acetate buffer solution, acetonitrile and methanol, wherein the content of ammonium acetate in the acetate buffer solution is 0.05-0.15 per mill, the content of methanol is 0-15%, and the content of acetonitrile is 70-90%.
9. The method for determining vancomycin drug concentration by using the chemical light microparticle reagent according to claim 1, wherein the step S3 specifically comprises:
(1) Taking serum or plasma, transferring 1.0ml, adding into a sample bottle, and placing into an automatic sampler to sample 100ul for quantitative detection by liquid chromatography;
(2) Respectively adding an R1 reagent and an R2 reagent into a detection tube, and mixing;
(3) (37+ -0.5) deg.C for 15min;
(4) Adding 25 mu L of magnetic particle reagent into the detection tube;
(5) (37+ -0.5) deg.C for 5min;
(6) Adding 600 mu L of cleaning liquid into the detection tube, and uniformly mixing;
(7) Magnetically separating the supernatant to remove unreacted materials;
(8) Repeating the steps (6) - (7) twice;
(9) Adding 150 mu L of luminous substrate into a detection tube, and uniformly mixing for 3s;
(10) Measuring relative luminescence intensity (RLU) after ALP catalyzes luminescence of the substrate;
RLU and vancomycin antigen concentration are positively correlated in a certain range, and the vancomycin content of a sample to be detected is read from a standard curve by an interpolation method.
10. The method for determining the concentration of vancomycin drug using the chemical photoparticulate reagent of claim 1, comprising:
s1: reagent kit for preparing chemical photoparticulate
1.1 selecting suitable chemical reagents, including particulates and other reagents;
1.2 mixing the particles and other reagents according to a certain proportion to prepare; forming a kit;
s2: taking 1-5 mL of a sample to be detected, centrifuging for 7min at a centrifugation speed of 3500-5000 rpm, and taking supernatant to obtain serum or plasma;
2.1 preparing a sample comprising serum or plasma;
2.2, transferring the sample into a centrifuge tube, and performing centrifugal operation, wherein the centrifugal speed is 3500-5000 rpm, and the centrifugal time is 7min;
2.3 taking supernatant to obtain serum or plasma samples;
s3: detecting serum or plasma by using the kit, and obtaining the corresponding vancomycin drug concentration;
3.1, adding serum or plasma samples to be detected and a reagent into the kit, and forming a fluorescent signal after reaction;
3.2, detecting and quantitatively analyzing fluorescent signals by using instruments such as a chemiluminescent instrument and the like to obtain the corresponding concentration of the vancomycin drug.
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