CN113156136B - Kit for detecting immunoglobulin G in urine - Google Patents
Kit for detecting immunoglobulin G in urine Download PDFInfo
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- CN113156136B CN113156136B CN202110161710.2A CN202110161710A CN113156136B CN 113156136 B CN113156136 B CN 113156136B CN 202110161710 A CN202110161710 A CN 202110161710A CN 113156136 B CN113156136 B CN 113156136B
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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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
- G01N33/564—Immunoassay; Biospecific binding assay; Materials therefor for pre-existing immune complex or autoimmune disease, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/34—Genitourinary disorders
- G01N2800/347—Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
Abstract
The application discloses a kit for detecting immunoglobulin G in urine, belonging to the technical field of medical in-vitro diagnosis. The kit comprises a reagent R1 and a reagent R2; the reagent R1 comprises the following components: tris alkali, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, surfactant S9, EDTA-Na2And sodium benzoate; the reagent R2 comprises the following components: tris alkali, goat anti-human immunoglobulin G polyclonal antibody, animal serum, sodium benzoate, trehalose, EDTA-Na2And tween. The kit has high sensitivity, wide linear range and good repeatability, and can accurately detect low-value samples; meanwhile, the kit has better stability, the reagent can be stably stored for 14 months, the filtering step before the reagent is used is omitted, the operation steps are simplified, and the accuracy of the detection result is improved.
Description
Technical Field
The application relates to the technical field of medical in-vitro diagnosis, in particular to a kit for detecting immunoglobulin G in urine.
Background
Immunoglobulin (Ig) refers to a globulin having the activity or chemical structure of an antibody (Ab) similar to an antibody molecule. Immunoglobulin G is the major immunoglobulin in blood, mostly in the form of a monomer, mainly bound by spleen and lymph, and unable to be filtered through glomeruli, and thus is present in very low amounts in normal human urine.
IgG is higher in urine of patients with primary glomerulonephritis and chronic nephritis, and is only slightly increased in other types of glomerular diseases; IgM is only found in the urine of patients with chronic nephritis, but is found in very small amounts in the urine of patients with primary glomerulonephritis and latent nephritis. The type of Ig elevation in urine can be analyzed to help identify the type of glomerular disease being diagnosed.
In addition, the molecular weight of the IgG belongs to macromolecular protein of 150000 daltons, and if the excretion of the macromolecular protein in urine is increased, the heavier glomerulus injury is prompted, the permeability of a basement membrane is increased, so that the macromolecular IgG leaks out to become nonselective proteinuria, and the IgG is an important index of the abnormal selective permeability of a glomerular filtration membrane. Studies have suggested that urine IgG levels are positively correlated with urine protein, and urine IgG/blood IgG levels are positively correlated with proteinuria. With increasing risk levels of membranous nephropathy, urinary IgG also increases significantly, indicating more loss of mechanical barrier. Therefore, urine IgG level is also an index for the severity of membranous nephropathy.
At present, the methods for detecting the immunoglobulin G in the markets at home and abroad comprise ELISA and immunoturbidimetry. The principle of immunoturbidimetry is that IgG and corresponding antibody are specifically combined in a proper liquid phase to form a soluble antigen-antibody complex, and under the action of a polymerization promoter (such as polyethylene glycol and the like), the antigen-antibody complex is separated out from a solution to form particles, so that the reaction solution has turbidity. The immunoturbidimetry method can be used for directly analyzing samples in batches on a full-automatic biochemical analyzer due to the small sample consumption, is simple to operate and is widely applied clinically. However, the reagents and the construction method used by the existing immunoturbidimetry have some disadvantages and shortcomings, which are mainly shown in the following steps: in the process of preserving the antibody in the immunoturbidimetric reagent R2, flocculent precipitates are very easy to appear along with the time extension, the amount of the appearing flocculent precipitates is in direct proportion to the concentration of the antibody, and the flocculent precipitates are generally required to be removed by filtration, but the antibody can be removed after filtration, so that the performance of the antibody is poor, the linearity is insufficient, the repeatability of a detection sample is poor, and the Coefficient of Variation (CV) is increased, thereby causing the inaccurate detection result, delaying the illness state of a patient, increasing the filtration step, having complex operation and being incapable of meeting the requirements of clinical detection.
Disclosure of Invention
The application provides a kit for detecting immunoglobulin G in urine aiming at the problem that the antibody in the existing immunoturbidimetric reagent is unstable and easy to precipitate to influence the accuracy of a detection result.
The kit for detecting the immunoglobulin G in the urine is realized by the following technical scheme.
A kit for detecting immunoglobulin G in urine, comprising a reagent R1 and a reagent R2;
the reagent R1 comprises the following components:
the reagent R2 comprises the following components:
by adopting the technical scheme, the reagent kit is optimized and improved on the basis of components contained in a reagent R1 and a reagent R2 of the existing immunoturbidimetric kit, polyethylene glycol, polyvinylpyrrolidone and a surfactant S9 are added into the reagent R1 in a certain proportion, so that the sensitivity of the reagent is obviously improved, and meanwhile, the linear range of the reagent is widened, so that the linear range of the reagent kit is 0.2-80g/L, a sample with lower concentration can be detected, a sample with higher concentration can be directly detected on the premise of not needing to be diluted for many times, and the complexity of the detection process is reduced;
meanwhile, the reagent R2 is added with animal serum, sodium benzoate, trehalose and EDTA-Na according to a certain proportion2And Tween, the stability of the antibody is obviously improved, and the reagent is not turbid in the long-term low-temperature storage process, so that the pretreatment step of filtering the reagent before use is omitted. The reagent kit can effectively ensure the stability of the reagent, the reagent can be stably stored for 14 months without influencing the performance of the reagent, the reagent stored for 14 months is adopted to detect a sample to be detected, and the detection result still has higher accuracy.
In summary, the kit has high sensitivity, wide linear range and excellent repeatability, can detect samples with low concentration, and avoids the occurrence of false negative detection results and the delay of the patient's condition; on the other hand, the kit has better stability, the detection reagent can be stored for 14 months, and the detection result is still accurate and reliable.
Optionally, the reagent R1 comprises the following components:
by adopting the technical scheme, the content of each component in the reagent R1 is further optimized, the analysis sensitivity of the kit is further improved, and the accuracy of the reagent in a linear range of 0.2-80g/L is further improved.
Optionally, the reagent R2 comprises the following components:
through adopting above-mentioned technical scheme, the content of each component in reagent R2 is further optimized in this application, and the stability of kit further improves, and reagent R2 further reduces depositing the incidence that the in-process appears the flocculent deposit, and the turbidity of reagent further reduces, has avoided filtering the influence to the reagent performance, has improved the sensitivity of kit from on the other hand, has improved the degree of accuracy that low value sample detected, and the repeatability is better, fine control coefficient of variation.
Optionally, the pH value of the reagent R1 and the reagent R2 is 7.2-7.6.
By adopting the technical scheme, the pH values of the reagent R1 and the reagent R2 are controlled to be 7.2-7.6, so that a stable reaction environment and a stable storage environment can be provided for the antigen and the antibody, and the accuracy of a detection result is improved.
Optionally, the animal serum is any one of horse serum, bovine serum and chicken serum.
Through adopting above-mentioned technical scheme, the animal serum of this application chooses horse serum, ox serum and chicken serum for use, and these three kinds of sera add in reagent R2, can simulate the internal environment of organism, provide more suitable preservation environment for immunoglobulin G polyclonal antibody, have improved the stability of antibody, have prolonged the shelf life of antibody, make reagent R2 be difficult to appear muddy phenomenon.
Optionally, the polyethylene glycol is selected from one or more of polyethylene glycol 2000, polyethylene glycol 4000, polyethylene glycol 6000 and polyethylene glycol 8000.
By adopting the technical scheme, the polyethylene glycol with different molecular weights can be selected for use and added into the reagent R1, so that other components in the reagent R1 can be fully dispersed, and the reagent R1 has good uniformity.
Optionally, the tween is one or two of tween 20 and tween 80.
Through adopting above-mentioned technical scheme, tween 20 and tween 80 are selected for use to the tween of this application, add it into reagent R2, can make the abundant dispersion of other each component in the reagent R2, have improved reagent R2's degree of consistency, can provide stable environment for the antibody, prevent the emergence of sediment.
Optionally, the kit further comprises a calibrator and a quality control, wherein the calibrator and the quality control are samples containing immunoglobulin G antigens with different concentrations.
By adopting the technical scheme, the kit further comprises a calibrator and a quality control product, the calibrator is used for drawing a calibration curve, the concentration of the sample to be measured can be obtained by substituting the absorbance change value obtained by measurement into the calibration curve, and the measurement method is simple; the quality control product is arranged to indicate the effectiveness of the reagents R1 and R2 in the kit, so that the accuracy and reliability of the detection result are improved.
Optionally, in the calibrator and the quality control product, the immunoglobulin G antigen diluent comprises the following components: PBS buffer solution with final concentration of 20mmol/L, animal serum with final concentration of 10-50ml/L, sodium benzoate with final concentration of 5-10g/L, Tween with final concentration of 0.5-1ml/L, and pH of the solution is 7.2.
By adopting the technical scheme, the components of the immunoglobulin G antigen diluent in the calibrator and the quality control product are further optimized, the optimized diluent can dilute the immunoglobulin G antigen to a proper working concentration, and can provide a stable working environment for the immunoglobulin G antigen, so that the immunoglobulin G antigen can be better combined with an immunoglobulin G polyclonal antibody, and the immunoglobulin G antigen diluent is used for drawing a calibration curve and detecting the effectiveness of a reagent.
Optionally, the volume ratio of the sample to be detected to the reagents R1 and R2 in the reaction system is 3:240: 60; the main wavelength is 600nm and the secondary wavelength is 800 nm.
By adopting the technical scheme, the reaction system and the detection condition are further optimized, and the sample is detected under the reaction system and the detection condition, so that the detection sensitivity and specificity are higher, and the method is more suitable for detecting the sample with lower antigen content.
In summary, the present application has the following beneficial effects:
1. by optimizing and improving the components of the reagent R1, the sensitivity of the reagent is obviously improved, the detection limit is 0.2g/L, and the linear range of the reagent is widened, wherein the linear range is 0.2-80g/L, so that the kit can be used for detecting low-value samples, high-value samples can be directly detected without being diluted for many times, and the operation steps are simpler;
2. this application is optimized and is improved through the composition to reagent R2, has obviously improved antibody stability, makes reagent a large amount of muddy can not appear in long-term save process, and reagent can stably be preserved 14 months, has saved the filtering step of reagent before using, has improved the accuracy of testing result when having simplified the testing step.
Drawings
FIG. 1 is a graph of the linear correlation of the kit of example 1 of the present application;
FIG. 2 is a graph of the linear dependence of the kit of example 2 of the present application.
Detailed Description
The present application will be described in further detail with reference to the following drawings and examples.
The polyclonal antibody of goat anti-human immunoglobulin G is purchased from Beijing Youthful Biotechnology GmbH;
the immunoglobulin G antigen of the present application was purchased from beijing apiss biotechnology limited;
the model of the full-automatic biochemical analyzer is Hitachi 7170;
surfactant S9(TETRONIC 1307) of the present application was purchased from Sigma-Aldrich;
horse serum, chicken serum and bovine serum of the present application were purchased from Sigma-Aldrich;
immunoturbidimetric control reagents of the present application were purchased from beijing baiolabockik technologies ltd.
Example 1
A kit for detecting immunoglobulin G in urine, comprising a reagent R1 and a reagent R2;
the preparation method of the reagent R1 comprises the following steps: weighing 2.42g of Tris alkali, 600040 g of polyethylene glycol, 30g of polyvinylpyrrolidone, 9g of sodium chloride, S91 g serving as surfactant and EDTA-Na210g and 5g of sodium benzoate, adding water to 950mL, adjusting the pH value of the solution to 7.6, and metering the volume to 1L, namely obtaining a reagent R1.
The preparation method of the reagent R2 comprises the following steps: weighing Tris alkali 2.42G, goat anti-human immunoglobulin G polyclonal antibody 100ml, horse serum 10ml, sodium benzoate 5G, trehalose 5G, EDTA-Na210g and 200.5 mL of Tween, adding water to 950mL, adjusting the pH value of the solution to 7.6, and metering the volume to 1L, namely the reagent R2.
The preparation method of the immunoglobulin G antigen diluent comprises the following steps: 200mL of 10 gamma PBS solution, 50mL of horse serum, 5g of sodium benzoate and 1mL of Tween are taken, water is added to 950mL, the pH value of the solution is adjusted to 7.2, and finally the volume is fixed to 1L.
A calibrator dilution method:
the immunoglobulin G antigen was diluted to 80G/L, 40G/L, 20G/L, 10G/L, 5G/L, and 0G/L, respectively, with a diluent, for a total of 6 levels.
The quality control product dilution method comprises the following steps:
the immunoglobulin G antigen was diluted to 6G/L and 12G/L, respectively, at 2 levels.
Adding 1240 microliter of reagent R, 260 microliter of reagent R and 3 microliter of sample to be detected into the reaction system, adopting a full-automatic biochemical analyzer to measure the absorbance of the sample to be detected, measuring the main wavelength of 600nm and the auxiliary wavelength of 800nm, reading the readings of 16-34 points by a two-point end point method and calculating.
1. Drawing of calibration curve
And (3) carrying out absorbance detection on the calibrator in a full-automatic biochemical analyzer, wherein specific detection data are shown in table 1.
TABLE 1 calibration Curve results
| Level of | S1 | S2 | S3 | S4 | S5 | S6 |
| Concentration (g/L) | 0 | 5 | 10 | 20 | 40 | 80 |
| Value of absorbance change | 41 | 1600 | 2907 | 4796 | 5607 | 6105 |
2. Linearity detection
Linear high-value samples are selected and diluted into six levels of 80g/L, 40g/L, 20g/L, 5g/L, 2.5g/L and 0.2g/L by low-value samples; three tests were performed per sample using a fully automated biochemical analyzer and the relative deviation and correlation coefficient were calculated, for specific experimental results see table 2. With the theoretical concentration as the abscissa and the average value of the detected concentration as the ordinate, a linear correlation chart is drawn, see fig. 1.
TABLE 2 Linear Range dilution
As can be seen from Table 2 and FIG. 1, the kit of the present embodiment is used for detecting the sample, the obtained detection concentration is very close to the theoretical concentration, the linearity is very good, the linearity range is between 0.2 and 80g/L, the correlation coefficient R is not less than 0.990, and the detection result is accurate and reliable.
3. Repeatability detection
And (3) carrying out absorbance detection on the quality control substances of two concentration levels by using a full-automatic biochemical analyzer, detecting each concentration level for 10 times, and calculating the standard deviation and the variation coefficient, wherein specific experimental results are shown in a table 3.
TABLE 3 measurement repeatability
| Theoretical value | 6g/L | 12g/L |
| 1 time of | 6.02 | 11.83 |
| 2 times (one time) | 6.05 | 11.99 |
| 3 times of | 6.04 | 11.75 |
| 4 times (twice) | 6.03 | 11.85 |
| 5 times (twice) | 6.10 | 11.95 |
| 6 times of | 6.01 | 11.78 |
| 7 times (twice) | 6.06 | 11.85 |
| 8 times (by volume) | 6.13 | 11.99 |
| 9 times of | 6.05 | 11.95 |
| 10 times of | 6.04 | 11.76 |
| Mean value | 6.05 | 11.87 |
| SD | 0.03 | 0.09 |
| CV％ | 0.57 | 0.75 |
As can be seen from Table 3, the kit of the present embodiment is used for repeated detection of 6g/L and 12g/L samples, and the coefficient of variation CV of the obtained result is less than 5%, so that the kit meets the requirements, has excellent repeatability, and has high accuracy of the detection result.
4. Stability detection
Reagents R1, R2 and calibrators of this example were stored at 2-8 ℃ and were calibrated every three months and the absorbance change was recorded for each concentration of calibrators and reagent blanks and the results are shown in table 4.
Table 4 long term stability data results
It can be seen from table 4 that the reagent R1, the reagent R2 and the calibrator of the kit are stored for 420 days at 2-8 ℃, the absorbance change of each horizontal calibrator and the blank reagent is within 10% of the absorbance change of the initial preparation, no obvious flocculent precipitate appears in the reagent R2, the reagent has good stability, the reagent can be directly used without filtration after long-term storage, the antibody performance is not affected, the linearity and the repeatability of the detection result are kept at high levels, and the use validity of the kit is long.
5. Sensitivity test
The purpose of the sensitivity experiment is to detect the absorbance change value of the reagent of the kit when testing physiological saline and a certain concentration of clinical samples.
The kit and control reagent (immunoturbidimetry) of this example were used to determine the absorbance change of 0.9% saline and 5g/L clinical samples (urine samples), each sample was tested 3 times, and the experimental results are shown in Table 5.
TABLE 5 comparison of reagent sensitivity
As can be seen from Table 5, the change in absorbance of the saline solution measured by using the kit of this example was 39 (10000A); when a urine sample with a theoretical concentration of 5g/L was measured, the absorbance change value was 1620 (10000A). Measuring the physiological saline by using a contrast reagent, wherein the absorbance change value is 91 (10000A); when a urine sample with a theoretical concentration of 5g/L was measured, the absorbance change value was 572 (10000A). The kit has the advantages that the difference between the absorbance change value of the clinical sample and the blank control signal value is large, the detection result of a low-value part is more sensitive, the contrast reagent is adopted to detect that the difference between the absorbance change value of the clinical sample and the blank control signal value is small or close, and the value of 0 is easily detected for the low-value part. Therefore, the sensitivity of the kit is obviously higher than that of the common immunoturbidimetric reagent.
Example 2
A kit for detecting immunoglobulin G in urine, comprising a reagent R1 and a reagent R2;
the preparation method of the reagent R1 comprises the following steps: weighing 6.057g of Tris alkali, 400050 g of polyethylene glycol, 40g of polyvinylpyrrolidone, 9g of sodium chloride, S92 g as a surfactant and EDTA-Na220g and 10g of sodium benzoate, adding water to 950mL, adjusting the pH value of the solution to 7.2, and metering the volume to 1L, namely obtaining a reagent R1.
The preparation method of the reagent R2 comprises the following steps: weighing 6.057g Tris base and goat anti-human immunity200ml of globulin G polyclonal antibody, 50ml of bovine serum, 10G of sodium benzoate, 3G of trehalose and EDTA-Na220g and Tween 801 mL, adding water to 950mL, adjusting the pH of the solution to 7.2, and metering the volume to 1L to obtain a reagent R2.
The preparation method of the immunoglobulin G antigen diluent comprises the following steps: 200mL of PBS solution with 10 gamma, 10mL of bovine serum, 10g of sodium benzoate and 0.5mL of Tween are taken, water is added to 950mL, the pH value of the solution is adjusted to 7.2, and finally the volume is fixed to 1L.
A calibrator dilution method:
the immunoglobulin G antigen was diluted to 80G/L, 40G/L, 20G/L, 10G/L, 5G/L, and 0G/L, respectively, with a diluent, for a total of 6 levels.
The quality control product dilution method comprises the following steps:
the immunoglobulin G antigen was diluted to 6G/L and 12G/L, respectively, at 2 levels.
Adding 1240 microliter of reagent R, 260 microliter of reagent R and 3 microliter of sample to be detected into the reaction system, adopting a full-automatic biochemical analyzer to measure the absorbance of the sample to be detected, measuring the main wavelength of 600nm and the auxiliary wavelength of 800nm, and reading two points of 16-34 by a two-point end point method for calculation.
1. Drawing of calibration curve
And (3) carrying out absorbance detection on the calibrator in a full-automatic biochemical analyzer, wherein specific detection data are shown in table 1.
TABLE 6 calibration Curve results
| Level of | S1 | S2 | S3 | S4 | S5 | S6 |
| Concentration (g/L) | 0 | 5 | 10 | 20 | 40 | 80 |
| Value of absorbance change | 41 | 2650 | 3921 | 4986 | 6087 | 8034 |
2. Linearity detection
Linear high-value samples are selected and diluted into six levels of 80g/L, 60g/L, 30g/L, 10g/L, 2g/L and 0.2g/L by low-value samples; three tests were performed per sample using a fully automated biochemical analyzer and the relative deviation and correlation coefficient were calculated, for specific experimental results see table 7. With the theoretical concentration as the abscissa and the average value of the detected concentration as the ordinate, a linear correlation chart is drawn, see fig. 2.
TABLE 7 Linear Range dilution
As can be seen from Table 7 and FIG. 2, when the kit of the present embodiment is used for detecting a sample to be detected, the obtained detection concentration is very close to the theoretical concentration, the linearity is very good, the linearity range is between 0.2 and 80g/L, the correlation coefficient R is not less than 0.990, the linearity requirement is satisfied, and the detection result is accurate and reliable.
3. Repeatability detection
And (3) carrying out absorbance detection on the quality control substances of two concentration levels by using a full-automatic biochemical analyzer, detecting each concentration level for 10 times, and calculating the standard deviation and the variation coefficient, wherein specific experimental results are shown in a table 8.
TABLE 8 measurement repeatability
| Theoretical value | 6g/L | 12g/L |
| 1 time of | 6.12 | 12.02 |
| 2 times (one time) | 6.13 | 12.12 |
| 3 times of | 6.08 | 12.03 |
| 4 times (twice) | 6.13 | 12.09 |
| 5 times (twice) | 6.09 | 12.03 |
| 6 times of | 6.01 | 12.12 |
| 7 times (twice) | 6.16 | 12.13 |
| 8 times (by volume) | 6.03 | 12.09 |
| 9 times of | 6.15 | 12.08 |
| 10 times of | 6.14 | 11.98 |
| Mean value | 6.10 | 12.07 |
| SD | 0.05 | 0.05 |
| CV％ | 0.79 | 0.40 |
As can be seen from Table 8, the kit of the present embodiment is used for repeated detection of 6g/L and 12g/L samples, and the coefficient of variation CV of the obtained result is less than 5%, so that the kit meets the requirements, has excellent repeatability and higher accuracy of the detection result.
4. Detection limit determination
The kit of the embodiment is used for detecting 5 low-value samples with the concentration of 0.2-0.3g/L, each sample is detected for 5 times, the detection results are counted, the number of the detection results which are lower than the blank limit of 0.08g/L is less than or equal to 3, no detection result which is higher than 0.5g/L exists, and the specific statistical results are shown in Table 9.
TABLE 9 detection Limit results
| 1 time of | 2 times (one time) | 3 times of | 4 times (twice) | 5 times (twice) | |
| Sample 1 | 0.21 | 0.2 | 0.19 | 0.18 | 0.19 |
| Sample 2 | 0.21 | 0.18 | 0.17 | 0.16 | 0.17 |
| Sample 3 | 0.19 | 0.21 | 0.2 | 0.18 | 0.17 |
| Sample 4 | 0.21 | 0.2 | 0.19 | 0.18 | 0.16 |
| Sample 5 | 0.05 | 0.2 | 0.18 | 0.17 | 0.16 |
As can be seen from Table 9, when the kit of the present embodiment is used to detect 5 low-value samples, each sample is detected 5 times, and in the detection results, the number of the detection results that are lower than the blank limit by 0.08g/L is 0, which meets the requirement that the number of the detection results is less than or equal to 3, and at the same time, no detection result that is higher than 0.5g/L exists, and the detection results meet the requirement. Therefore, the detection limit of the kit is not higher than 0.2g/L, the detection sensitivity is high, even when the antigen content in the sample to be detected is low, the antigen-antibody conjugate is less, and the turbidity degree is low, the kit can still detect the content of IgG in the sample to be detected, and can effectively avoid false negative of the detection result, thereby delaying the occurrence of the state of illness of the patient.
The embodiments of the present invention are preferred embodiments of the present application, and the scope of protection of the present application is not limited by the embodiments, so: all equivalent changes made according to the structure, shape and principle of the present application shall be covered by the protection scope of the present application.
Claims (5)
1. A kit for detecting immunoglobulin G in urine, which is characterized in that: comprises a reagent R1 and a reagent R2;
the reagent R1 comprises the following components:
tris alkali 2.42 g/L;
polyethylene glycol 600040 g/L;
30g/L of polyvinylpyrrolidone;
9 g/L of sodium chloride;
surfactant S91 g/L;
EDTA-Na2 10 g/L；
5g/L of sodium benzoate;
the reagent R2 comprises the following components:
tris alkali 2.42 g/L;
100ml/L of goat anti-human immunoglobulin G polyclonal antibody;
10 ml/L horse serum;
5g/L of sodium benzoate;
5g/L of trehalose;
EDTA-Na2 10 g/L；
tween 200.5 ml/L.
2. The kit for detecting immunoglobulin G in urine according to claim 1, wherein: the pH value of the reagent R1 and the reagent R2 is 7.2-7.6.
3. The kit for detecting IgG in urine according to any one of claims 1-2, wherein: the kit also comprises a calibrator and a quality control product, wherein the calibrator and the quality control product are samples containing immunoglobulin G antigens with different concentrations.
4. The kit for detecting IgG in urine according to claim 3, wherein: in the calibrator and the quality control product, the immunoglobulin G antigen diluent comprises the following components: PBS buffer solution with final concentration of 20mmol/L, animal serum with final concentration of 10-50ml/L, sodium benzoate with final concentration of 5-10g/L, Tween with final concentration of 0.5-1ml/L, and pH of the solution is 7.2.
5. The kit for detecting IgG in urine according to any one of claims 1-2, wherein: the volume ratio of the sample to be detected to the reagent R1 and the reagent R2 in the reaction system is 3:240: 60; the main wavelength is 600nm and the secondary wavelength is 800 nm.
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|---|---|---|---|---|
| CN104777319A (en) * | 2015-05-04 | 2015-07-15 | 山东博科生物产业有限公司 | Immune globulin G immunoturbidimetry detection kit |
| CN107015004A (en) * | 2017-03-21 | 2017-08-04 | 深圳市汇松科技发展有限公司 | A kind of serum amyloid A protein determines kit and preparation method thereof |
| CN108845147A (en) * | 2018-07-04 | 2018-11-20 | 浙江伊利康生物技术有限公司 | A kind of cystatin C detection kit |
| CN109633172A (en) * | 2019-01-03 | 2019-04-16 | 迪瑞医疗科技股份有限公司 | Human urine immunoglobulin G detection reagent box based on Immunoturbidimetry |
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2021
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