CN111521832A - Kit for determining 25-hydroxy-vitamin D - Google Patents
Kit for determining 25-hydroxy-vitamin D Download PDFInfo
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- CN111521832A CN111521832A CN202010344001.3A CN202010344001A CN111521832A CN 111521832 A CN111521832 A CN 111521832A CN 202010344001 A CN202010344001 A CN 202010344001A CN 111521832 A CN111521832 A CN 111521832A
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- G—PHYSICS
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- 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
- G01N33/82—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving vitamins or their receptors
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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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
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Abstract
The invention discloses a kit for measuring 25-hydroxy-vitamin D, which comprises a first reagent R1, a second reagent R2 and a third reagent R3; the first reagent R1 comprises a magnetic bead coated by goat monoclonal antibody, biotin is marked on the magnetic bead, the second reagent R2 comprises a 25-OH VD derivative containing an alkaline phosphatase marker, and the third reagent R3 comprises a dissociation agent which is sodium acetate. The sodium acetate in the third reagent R3 can effectively separate 25-OH VD from the binding protein in the sample, thereby obviously improving the specificity and sensitivity of 25-hydroxy-vitamin D detection and further improving the accuracy of the detection result.
Description
Technical Field
The invention relates to the technical field of detection kits, in particular to a kit for determining 25-hydroxy-vitamin D.
Background
Vitamin D (vitamin D) is a cyclopentane polyhydrophenanthrene compound, which is necessary for normal bone growth and calcium and phosphorus metabolism, the main metabolite of the compound is 25-hydroxy-vitamin D, which has two forms in human body, vitamin D2 and vitamin D3, vitamin D2 is obtained from plant-derived food at least partially, vitamin D3 is the main form existing in human body, and is generated by dehydrocholesterol in skin through sunlight irradiation. The level of 25-hydroxyvitamin D in serum may reflect the storage level of vitamin D and is associated with clinical symptoms of vitamin D deficiency.
The clinical application of the detection of 25-hydroxy-vitamin D is related to the diagnosis, treatment and monitoring of rickets, osteomalacia, postmenopausal osteoporosis and renal bone diseases in children, and can be used for auxiliary diagnosis of the diseases. The existing 25-OH VD (25-hydroxy-vitamin D) immunoassay reagent often has the phenomenon of poor dissociation effect when being used for measuring clinical samples, so that the detection result is inaccurate, and the clinical use is influenced.
Disclosure of Invention
The invention aims to provide a kit for measuring 25-hydroxy-vitamin D, which solves the problem of inaccurate detection result caused by poor dissociation effect of the existing 25-OHVD immunoassay reagent.
The invention is realized by the following technical scheme:
a kit for the determination of 25-hydroxy-vitamin D comprising a first reagent R1, a second reagent R2 and a third reagent R3;
the first reagent R1 comprises a magnetic bead coated by goat monoclonal antibody, biotin is marked on the magnetic bead, the second reagent R2 comprises a 25-OH VD derivative containing an alkaline phosphatase marker, and the third reagent R3 comprises a dissociation agent which is sodium acetate.
The sodium acetate in the third reagent R3 can effectively separate 25-OH VD from the binding protein in the sample, thereby obviously improving the specificity and sensitivity of 25-hydroxy-vitamin D detection and further improving the accuracy of the detection result.
According to the invention, the first reagent R1, the second reagent R2 and the third reagent R3 are matched with each other, so that the specificity and the sensitivity of 25-hydroxy-vitamin D detection can be improved, and furthermore, the third reagent R3 is an acidic buffer solution, and the pH value of the acidic buffer solution is 3-5.
The applicant found through experiments that: in a low pH environment, the dissociation effect of sodium acetate is better and the stability of the third reagent R3 is good, but when the pH is too low, the reaction rate of the whole system is reduced and the dissociation effect is poor.
Further, the pH value of the acidic buffer is 3.95-4.05.
The pH value is about 4, which is the optimal pH value of the third reagent R3, so that the dissociation effect is optimal and the stability is particularly good.
Further, the acidic buffer solution is adjusted to pH with glacial acetic acid.
Further, the preparation method of the acidic buffer solution comprises the following steps:
A. weighing sodium acetate and a clean container, and adding purified water for dissolving;
B. adding glacial acetic acid into the solution obtained in the step A to adjust the pH value;
C. and D, using purified water to fix the volume of the solution prepared in the step B.
Further, the goat monoclonal antibody coated magnetic beads are coated by Biotin-25-hydroxy vitamin D monoclonal antibody magnetic beads, wherein the coating amount of the magnetic beads is 1 mu g/mg, and the re-dissolution ratio is 1/50.
The coating amount means: 1 mu g of Biotin-25-hydroxy vitamin D monoclonal antibody corresponds to 1mg of magnetic beads, the coating amount has the significance of linking the antibody to the magnetic beads according to a certain proportion, and the proportion effect ensures that the reaction system is better; the dilution ratio (redissolution ratio) means: the developed naked magnetic beads are original-time, 1/50 is diluted by 50 times on the basis of original time, the dilution ratio is significant in that the original-time naked magnetic beads are too high in concentration in a reaction system and have surplus when being linked with an antibody, the excessive dilution ratio can cause deletion when the antibody is linked, and through repeated tests, the naked magnetic beads are diluted to 1/50, so that the reaction system is more sufficient, and the effect is better.
Further, the first reagent R1 consists of 1.21g/L Tris, 9.00g/L NaCl, about 0.7mL/L concentrated HCl, 10.00g/L BSA, 1mL/L PC-950, 0.5mL/L Tween-20, 0.2. mu.g/mL goat anti-human 25-OH VD specific monoclonal antibody, 0.2mg/mL streptavidin magnetic bead and 50ul/mg magnetic bead blocking solution.
The preparation method of the magnetic bead sealing solution comprises the following steps: D-Biotin is placed at room temperature (protected from light) and is balanced to room temperature (10-30 min); taking a piece of clean weighing paper, placing the weighing paper on a balance, and adjusting a zero point; D-Biotin 244.5mg was weighed into an appropriate container and dissolved thoroughly in 50mL DMSO, 50mL glycerol was added, mixed thoroughly, stored at-20 ℃ and 50uL blocking solution added to 1mg magnetic beads.
Further, a second reagentR2 is composed of 1.21g/L Tris, 9.00g/L NaCl, about 0.7mL/L concentrated HCl, 10.00g/L BSA, 1mL/L PC-950, 0.5mL/L Tween-20, 10 mL/L0.1M MgCl2、10mL/L 0.01M ZnCl2And 1 mg/. mu.g of an alkaline phosphatase-activated 25-OH VD derivative.
Further, the pH values of the first reagent R1 and the second reagent R2 are both 7.35-7.45.
Further, the reaction system of the first reagent R1, the second reagent R2 and the first reagent R3 was added in an amount of 1:1: 1.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the kit comprises a first reagent R1, a second reagent R2 and a third reagent R3, wherein the first reagent R1 and the second reagent R2 are respectively sheep monoclonal antibody coated magnetic beads and enzyme working solution, the third reagent R3 contains a sodium acetate dissociation agent, the sheep monoclonal antibody coated magnetic beads and the enzyme working solution provide necessary conditions for detecting 25-hydroxy-vitamin D, and the sodium acetate dissociation agent can effectively separate 25-OH VD from binding protein in a sample, so that the specificity and the sensitivity of 25-hydroxy-vitamin D detection are remarkably improved, and the accuracy of a detection result is further improved.
2. The third reagent R3 of the kit is an acidic buffer solution, the pH value of the acidic buffer solution is 3-5, sodium acetate is used as a dissociation agent, glacial acetic acid is used for adjusting the pH value, the dissociation effect of the sodium acetate is better under the low pH value environment, and the stability of the third reagent R3 is good.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not used as limitations of the present invention.
Example 1:
a kit for the determination of 25-hydroxy-vitamin D comprising a first reagent R1, a second reagent R2 and a third reagent R3;
the first reagent R1 comprises a magnetic bead coated by goat monoclonal antibody, biotin is marked on the magnetic bead, the second reagent R2 comprises a 25-OH VD derivative containing an alkaline phosphatase marker, the third reagent R3 is an acidic buffer solution, the pH value of the acidic buffer solution is 3, the acidic buffer solution comprises a dissociation agent, and the dissociation agent is sodium acetate; the preparation method of the acidic buffer solution comprises the following steps:
A. preparing a clean container, weighing 82.03g of 1M NaAc by using a laboratory balance, and adding 4.5L of purified water for dissolving;
B. adding a proper amount of glacial acetic acid into the solution obtained in the step A, and adjusting the pH value to 2.95-3.05 by using 300mL of glacial acetic acid;
C. using purified water to fix the volume of the solution in the B to 5L to prepare 0.2mol/L sodium acetate buffer solution;
the first reagent R1 consists of 1.21g/L Tris, 9.00g/L NaCl, about 0.7mL/L concentrated HCl, 10.00g/L LBSA, 1mL/L PC-950, 0.5mL/L Tween-20, 0.2 mu g/mL sheep anti-human 25-OH VD specific monoclonal antibody, 0.2mg/mL streptavidin magnetic bead and 50ul/mg magnetic bead blocking solution; the preparation method of the magnetic bead sealing solution comprises the following steps: D-Biotin is placed at room temperature (in a dark place) and is balanced to the room temperature for 30 min; taking a piece of clean weighing paper, placing the weighing paper on a balance, and adjusting a zero point; weighing 244.5mg of D-Biotin in a proper container, fully dissolving the D-Biotin in 50mL of DMSO (dimethyl sulfoxide), adding 50mL of glycerol, fully and uniformly mixing, storing at-20 ℃, and adding 50uL of confining liquid into 1mg of magnetic beads;
the second reagent R2 consists of 1.21g/L Tris, 9.00g/L NaCl, about 0.7mL/L concentrated HCl, 10.00g/LBSA, 1mL/L PC-950, 0.5mL/L Tween-20, 10 mL/L0.1M MgCl2、10mL/L 0.01M ZnCl2And 1 mg/mug of an alkaline phosphatase-activated 25-OH VD derivative;
the pH values of the first reagent R1 and the second reagent R2 are both 7.35-7.45; the adding amount ratio of the first reagent R1 to the second reagent R2 to the first reagent R3 is 1:1: 1.
Example 2:
this example is based on example 1, and differs from example 1 in that:
the pH value of the acidic buffer solution is 4, and the preparation method of the acidic buffer solution comprises the following steps:
A. preparing a clean container, weighing 1M NaAc by using a laboratory balance, and adding purified water to dissolve;
B. adding a proper amount of glacial acetic acid into the solution obtained in the step A, and adjusting the pH value to 3.95-4.05 by using 200mL of glacial acetic acid;
C. and (5) adding purified water to the solution in the solution B to a constant volume of 5L to prepare 0.2mol/L sodium acetate buffer solution.
Example 3:
this example is based on example 1, and differs from example 1 in that:
the pH value of the acidic buffer solution is 5, and the preparation method of the acidic buffer solution comprises the following steps:
A. preparing a clean container, weighing 1M NaAc by using a laboratory balance, and adding purified water to dissolve;
B. adding a proper amount of glacial acetic acid into the solution obtained in the step A, and adjusting the pH value to 4.95-5.05 by using 100mL of glacial acetic acid;
C. and (5) adding purified water to the solution in the solution B to a constant volume of 5L to prepare 0.2mol/L sodium acetate buffer solution.
Comparative example 1:
this comparative example is based on example 1 and differs from example 1 in that:
the dissociation agent is sodium citrate.
Comparative example 2:
this comparative example is based on example 2, differing from example 2 in that:
the dissociation agent is sodium citrate.
Comparative example 3:
this comparative example is based on example 3, differing from example 3 in that:
the dissociation agent is sodium citrate.
Comparative example 4:
this comparative example is based on example 1 and differs from example 1 in that:
the third reagent R3 was not included.
The kits described in example 2, comparative example 2 and comparative example 4 were used in comparative experiments of kit sensitivity and specificity under equal conditions:
experimental materials: BNHS, 0.2mol/L PBS (PH7.4) Buffer, 1mmol/L LTris Buffer, goat anti-human 25-OHVD specific monoclonal antibody, Buffer containing 1% BSA, ALP (alkaline phosphatase) -labeled 25-OH VD derivative, dissociating agent (sodium acetate or sodium citrate or no dissociating agent), naked magnetic beads, 25-OH-VD calibrator Cal-1, Cal-6 (the concentration of Cal-1 to Cal-6 is 0, 10, 20, 40, 80, 160ng/mL), samples S1-S40, substrate solution, cleaning solution and Beckmann Access2 full-automatic chemiluminescence determinator.
The above experimental materials were prepared into the kits described in example 2, comparative example 2 and comparative example 4, respectively, and the 25-OH-VD calibrators cal-1 to cal-6 were measured, the measurement curves and specificities of the different antibodies were observed, and the samples S1-S40 were measured and compared with Rogowski values.
The reaction mode is as follows: adding 30 mu L of calibrator, 50 mu L of magnetic beads and 45 mu L of dissociating agent, incubating for 10min at 37 ℃, adding 50 mu L of enzyme working solution, incubating for 10min at 37 ℃, adding cleaning solution, cleaning and separating for three times in a magnetic field, adding substrate solution, incubating for 5min at 37 ℃, and detecting on a Beckmann Access2 full-automatic chemiluminescence determinator.
The results are shown in tables 1 and 2:
TABLE 1
TABLE 2
From the data in tables 1 and 2, it can be seen that:
the kit without the third reagent has poor sensitivity and specificity, the kit with sodium citrate as the dissociating agent has relatively poor sensitivity and specificity, the sodium citrate with different pH values can be further verified, the kit sensitivity and specificity are compared, the kit curve with sodium acetate as the dissociating agent basically meets the requirement for use, the measured sample value has little difference with the Rogowski value, and the high concentration value can be measured, so that the sensitivity is enough; sodium acetate with different pH values and a kit sensitivity and specificity comparison experiment can be further verified.
Sodium citrate with different pH values, and a kit sensitivity and specificity comparison experiment:
experimental materials: BNHS, 0.2mol/L PBS (PH7.4) Buffer, 1mmol/L tris Buffer, goat anti-human 25-OHVD specific monoclonal antibody, Buffer containing 1% BSA, ALP labeled 25-OH VD derivative, dissociation agent (buffers are sodium citrate with different PH values, PH 3, PH 4, PH 5, respectively), naked magnetic beads, 25-OH-VD calibrators Cal-1, Cal-6 (concentrations of Cal-1 to Cal-6 are 0, 10, 20, 40, 80, 160ng/mL), substrate solution, wash solution, beckmann Access2 full-automatic chemiluminescence determinator.
The experimental materials are respectively prepared into the kits of comparative examples 1 to 3, 25-OH-VD calibrators cal-1 to cal-6 are measured, and the measuring curves and specificities of different antibodies are observed.
The reaction mode is as follows: adding 30 mu L of calibrator, 50 mu L of magnetic beads and 45 mu L of dissociating agent, incubating for 10min at 37 ℃, adding 50 mu L of enzyme working solution, incubating for 10min at 37 ℃, adding cleaning solution, cleaning and separating for three times in a magnetic field, adding substrate solution, incubating for 5min at 37 ℃, and detecting on a Beckmann Access2 full-automatic chemiluminescence determinator.
The results are shown in table 3:
TABLE 3
From the data in table 3, it can be seen that:
sodium citrate is used as a dissociating agent, but due to different pH values, the kit prepared by the same labeling method has an unobvious effect, and the higher the pH value is, the lower the titer of the kit is.
Sodium acetate with different pH values, and the test for comparing the sensitivity and the specificity of the kit in examples 1 to 3, wherein the test process is the same as the test for comparing the sensitivity and the specificity of the kit with the sodium citrate with different pH values, the results are shown in Table 4:
TABLE 4
From the data in table 4, it can be seen that:
sodium acetate is used as a dissociation agent, the determination effect is better when the pH is 4, and the higher the pH is, the lower the effect of the kit is, the lower the pH is, the lower the effect of the kit is, but the effect is higher than that of an acidic buffer solution using sodium citrate as a dissociation agent.
Sodium acetate with the same pH value, and a kit sensitivity and specificity comparison experiment:
experimental materials: BNHS, 0.2mol/L PBS (PH7.4) Buffer solution, 1mmol/L LTris Buffer solution, goat anti-human 25-OHVD specific monoclonal antibody, Buffer solution containing 1% BSA, ALP-labeled 25-OH VD derivative, sodium acetate dissociator, naked magnetic beads, 25-OH-VD calibrator Cal-1, Cal-6 (the concentration of Cal-1 to Cal-6 is 0, 10, 20, 40, 80, 160ng/mL), substrate solution, cleaning solution and Beckmann Access2 full-automatic chemiluminescence determinator.
The above materials were prepared into a kit (2 boxes) as described in example 2, one portion was stored at 2-8 ℃, the other portion was stored at 37 ℃ in a constant temperature incubator, and taken out after 6 days. The 25-OH-VD calibrator was assayed by mixing the magnetic beads with a dissociation reagent (third reagent R3) and an enzyme working solution (second reagent R2), respectively. On the sixth day, the luminescence signal of the calibrator was observed with a beckmann Access2 full-automatic chemiluminescence analyzer, the signal retention rate was calculated, and the stability of sodium acetate buffered (pH 4) was verified. The results are shown in Table 5:
TABLE 5
From the data in table 5, it can be seen that:
and sodium acetate analysis buffer (pH 4) is used as a dissociation agent, so that the signal retention rate is good and the stability is better after the sample is stored at 2-8 ℃ and 37 ℃.
In conclusion, when the 25-OH-VD determination kit adopts sodium acetate as a dissociating agent, the sensitivity and specificity are effectively improved; before the sodium acetate buffer solution is reacted with biotin-coated magnetic beads as a dissociation agent, the pH value of the sodium acetate buffer solution is adjusted to 3.95-4.05 by glacial acetic acid and is adjusted to a fixed pH value, so that the stability of the kit can be improved, the effect is influenced when the pH value is lowered, resources can be wasted, and the dissociation effect is poor due to the upward pH value.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are merely exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A kit for the determination of 25-hydroxy-vitamin D, characterized by comprising a first reagent R1, a second reagent R2 and a third reagent R3;
the first reagent R1 comprises a magnetic bead coated by goat monoclonal antibody, biotin is marked on the magnetic bead, the second reagent R2 comprises a 25-OH VD derivative containing an alkaline phosphatase marker, and the third reagent R3 comprises a dissociation agent which is sodium acetate.
2. The kit for detecting 25-hydroxy-vitamin D according to claim 1, wherein the third reagent R3 is an acidic buffer solution, and the pH value of the acidic buffer solution is 3-5.
3. The kit for the assay of 25-hydroxy-vitamin D according to claim 2, wherein the pH of the acidic buffer is 3.95-4.05.
4. The kit for measuring 25-hydroxy-vitamin D according to claim 2, wherein the acidic buffer solution is adjusted in pH with glacial acetic acid.
5. The kit for measuring 25-hydroxy-vitamin D according to claim 4, wherein the preparation method of the acidic buffer comprises the following steps:
A. weighing sodium acetate and a clean container, and adding purified water for dissolving;
B. adding glacial acetic acid into the solution obtained in the step A to adjust the pH value;
C. and D, using purified water to fix the volume of the solution prepared in the step B.
6. The kit for measuring 25-hydroxy-vitamin D according to claim 1, wherein the goat monoclonal antibody coated magnetic beads are coated with Biotin-25-hydroxy vitamin D monoclonal antibody magnetic beads, the coating amount of the magnetic beads is 1 μ g/mg, and the re-dissolution ratio is 1/50.
7. The kit for detecting 25-hydroxy-vitamin D according to claim 1, wherein the first reagent R1 is composed of 1.21g/L Tris, 9.00g/L NaCl, about 0.7mL/L concentrated HCl, 10.00g/L BSA, 1mL/L PC-950, 0.5mL/L Tween-20, 0.2 μ g/mL goat anti-human 25-OH VD specific monoclonal antibody, 0.2mg/mL streptavidin magnetic beads and 50ul/mg magnetic bead blocking solution.
8. The kit for detecting 25-hydroxy-vitamin D according to claim 1, wherein the second reagent R2 is prepared from 1.21g/L Tris, 9.00g/L NaCl, about 0.7mL/L concentrated HCl, 10.00g/L BSA, 1mL/L PC-950, 0.5mL/L Tween-20, 10 mL/L0.1M MgCl2、10mL/L 0.01M ZnCl2And 1 mg/. mu.g of an alkaline phosphatase-activated 25-OH VD derivative.
9. The kit for detecting 25-hydroxy-vitamin D according to any one of claims 1 to 8, wherein the pH value of the first reagent R1 and the pH value of the second reagent R2 are both 7.35 to 7.45.
10. The kit for measuring 25-hydroxy-vitamin D according to any one of claims 1 to 8, wherein the reaction system of the first reagent R1, the second reagent R2 and the first reagent R3 is added in an amount of 1:1: 1.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117825698A (en) * | 2024-03-05 | 2024-04-05 | 江苏奥雅生物科技有限公司 | Dissociating agent for dissociating vitamin D, detection method and application |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040132104A1 (en) * | 2003-01-07 | 2004-07-08 | Sackrison James L. | Vitamin D assay |
| US20100068725A1 (en) * | 2007-02-01 | 2010-03-18 | Franz Paul Armbruster | Direct determination of vitamin d in serum or plasma |
| US20170292964A1 (en) * | 2016-04-07 | 2017-10-12 | Quidel Corporation | Processing reagent and use thereof in assays for detection of analytes associated with a binding protein |
| US20180031581A1 (en) * | 2016-07-29 | 2018-02-01 | Diazyme Laboratories, Inc. | Methods and kits for assaying a vitamin d moiety |
| CN108362688A (en) * | 2018-01-02 | 2018-08-03 | 北京利德曼生化股份有限公司 | A kind of 25(OH)VD magnetic microparticle chemiluminescence detection kit |
| CN109188003A (en) * | 2018-09-13 | 2019-01-11 | 迈克生物股份有限公司 | The method for measuring 25-hydroxy-vitamin D |
| CN109188002A (en) * | 2018-09-13 | 2019-01-11 | 迈克生物股份有限公司 | For measuring the kit of 25-hydroxy-vitamin D |
-
2020
- 2020-04-27 CN CN202010344001.3A patent/CN111521832A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040132104A1 (en) * | 2003-01-07 | 2004-07-08 | Sackrison James L. | Vitamin D assay |
| US20100068725A1 (en) * | 2007-02-01 | 2010-03-18 | Franz Paul Armbruster | Direct determination of vitamin d in serum or plasma |
| US20170292964A1 (en) * | 2016-04-07 | 2017-10-12 | Quidel Corporation | Processing reagent and use thereof in assays for detection of analytes associated with a binding protein |
| US20180031581A1 (en) * | 2016-07-29 | 2018-02-01 | Diazyme Laboratories, Inc. | Methods and kits for assaying a vitamin d moiety |
| CN109863406A (en) * | 2016-07-29 | 2019-06-07 | Diazyme研究室有限公司 | For measuring the method and ingredient of vitamin D |
| CN108362688A (en) * | 2018-01-02 | 2018-08-03 | 北京利德曼生化股份有限公司 | A kind of 25(OH)VD magnetic microparticle chemiluminescence detection kit |
| CN109188003A (en) * | 2018-09-13 | 2019-01-11 | 迈克生物股份有限公司 | The method for measuring 25-hydroxy-vitamin D |
| CN109188002A (en) * | 2018-09-13 | 2019-01-11 | 迈克生物股份有限公司 | For measuring the kit of 25-hydroxy-vitamin D |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117825698A (en) * | 2024-03-05 | 2024-04-05 | 江苏奥雅生物科技有限公司 | Dissociating agent for dissociating vitamin D, detection method and application |
| CN117825698B (en) * | 2024-03-05 | 2024-06-07 | 江苏奥雅生物科技有限公司 | Dissociating agent for dissociating vitamin D, detection method and application |
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