CN113740539B - Kit for determining specific growth factors - Google Patents
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Abstract
The invention discloses a kit for determining a specific growth factor, which comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises a buffer solution, a surfactant 1, a surfactant 2, inorganic salts and an organic solvent, and the reagent R2 comprises a buffer solution, a color developing agent and a preservative. The invention can effectively promote the forward progress of the ninhydrin color reaction by adding a certain amount of organic solvent (DMSO) into the detection reagent, so that the color reaction is close to complete reaction. Meanwhile, the problem of subsequent sample precipitation caused by adding the organic solvent is solved by adding the composition of the surfactant and the salt. The detection performance of the kit is effectively improved by matching the improved measures with the reaction system, wherein the detection performance comprises the precision and the quantifiability of the object to be detected in a linear range.
Description
Technical Field
The invention relates to the field of medical examination, in particular to a kit for measuring specific growth factors.
Background
Specific growth factors (Specific GrowthFanctor, SGF) are a special class of substances, including saccharides, amino acids and metabolites thereof, and have a characteristic that when a tumor is produced, the content of the Specific growth factors in blood is increased, so that the Specific growth factors can stimulate the generation of malignant tumor blood vessels, and after the generation of the blood vessels, enough nutrition can be provided for tumor tissues, so that the rapid growth of the tumor is promoted. The specific growth factor can be obviously increased in the early stage of the tumor, and the content of the specific growth factor in serum is clinically measured to be used as an effective reference basis for early detection of the tumor, change of the patient's condition and postoperative monitoring.
Currently known SGF detection means include an enzyme-linked immunosorbent assay (ELISA) and a common chemical method, wherein the ELISA has the disadvantages of complex operation, long reaction time and poor detection repeatability, so that the ELISA is not favorable for wide clinical application. The common chemical method utilizes the mixed solution in the kit to carry out superposition coupling reaction with specific growth factors (specific glycolipid with amino groups, metabolic products of glycoprotein, free alpha-amino acid and other substances) in a sample to be detected at the temperature of 37 ℃, the reaction product and a chromogenic agent in the kit carry out chromogenic reaction under the same reaction condition to form a blue product, the absorbance value of the chromogenic product is measured at the wavelength of 570m, and the content of the specific growth factors in the sample to be detected is calculated through a standard concentration curve and a formula. Since the color reaction of the color reagent ninhydrin in the reagent is a reversible reaction and is incomplete, the problem of poor metering property and thus linear disqualification of the detection reagent used clinically at present exists generally.
Disclosure of Invention
The inventors have unexpectedly found that by adding a certain amount of an organic solvent (DMSO) to the detection reagent, the forward progress of the ninhydrin color reaction can be effectively promoted, so that the color reaction is near complete. Meanwhile, the problem of subsequent sample precipitation caused by adding the organic solvent is solved by adding the composition of the surfactant and the salt. The detection performance of the kit is effectively improved by matching the improved measures with the reaction system, wherein the detection performance comprises the precision and the quantifiability of the object to be detected in a linear range.
In order to achieve the above purpose, the present invention adopts the following technical means: a kit for determining a specific growth factor, comprising reagent R1 and reagent R2, characterized in that: the reagent R1 comprises a buffer solution, a surfactant 1, inorganic salts and an organic solvent.
Preferably, the reagent R2 comprises a buffer solution, a color developing agent and a preservative.
Preferably, the organic solvent is selected from at least one of DMSO, DMF or methanol, preferably DMSO.
Preferably, the content of the organic solvent is 200-600g/L, preferably 400-600g/L.
Preferably, the reagent R1 also comprises a surfactant 2; the reagent R2 comprises a buffer solution, a color developing agent and a preservative.
Preferably, the surfactant 1 and the surfactant 2 are selected from two of Triton X-100, triton X-305, emulgen LS-114, emulgen 709, tween-20 or Tween-80, preferably Triton X-100 and Emulgen LS-114, triton X-305 and Emulgen LS-114 or a combination of Triton X-305 and Tween-20; the content of the surfactant 1 is 5-20g/L, and the content of the surfactant 2 is 0.1-1g/L.
Preferably, the buffer solution in the reagent R1 is at least one selected from 4-hydroxyethyl piperazine ethane sulfonic acid, tris buffer solution or GOOD' S buffer solution, and the inorganic salt is at least one selected from sodium chloride or potassium chloride; the buffer solution in the reagent R2 is at least one of phosphate buffer solution, GOOD' S buffer solution or succinic acid buffer solution, the color-developing agent is ninhydrin, and the preservative is at least one of sodium azide or Proclin 300; the content of the buffer solution in the reagent R1 is 50-150mM, and the content of the inorganic salt is 1-5g/L; the content of the buffer solution in the reagent R2 is 30-50mM, the content of the color developing agent is 10-100mM, and the content of the preservative is 0.1-1g/L.
A composition effective to enhance the detection of biochemical agents, said composition comprising: an organic solvent, a surfactant 1, a surfactant 2 and an inorganic salt.
Preferably, the organic solvent is selected from at least one of DMSO, DMF or methanol, preferably DMSO; the surfactant 1 and the surfactant 2 are selected from two of Triton X-100, triton X-305, emulgen LS-114, emulgen 709, tween-20 or Tween-80, preferably Triton X-100 and Emulgen LS-114, triton X-305 and Emulgen LS-114 or a combination of Triton X-305 and Tween-20; the inorganic salt is at least one selected from sodium chloride and potassium chloride.
Preferably, the reagent is a kit for detecting a specific growth factor.
The invention has the beneficial effects that: according to the invention, a certain amount of organic solvent (DMSO) is added into the detection reagent R1, so that forward progress of the ninhydrin color reaction is effectively promoted, the color reaction is close to complete reaction, and the quantifiability of the detection reagent to the substance to be detected in a linear range is remarkably improved. Meanwhile, the problems of sample precipitation and detection precision reduction are brought along with the addition of the organic solvent, and further the problems of subsequent sample precipitation and precision reduction after the addition of the organic solvent are solved by adding the composition of the surfactant and the salt in the reagent R1, so that the detection performance of the kit is effectively improved.
Drawings
FIG. 1 is a graph showing the linear relationship between the theoretical concentration and the detection value of the sample in the group A kit in example 3 of the present invention;
FIG. 2 is a graph showing the linear relationship between the theoretical concentration and the detection value of the sample in the B-group kit according to the embodiment 3 of the present invention;
FIG. 3 is a graph showing the linear relationship between the theoretical concentration and the detection value of the sample in the group C kit according to the embodiment 3 of the present invention;
FIG. 4 is a graph showing the linear relationship between the theoretical concentration and the detection value of the sample in the group A kit in example 4 of the present invention;
FIG. 5 is a graph showing the linear relationship between the theoretical concentration and the detection value of the B group kit sample in example 4 of the present invention;
FIG. 6 is a graph showing the linear relationship between the theoretical concentration and the detection value of the sample in the group C kit according to the embodiment 4 of the present invention;
FIG. 7 is a graph showing the relationship between the theoretical concentration of the samples of the group D kit and the detection value in example 4 of the present invention;
FIG. 8 is a graph showing the linear relationship between the theoretical concentration and the detection value of the E group kit sample in example 4 of the present invention;
FIG. 9 is a graph showing the linear relationship between the theoretical concentration and the detection value of the F group kit sample in example 4 of the present invention;
FIG. 10 is a graph showing the linear relationship between the theoretical concentration and the detection value of the G group kit sample in example 4 of the present invention;
FIG. 11 is a graph showing the linear relationship between the theoretical concentration and the detection value of the H-group kit sample in example 4 of the present invention;
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
The following examples are included to more clearly and clearly illustrate the technical aspects of the present invention by way of illustration. Those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. The detailed description of the invention is merely illustrative of the invention and is not intended to be limiting in any way.
Example 1 preparation of TSGF detection kit
The Specific Growth Factor (TSGF) detection kit of the present invention includes two liquid components of reagent R1 and reagent R2 that are independent of each other.
1. Preparation of reagent R1
The preparation method comprises the steps of preparing the composition according to the following formula, fully stirring and uniformly mixing, and storing at 2-8 ℃.
2. Preparation of reagent R2
The preparation method comprises the steps of preparing the composition according to the following formula, fully stirring and uniformly mixing, and storing at 2-8 ℃.
Example 2 methods of Using the kit
In this example, a full-automatic biochemical analyzer (Hitachi 7180) was used in combination with the kit of the present invention for sample detection.
(1) Instrument parameter setting
(2) Assay protocol
(3) Calculation method
Using a two-point linear calibration mode, taking a linear function as a calculation mode, and making a dose/response curve according to the value of the calibrator and the absorbance change value, wherein the content of the specific growth factor in the sample can be calculated on the dose/response curve according to the absorbance change value.
The detection principle of the invention is as follows: the mixed solution in the kit is subjected to superposition coupling reaction with specific growth factors (specific glycolipid with amino groups, glycoprotein metabolite, free alpha-amino acid and other substances) in a sample to be detected at the temperature of 37 ℃, the reaction product and a chromogenic agent in the kit are subjected to chromogenic reaction under the same reaction condition to form a blue product, the absorbance value of the chromogenic product is measured at the wavelength of 570nm, and the content of the specific growth factors in the sample to be detected is converted through a standard concentration curve and a formula (TSGF clinical concentration=100-0.18 TSGF equivalent concentration).
Example 3 Performance test of kit
In order to verify each performance of the kit, 3 groups of kits are arranged for performance verification:
group A: the kit prepared in the embodiment 1 of the invention;
group B: specific growth factor assay kit (chemical method) (beijing jiugang);
group C: a kit obtained according to the method described in example 1 of the description of patent CN 104515768B.
The kit of group C adopts the existing reagents of the laboratory (namely, the same components are involved in the kit of group A and the kit of group C, for example, the ninhydrin as a color reagent is produced by the same manufacturer and in the same batch number), wherein the kit of group A is tested according to the use method described in example 2, the kit of group B is tested according to the instruction of the kit of group B, and the kit of group C is tested according to the use method in the example of the kit of group C.
(1) Accuracy verification
And (3) respectively carrying out accuracy test on clinical assignment samples by using three groups of kits, setting 2 repetitions, reading signals by a full-automatic biochemical analyzer (Hitachi 7180), and calculating the relative deviation between the measured average value and the target value to carry out accuracy verification. The test results are shown in the following table:
table 1 accuracy verification
From the above experimental results, the relative deviations of the three sets of kit test values 1 and target value 1 were respectively 0.21%, 0.61% and-2.12%, and the relative deviations of the test value 2 and target value 2 were respectively-0.25%, 0.71% and-1.13%. The detection accuracy of the kit (A group) prepared in the embodiment 1 of the invention is obviously better than that of the kit (C group).
(2) Precision verification
And (3) selecting a low-value sample, a median sample and a high-value sample of the clinical specific growth factor, respectively testing the samples by using three groups of kits, respectively repeatedly measuring each sample for 10 times, reading signals by a full-automatic biochemical analyzer (Hitachi 7180), respectively calculating a measurement mean value and a standard deviation, and calculating a variation coefficient to carry out precision investigation. The test results are shown in the following table:
table 2 precision verification
From the above experimental results, the three groups of kits have a variation coefficient of 0.59%, 2.87% and 2.30% when detecting low-value samples, a variation coefficient of 0.49%, 0.53% and 1.91% when detecting median samples, and a variation coefficient of 0.58%, 1.00% and 2.87% when detecting high-value samples, respectively, and the experimental results show that the kit (group a) prepared in the embodiment 1 of the present invention has a precision superior to that of the control kit-1 (group B) and the control kit-2 (group C) when detecting low-value samples, median samples and high-value samples.
(3) Linear range verification
And selecting a clinical ultrahigh value sample and a low value sample, then configuring each concentration gradient sample proportionally by using the high value sample and the low value sample, respectively testing the samples by using three groups of kits, respectively and repeatedly measuring each sample for 2 times, reading signals by a full-automatic biochemical analyzer (Hitachi 7180), and respectively calculating measurement mean values to perform linear range investigation. The test results are shown in the following table:
table 3A set of kit Linear Range verification
| H-L | L-H | Theoretical value | Mean value of | Relative deviation 1 | Estimated value | Relative deviation 2 | |
| H | 283.94 | 282.72 | 283.33 | 283.33 | 0.00％ | 282.81 | 0.18％ |
| 3/4H+1/4L | 241.81 | 237.19 | 241.20 | 239.50 | -0.71％ | 240.58 | -0.45％ |
| 1/2H+1/2L | 198.5 | 201.08 | 199.08 | 199.79 | 0.36％ | 198.35 | 0.72％ |
| 1/4H+3/4L | 157.41 | 152.02 | 156.95 | 154.72 | -1.42％ | 156.12 | -0.90％ |
| 1/8H+7/8L | 133.9 | 134.61 | 135.89 | 134.26 | -1.20％ | 135.01 | -0.56％ |
| 1/16H+15/16L | 125.73 | 123.87 | 125.36 | 124.80 | -0.44％ | 124.45 | 0.28％ |
| L | 114.08 | 115.57 | 114.83 | 114.83 | 0.00％ | 113.89 |
Note that: the relative deviation 1 is the relative deviation of the mean value from the theoretical value, and the relative deviation 2 is the relative deviation of the mean value from the estimated value.
Table 4B set of kit Linear Range verification
| H-L | L-H | Theoretical value | Mean value of | Relative deviation 1 | Estimated value | Relative deviation 2 | |
| H | 295.25 | 301.8 | 298.53 | 298.53 | 0.00％ | 287.32 | 3.90％ |
| 3/4H+1/4L | 220.15 | 231.96 | 240.60 | 226.06 | -6.04％ | 229.64 | -1.56％ |
| 1/2H+1/2L | 161.7 | 154.23 | 182.67 | 157.97 | -13.52％ | 171.95 | -8.14％ |
| 1/4H+3/4L | 107.54 | 106.52 | 124.74 | 107.03 | -14.20％ | 114.27 | -6.34％ |
| 1/8H+7/8L | 86.67 | 86.77 | 95.77 | 86.72 | -9.45％ | 85.43 | 1.51％ |
| 1/16H+15/16L | 72.61 | 73.60 | 81.29 | 73.11 | -10.07％ | 71.01 | 2.95％ |
| L | 65.87 | 67.75 | 66.81 | 66.81 | 0.00％ | 56.59 |
Note that: the relative deviation 1 is the relative deviation of the mean value from the theoretical value, and the relative deviation 2 is the relative deviation of the mean value from the estimated value.
Table 5C set of kit Linear Range verification
| H-L | L-H | Theoretical value | Mean value of | Relative deviation 1 | Estimated value | Relative deviation 2 | |
| H | 247.56 | 248.15 | 247.86 | 247.86 | 0.00％ | 248.92 | -0.43％ |
| 3/4H+1/4L | 210.36 | 204.58 | 214.60 | 207.47 | -3.32％ | 216.04 | -3.97％ |
| 1/2H+1/2L | 195.5 | 201.08 | 181.34 | 198.29 | 9.35％ | 183.15 | 8.26％ |
| 1/4H+3/4L | 152.41 | 152.02 | 148.08 | 152.22 | 2.79％ | 150.27 | 1.29％ |
| 1/8H+7/8L | 126.9 | 132.61 | 131.45 | 129.76 | -1.29％ | 133.83 | -3.04％ |
| 1/16H+15/16L | 125.73 | 123.87 | 123.14 | 124.80 | 1.35％ | 125.61 | -0.64％ |
| L | 114.10 | 115.57 | 114.83 | 114.83 | 0.00％ | 117.39 |
Note that: the relative deviation 1 is the relative deviation of the mean value from the theoretical value, and the relative deviation 2 is the relative deviation of the mean value from the estimated value.
As can be seen from the experimental results, the kit (group A) prepared in the embodiment 1 of the present invention has a relative deviation 1 between the detection value and the theoretical value and a relative deviation 2 between the detection value and the estimated value in the linear range of the sample concentration of 114-283mg/L, which are smaller than those of the kit (group B) and the kit (group C). Meanwhile, carrying out correlation analysis on detection results of the three groups of kits and a theoretical value of sample concentration (as shown in the accompanying figures 1-3): the correlation between the detection value of the kit of the group A and the theoretical value is obviously better than that of the kit of the group B and the group C, wherein the correlation R between the detection value of the group A and the theoretical value 2 0.9997, group B R 2 0.9893, group c 0.9783. The experimental results show that the kit (group A) prepared in the embodiment 1 of the invention has better linear correlation, i.e. better quantifiability, in the linear range than the kit (group B) and the kit (group C).
Example 4 Effect of organic solvents on the Linear Range of the kit
(1) To verify the effect of adding organic solvent to the reagent R1 on the linear range of the kit, a total of 4 sets of kits were set up:
group A: the kit prepared in the embodiment 2 of the invention;
group B: the kit differs from the kit of example 2 only in that no organic solvent is added to the reagent R1;
group C: the kit is different from the kit of example 2 only in that the organic solvent added in the reagent R1 is DMF (N, N-dimethylformamide);
group D: the kit is different from the kit of the embodiment 2 only in that the organic solvent added in the reagent R1 is methanol;
and selecting a clinical ultrahigh value sample and a low value sample, then configuring each concentration gradient sample proportionally by utilizing the high value sample and the low value sample, respectively testing the samples by using the four groups of kits, respectively repeatedly measuring each sample for 2 times, reading signals by a full-automatic biochemical analyzer (Hitachi 7180), and respectively calculating measurement mean values to perform linear range investigation. The test results are shown in the following table:
table 6 test kit linear range verification
Note that: the relative deviation 1 is the relative deviation of the mean value from the theoretical value.
As shown by the experimental results, the relative deviation between the detection value and the theoretical value of the group A, the group C and the group D in the detection linear range is smaller than that of the group B, and the detection result of the group A is optimal. Meanwhile, carrying out correlation analysis on the detection results of the 4 groups of kits and the theoretical value of the concentration of the sample (as shown in the accompanying figures 4-7): correlation R between group A test detection value and theoretical value 2 0.9997, group B R 2 0.9774, group C R 2 0.9992, group D R 2 0.9976. Experimental results show that the invention can remarkably improve the detection quantifiability of the TSGF detection kit in the linear range by adding the organic solvent (especially DMSO) into the TSGF detection kit reagent R1.
(2) To verify the effect of the amount of organic solvent added to reagent R1 on the linear range of the kit, 4 sets of experiments were set up:
group A: the kit is different from the kit of the embodiment 2 only in that the content of the DMSO (the organic solvent) in the reagent R1 is 100g/L;
group B: the kit is different from the kit of the embodiment 2 only in that the addition amount of the organic solvent DMSO in the reagent R1 is 200g/L;
group C: the kit is different from the kit of the embodiment 2 only in that the addition amount of the organic solvent DMSO in the reagent R1 is 600g/L;
group D: the kit differs from the kit of example 2 only in that the amount of the organic solvent DMSO added in the reagent R1 is 1000g/L.
And selecting a clinical ultrahigh value sample and a low value sample, then configuring each concentration gradient sample proportionally by utilizing the high value sample and the low value sample, respectively testing the samples by using the three groups of kits, respectively repeatedly measuring each sample for 2 times, reading signals by a full-automatic biochemical analyzer (Hitachi 7180), and respectively calculating measurement mean values to perform linear range investigation. The test results are shown in the following table:
table 7 test kit linear range verification
Note that: the relative deviation 1 is the relative deviation of the mean value from the theoretical value.
Carrying out correlation analysis on the detection results of the 4 groups of kits and the theoretical value of the concentration of the sample (as shown in the accompanying figures 8-11): correlation R between group A test detection value and theoretical value 2 0.9887, group B R 2 0.9971, group C R 2 0.9996, group D R 2 The experimental result shows that the invention can remarkably improve the metering property of the detection in the linear range of the kit by adding 200-600g/L of organic solvent into the reagent R1, wherein the metering property is 0.9960.
Example 5 Effect of surfactant and inorganic salt composition on kit precision
(1) Influence of surfactant and inorganic salt composition on detection precision
In order to verify the influence of the addition of the surfactant and inorganic salt composition to the detection precision of the kit in the reagent R1, 8 sets of experiments were set up in this example, wherein each set of experiments uses the kit differing from example 1 only in whether the surfactant 1Triton X-100, the surfactant 2Tween-20 and the inorganic salt NaCl were added to the reagent R1, and the preparation methods of the rest of the kits were the same as in example 1. Meanwhile, the 8 groups of kits are adopted to carry out precision detection of the median sample, and the detection results are shown in the following table:
TABLE 8
Note that: "" means inclusion of the component; "×" indicates that this component is not included.
Experimental results show that when the surfactant 1TritonX-100, the surfactant 2Tween-20 and the inorganic salt NaCl are added into the reagent R1 at the same time, the detection precision of the kit is higher.
(2) Influence of surfactant and inorganic salt concentration
In order to verify the influence of the addition of the surfactant and inorganic salt composition to the linear range of the kit in the reagent R1, 5 sets of experiments were set up in this example, wherein each set of experiments was different from example 1 only in the concentration of surfactant 1Triton X-100, surfactant 2Tween-20 and inorganic salt NaCl in the reagent R1, and the preparation methods of the rest of the kits were the same as in example 1. Meanwhile, the five kits are adopted to carry out precision detection of the median sample, and the detection results are shown in the following table:
TABLE 9
Experimental results show that the detection precision of the kit is higher when the concentration range of the surfactant 1 in the reagent R1 is 5-20g/L, the concentration range of the surfactant 2 is 0.1-1g/L and the concentration range of the inorganic salt is 1-5 g/L.
(3) Influence of surfactant and inorganic salt species
In this example, 5 experiments were set up, and the kit used in each experiment was different from that of example 1 only in the kinds of surfactant 1, surfactant 2 and inorganic salt in the reagent R1, and the preparation methods of the rest of the kits were the same as those of example 1. Meanwhile, the five kits are adopted to carry out precision detection of the median sample, and the detection results are shown in the following table:
table 10
The experimental results show that the detection precision of the kit is higher when the 7 combinations are adopted in the surfactant 1, the surfactant 2 and the inorganic salt in the reagent R1, and particularly, the detection precision is highest when the surfactant 1 is Triton X-100 or Triton X-305, the surfactant 2 is LS-114 or Tween-20 and the inorganic salt is NaCl.
In summary, according to the invention, a certain amount of organic solvent (DMSO) is added to the detection reagent R1, so that the forward progress of the ninhydrin color reaction can be effectively promoted, the color reaction is close to complete reaction, the quantifiability of the detection reagent to the object to be detected in the linear range is remarkably improved, and meanwhile, the problems of sample precipitation and detection precision reduction are brought along with the addition of the organic solvent. Furthermore, the invention solves the problems of subsequent sample precipitation and precision reduction caused by adding the surfactant and salt composition into the reagent R1, and effectively improves the detection performance of the kit.
Although embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives, and variations may be made to the above embodiments by those skilled in the art without departing from the spirit and principles of the invention.
Claims (10)
1. A kit for determining a specific growth factor, comprising reagent R1 and reagent R2, characterized in that: the reagent R1 comprises a buffer solution, a surfactant 1, a surfactant 2, inorganic salts and an organic solvent; the reagent R2 comprises a color developing agent;
the surfactant 1 is at least one of Triton X-100 and Triton X-305 with the concentration of 2-20 g/L; the surfactant 2 is at least one selected from LS-114 and Tween-20 with the concentration of 0.1-0.5 g/L; the inorganic salt is at least one of sodium chloride and potassium chloride with the concentration of 1-5g/L; the organic solvent is DMSO of 200-600 g/L; the color developing agent is ninhydrin.
2. The kit for determining specific growth factors according to claim 1, wherein the reagent R2 further comprises a buffer and a preservative.
3. A kit for determining specific growth factors according to claim 2, wherein: the content of the organic solvent is 400-600g/L.
4. A kit for determining specific growth factors according to claim 3, wherein: the content of the surfactant 1 is 5-20g/L.
5. The kit for determining a specific growth factor according to any one of claims 1 to 4, wherein the buffer in the reagent R1 is at least one selected from the group consisting of 4-hydroxyethylpiperazine ethanesulfonic acid, tris buffer and GOOD' S buffer.
6. The kit for determining a specific growth factor according to claim 5, wherein the buffer in the reagent R1 is contained in an amount of 50mM to 150mM.
7. The kit for determining a specific growth factor according to any of claims 2 to 4, wherein the buffer in reagent R2 is at least one selected from the group consisting of phosphate buffer, GOOD' S buffer and succinic acid buffer, and the preservative is at least one selected from the group consisting of sodium azide and Proclin 300.
8. The kit for determining a specific growth factor according to claim 7, wherein the buffer solution in the reagent R2 is 30-50mM and the preservative is 0.1-1g/L.
9. A kit for determining a specific growth factor according to any of claims 1 to 4, wherein the chromogenic agent is present in an amount of 10 to 100mM.
10. A composition effective to enhance the detection of biochemical agents, said composition comprising: surfactant 1, surfactant 2, inorganic salt and organic solvent;
the surfactant 1 is at least one of Triton X-100 and Triton X-305 with the concentration of 2-20 g/L; the surfactant 2 is at least one selected from LS-114 and Tween-20 with the concentration of 0.1-0.5 g/L; the inorganic salt is at least one of sodium chloride and potassium chloride with the concentration of 1-5g/L; the organic solvent is DMSO of 200-600 g/L; the biochemical reagent is a detection reagent based on an ninhydrin chemochromic method.
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| CN202110917705.XA CN113740539B (en) | 2021-08-11 | 2021-08-11 | Kit for determining specific growth factors |
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| CN113740539B true CN113740539B (en) | 2023-07-25 |
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| US4274833A (en) * | 1979-10-18 | 1981-06-23 | Pickering Michael V | Ninhydrin reagent for use in amine and amino acid analyses |
| EP0431169A4 (en) * | 1989-01-19 | 1993-03-10 | Teijin Limited | Immunoassay method, reagent and kit |
| WO2001077338A2 (en) * | 2000-04-10 | 2001-10-18 | Sugen, Inc. | Human protein kinases and protein kinase-like enzymes |
| JP2006514915A (en) * | 2002-03-01 | 2006-05-18 | ダイアックス、コープ | KDR and VEGF / KDR binding peptides and their use in diagnosis and therapy |
| TW201134828A (en) * | 2004-04-02 | 2011-10-16 | Osi Pharm Inc | 6,6-bicyclic ring substituted heterobicyclic protein kinase inhibitors |
| CN102967571A (en) * | 2012-12-24 | 2013-03-13 | 北京利德曼生化股份有限公司 | Liquid single reagent for detecting alpha-L-fucosidase and preparation method thereof |
| CN104515768B (en) * | 2014-08-29 | 2017-01-25 | 湖南新大陆生物技术有限公司 | Tumor specific growth factor detection kit |
| CN106053444A (en) * | 2016-04-28 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | A kit for measuring a specific growth factor and a preparing method thereof |
| CN109425580A (en) * | 2017-08-28 | 2019-03-05 | 江苏福隆医疗器械有限公司 | A kind of tumor specific growth factor detection kit and its application method |
| CN109580504B (en) * | 2018-10-19 | 2021-10-26 | 东软威特曼生物科技(南京)有限公司 | Lipoprotein cholesterol determination reagent and kit |
| CN109799331A (en) * | 2019-03-18 | 2019-05-24 | 湖南海源医疗科技股份有限公司 | A kind of Tumor specific factor TSGF analysis strip |
| CN111721934B (en) * | 2020-06-24 | 2023-05-09 | 湖南新大陆生物技术有限公司 | Improved specific growth factor detection kit and application thereof |
| CN112858690B (en) * | 2021-01-21 | 2023-11-10 | 宁波职业技术学院 | Urine albumin/urine creatinine composite quality control product and preparation method thereof |
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