CN113687087A - Parathyroid hormone rapid detection card and preparation method thereof - Google Patents
Parathyroid hormone rapid detection card and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a parathyroid hormone rapid detection card and a preparation method thereof, wherein the detection card comprises a bottom plate, and a blood filtering pad, a combination pad, a nitrocellulose membrane and a water absorption pad which are arranged on the bottom plate; the blood filter pad is prepared after being treated by pretreatment liquid; a first anti-PTH antibody marked by a fluorescent microsphere and a chicken IgY antibody marked by fluorescence are adsorbed on the binding pad, wherein the first anti-PTH antibody is biotin and an anti-biotin-marked anti-human PTH monoclonal antibody; the nitrocellulose membrane is provided with a detection line and a quality control line, and the detection line is coated with a PTH coated antibody; the quality control line is coated with a rabbit anti-chicken IgY antibody. The invention can directly and rapidly detect whole blood, serum and plasma samples, has simple structure, is small and portable, adopts biotin and anti-biotin labeled anti-human PTH monoclonal antibody as anti-PTH first antibody, improves the stability and high efficiency of the marker, can avoid the influence of the steric hindrance of the antibody, and improves the sensitivity, specificity and accuracy.
Description
Technical Field
The invention relates to the technical field of parathyroid hormone detection, in particular to a parathyroid hormone rapid detection card and a preparation method thereof.
Background
Parathyroid hormone (PTH), a basic single-chain polypeptide hormone secreted by parathyroid chief cells, has its primary function of regulating calcium and phosphorus metabolism in vertebrates, promoting elevation of blood calcium levels and lowering blood phosphorus levels. PTH contributes to elevated plasma calcium concentrations and the major target organs of action are bone and kidney. It mobilizes bone calcium to enter blood, promotes reabsorption of calcium ions and excretion of phosphate by renal tubules, and increases blood calcium concentration and decreases blood phosphorus concentration. PTH assays can help judge parathyroid function. The PTH can be detected by chemiluminescence immunoassay, radioimmunoassay and enzyme-linked immunoassay.
However, the whole-segment parathyroid hormone detection kit mainly takes imported products as main materials, and the products are mostly enzyme-linked immunosorbent assay (ELISA) and chemiluminescence assay (CLIA). Enzyme-linked immunosorbent assay (ELISA) can only detect serum or plasma, and cannot detect whole blood. And the operation is complicated, the sample pretreatment steps are more, and the detection time is longer. The detection instruments required by the chemiluminescence method (CLIA) are large instruments, the price is high, the universality is poor, the detection can be carried out only by the sample volume of more than 200 mu L, and the matched reagent needs to be stored at 2-8 ℃, so that the requirement of bedside rapid detection cannot be met.
Therefore, the chinese invention patent CN109188000A provides a portable test strip for detecting human parathyroid hormone and a preparation method thereof, comprising a strip-shaped nitrocellulose membrane pad horizontally arranged in a transverse direction, wherein the nitrocellulose membrane pad is longitudinally spaced and mutually parallel and is respectively provided with a strip-shaped detection line and a strip-shaped quality control line, one end of the nitrocellulose membrane pad close to the quality control line is abutted with a strip-shaped water absorption pad horizontally arranged in a transverse direction, one end of the nitrocellulose membrane close to the detection line is abutted with a strip-shaped sample pad horizontally arranged in a transverse direction, the combination pad contains an anti-human PTH antibody labeled by colloidal gold or fluorescent microspheres, the detection line is coated with a detection antibody specifically binding with PTH, and the quality control line is coated with a quality control antibody specifically binding with free anti-human PTH.
However, in the scheme, the anti-human PTH antibody marked by colloidal gold or fluorescent microspheres is combined with PTH, so that the detection card has poor detection sensitivity and stability, and in the test process, because PTH is combined with the anti-human PTH antibody, a steric hindrance effect is easily caused, and the accuracy of a detection result is influenced.
In view of this, there is an urgent need to improve the test strip for absorbing and detecting human parathyroid hormone and the preparation method thereof, so as to improve the accuracy and the detection sensitivity of the detection result of the PTH detection test strip.
Disclosure of Invention
The invention aims to solve the technical problems of low detection stability and sensitivity and poor accuracy of a measurement result of the conventional PTH detection test paper.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a parathyroid hormone rapid detection card comprises a bottom plate, and a blood filtering pad, a combination pad, a nitrocellulose membrane and a water absorption pad which are arranged on the bottom plate, wherein the blood filtering pad, the combination pad, the nitrocellulose membrane and the water absorption pad are sequentially lapped on the bottom plate;
the blood filter pad is prepared after being treated by pretreatment liquid;
a first anti-PTH antibody marked by a fluorescent microsphere and a fluorescently-marked chicken IgY antibody are adsorbed on the binding pad, wherein the first anti-PTH antibody is biotin and an anti-biotin-marked anti-human PTH monoclonal antibody;
the nitrocellulose membrane is provided with a detection line and a quality control line, the detection line is arranged at one end, close to the binding pad, of the nitrocellulose membrane, and the detection line is coated with a PTH coated antibody; the quality control line is arranged on one end, close to the water absorption pad, of the nitrocellulose membrane, and the rabbit anti-chicken IgY antibody is coated on the quality control line.
In the above scheme, preferably, the pretreatment solution is 0.1-1.2% Tris buffer solution, and contains 0.01-0.4 mg/ml mouse anti-RBC antibody, 0.1-1.5 mg/ml blocking agent, and 0.1-1% surfactant, and the pH value is 7.4.
In the above protocol, preferably, the pretreatment solution is 0.1M Tris buffer containing 0.08mg/ml of mouse anti-RBC antibody, 0.4mg/ml of blocking agent, 1% S17 and 1% Tween 20, and has a pH of 7.4.
In the above scheme, preferably, the bottom plate is a PVC plate, and the blood filter pad is made of a dense glass cellulose membrane.
In the above scheme, preferably, the bottom surface of the bottom plate is provided with a two-dimensional code and a detection card code for recording and querying a detection result of the parathyroid hormone rapid detection card.
In the above scheme, preferably, the quality control antibody is a rabbit anti-chicken IgY antibody.
A preparation method of a parathyroid hormone detection card comprises the following steps:
taking a fluorescent microsphere-labeled anti-PTH first antibody and a fluorescent microsphere-labeled chicken IgY antibody, and spraying the fluorescent microsphere-labeled anti-PTH first antibody and the fluorescent microsphere-labeled chicken IgY antibody on a glass fiber membrane to prepare a binding pad, wherein the anti-PTH first antibody is biotin and an anti-biotin-labeled anti-human PTH monoclonal antibody;
coating a PTH coated antibody on a nitrocellulose membrane as a detection line, and coating a rabbit anti-chicken IgY antibody on the nitrocellulose membrane as a quality control line;
and (3) treating the blood filtering pad by using pretreatment liquid, and sequentially adhering the blood filtering pad, the combining pad and the water absorbing pad on the bottom plate to obtain the parathyroid hormone rapid detection card.
In the above embodiment, preferably, the spraying amount of the fluorescent microsphere labeled anti-PTH primary antibody is 4. mu.L/cm, and the spraying amount of the fluorescent microsphere labeled chicken IgY antibody is 4. mu.L/cm.
In the above embodiment, preferably, the preparation of the fluorescent microsphere-labeled anti-PTH first antibody comprises the following steps:
adding the activated fluorescent microspheres into an anti-biotin antibody, and reacting at room temperature in a dark place to obtain a fluorescence-labeled anti-biotin antibody;
adding biotin into a buffer solution containing an anti-human PTH monoclonal antibody, and reacting at room temperature in a dark place to obtain the biotin-labeled anti-human PTH monoclonal antibody;
and mixing the biotin-labeled anti-human PTH monoclonal antibody with the obtained fluorescence-labeled antibiotic antibody, and reacting at room temperature in a dark place to obtain the fluorescence microsphere-labeled anti-biotin antibody and the biotin-labeled anti-human PTH monoclonal antibody.
In the scheme, preferably, the coating concentration of the PTH coating antibody on the detection line is 0.5-2 mg/ml, and the spraying amount is 1 mu L/cm; the coating concentration of the rabbit anti-chicken IgY antibody on the quality control line is 0.2-1 mg/ml, and the spraying amount is 1 mu L/cm.
Compared with the prior art, the parathyroid hormone rapid detection card and the preparation method thereof provided by the invention have the following advantages:
(1) the whole blood, serum and plasma sample can be directly and quickly detected, the detection can be realized only by simply processing the sample, the detection result can be quickly issued in a short time, and the detection efficiency is greatly improved;
(3) the structure is simple, the device is small and portable, a large detection machine is not required, and the requirements of on-site (such as bedside) quick detection or POCT (Point-of-care testing) are met;
(4) the detection card can be stored at the temperature of 2-30 ℃, low-temperature storage is not needed, the limitation on the storage condition of the detection card is reduced, and the application range of the detection card is expanded;
(5) the biotin and the anti-biotin-labeled anti-human PTH monoclonal antibody are used as the anti-PTH first antibody, in the preparation process, the fluorescent microsphere is combined with the anti-biotin, the biotin is combined with the anti-human PTH monoclonal antibody, and then the anti-biotin antibody is combined with the specificity of the biotin, so that the stability and the high efficiency of the marker are improved, the influence of the steric hindrance of the antibody can be avoided, and the sensitivity, the specificity and the accuracy are improved.
Drawings
FIG. 1 is a schematic structural diagram of a rapid parathyroid hormone detection card according to the present invention;
FIG. 2 is a schematic diagram showing the detection result of the rapid parathyroid hormone detection card in example 2 of the present invention;
FIG. 3 shows the results of the Roche electrochemiluminescence assay of the sample of example 2.
Detailed Description
The invention provides a parathyroid hormone rapid detection card and a preparation method thereof, which adopt a double-antibody sandwich principle and adopt biotin and an anti-biotin-labeled anti-human PTH monoclonal antibody as an anti-PTH first antibody, thereby greatly improving the accuracy and stability of parathyroid hormone detection. The invention is described in detail below with reference to the drawings and the detailed description.
As shown in figure 1, the rapid parathyroid hormone detection card and the preparation method thereof provided by the invention comprise a bottom plate 1, and a blood filtering pad 2, a combination pad 3, a nitrocellulose membrane 4 and an absorbent pad 5 which are sequentially lapped on the bottom plate 1.
The base plate 1 is a PVC plate, PVC is a short name for polyvinyl chloride, and is a polymer obtained by polymerizing vinyl chloride monomer in the presence of initiators such as peroxides and azo compounds or in the presence of light and heat according to a radical polymerization mechanism. PVC has good fireproof and heat-resistant functions, physical and chemical stability and ductility, and is widely applied to disposable medical appliance products. The bottom surface of the bottom plate 1 is provided with a two-dimensional code and a detection card code which are used for recording and inquiring the detection result of the parathyroid hormone rapid detection card.
The blood filtering pad 2 is made of a compact glass cellulose membrane, and the blood filtering membrane can quickly separate plasma in whole blood without hemolysis, so that a tester does not need to centrifuge the blood into serum when using the blood filtering pad, the testing time is saved, the efficiency is increased, peripheral blood or fresh blood can be directly used at the moment of testing, and the portability and the use convenience of the testing paper are improved. The blood filtering pad 2 is prepared after being treated by a pretreatment solution, the pretreatment solution is 0.1-1.2% Tris buffer solution, and the pretreatment solution comprises 0.01-0.4 mg/ml mouse anti-RBC antibody, 0.1-1.5 mg/ml blocking agent and 0.1-1% surfactant, and the pH value of the solution is 7.4.
In another embodiment of the invention, the pretreatment solution is 0.1M Tris buffer containing 0.08mg/ml murine anti-RBC antibody, 0.4mg/ml blocking agent, 1% S17 and 1% Tween 20 at pH 7.4.
The binding pad 3 is adsorbed with a fluorescent microsphere labeled anti-PTH primary antibody and a fluorescent labeled chicken IgY antibody, wherein the anti-PTH primary antibody is biotin and anti-biotin labeled anti-human PTH monoclonal antibody, and the binding pad 3 is used for binding PTH and the anti-PTH primary antibody.
The nitrocellulose membrane 4 is provided with a detection line 41 and a quality control line 42, the detection line 41 is arranged at one end, close to the binding pad 3, of the nitrocellulose membrane 4, and the detection line 41 is coated with a PTH (PTH) coating antibody; the quality control line 42 is arranged on one end of the nitrocellulose membrane 4 close to the absorbent pad 5, and the rabbit anti-chicken IgY is coated on the quality control line 42.
The absorbent pad 5 is located at the end of the test card and has the main function of being used as a test paper slot for absorbing the detection sample flowing in the chromatography to prevent the sample after reaction from flowing back in the reverse direction, thereby preventing the occurrence of false positive when the test report is read late due to the reverse flow. The absorbent pad is made of hydrophilic material with strong water absorption capacity and large water absorption volume, and has the characteristics of high absorption rate, high strength and high stability.
The parathyroid hormone rapid detection card is constructed based on a double-antibody sandwich principle, and parathyroid hormone (PTH) in a sample to be detected, a fluorescent microsphere-labeled antibiotic antibody on a binding pad and a biotin-labeled PTH monoclonal antibody form a compound; when passing through the detection line, the complex is combined with the PTH coated antibody on the detection line to form a double-antibody sandwich complex of a fluorescent microsphere-labeled anti-biotin antibody and a biotin-labeled anti-PTH first antibody-PTH-PTH coated antibody. The color development degree of the strip on the detection line is in direct proportion to the PTH concentration in the sample to be detected, and the PTH content in the sample to be detected can be quantitatively detected.
The present invention will be described in detail with reference to the following embodiments.
Example 1
Preparation of (I) bonding pad
(1) Washing a proper amount of hydroxyl-modified fluorescent microspheres with MES buffer solution with the value of 0.1M, pH being 5.0, centrifuging, discarding the supernatant, re-suspending with 1mL of MES buffer solution with the value of 0.1M, pH being 5.0, adding EDC to the final concentration of 10mM and adding NHS to the final concentration of 10mM, and activating in the dark at room temperature for 30 minutes to obtain activated fluorescent microspheres;
(2) washing the activated fluorescent microspheres with 100mM MES buffer solution with pH value of 5.0, adding 50 μ L of 1mg/ml anti-biotin antibody, reacting at room temperature in a dark place for 30 minutes, centrifuging, removing the unbound anti-biotin antibody, and resuspending to obtain the anti-biotin antibody labeled by the fluorescent microspheres;
(3) adding 50 μ L of 1mg/ml biotin dissolved in DMF into 0.05M PBS buffer solution containing 0.3mg of anti-human PTH monoclonal antibody, wherein the pH value of the PBS buffer solution is 7.2; reacting the buffer solution added with biotin for 1 hour in a dark place at room temperature, dialyzing at 4 ℃ overnight, mixing the buffer solution with the fluorescent microsphere-labeled anti-biotin antibody obtained in the step (2), reacting for 2 hours in a dark place at room temperature, and adding BSA with the final concentration of 1% for sealing; after the blocking is finished, washing and resuspending the antibody by using PBS (phosphate buffer solution) with the value of 0.05M, pH being 7.2 to obtain a first anti-PTH antibody marked by the fluorescent microspheres, and storing the antibody at the temperature of 4 ℃ for later use;
(4) and (4) spraying the fluorescent microsphere-labeled anti-PTH first antibody obtained in the step (3) onto a glass fiber membrane, wherein the spraying amount is 4 mu L/cm, and drying at 37 ℃ overnight to obtain the bonding pad.
Preparation of blood filtering pad
The blood filter pad was obtained by drying overnight in a dry box at 37 ℃ with a pretreatment solution (pretreated glass fiber membrane, wherein the pretreatment solution was 0.1M Tris buffer containing 1% surfactant and the pH of the Tris buffer was 7.4) containing 0.08mg/ml mouse anti-RBC antibody and 0.4mg/ml blocking agent.
Preparation of quick detecting card for (III) parathyroid hormone
(1) Preparation of nitrocellulose membrane (NC membrane) coated with detection line (T line) and quality control line (C line):
PTH-coated antibody (T line antibody) was diluted to 1mg/ml and rabbit anti-chicken IgY antibody (C line antibody) was diluted to 0.8mg/ml with coating protein diluent, respectively, and the sprayed amount was set to 1. mu.L/cm under the condition of humidity of 50% to 65%, and streaked on a nitrocellulose membrane. After streaking, the mixture is dried at 50 ℃ for 2 hours and put into a sealed bag under the condition that the humidity is lower than 35% for standby.
(2) Of test cardsAssembling: sequentially adhering a treated blood filtering pad, a binding pad adsorbing a fluorescence-labeled anti-PTH first antibody, a nitrocellulose membrane coated with a detection line and a quality control line and a water absorption pad on a PVC (polyvinyl chloride) plate to obtain a large test paper plate, and cutting the large test paper plate into test paper plates according to use requirements4mmWide, and the test paper is put into a plastic card to form the parathyroid hormone rapid detection card with the structure shown in figure 1.
Example 2
The accuracy detection test for the parathyroid hormone rapid detection card comprises the following steps:
(1) the parathyroid hormone rapid detection card prepared in example 1 was used for detection using a fluorescence immunoassay analyzer.
(2) Setting parameters of the fluorescence immunoassay analyzer: after the technological parameters of the detection card are set on the fluorescence immunoassay analyzer, the assembled detection card is taken, parathyroid hormone calibrators of 0, 11.3, 35.1, 117, 225 and 994pg/mL are respectively used for measurement by the detection card, the fluorescence intensity value of each calibrator is obtained, and the result is input into the parameters of the analyzer to complete the setting of the parameters of the analyzer.
(3) The main detection materials are: the clinical samples are obtained from related hospitals, and the total number of the samples is 300 according to an electrochemiluminescence method, wherein 100 parts of serum samples, 100 parts of plasma samples, 100 parts of whole blood samples, and the distribution interval of the parathyroid hormone content is 10-2000 pg/mL.
(4) The detection method comprises the following steps:
step 1: balancing the parathyroid hormone rapid detection card and the sample to room temperature, taking out the detection card, and horizontally placing;
step 2: sample adding: adding 30 mu L of whole blood sample into the sample diluent, or adding 20 mu L of serum/plasma sample into the sample diluent, shaking and uniformly mixing, and then vertically dropwise adding 100 mu L of the mixture to the sample adding position of the detection card; taking sample without sucking bubbles; after sample adding, standing the rapid detection card for 15 min;
and step 3: reading a card: opening a reagent card detection instrument, and selecting a project file corresponding to the kit batch number; selecting a sample type, "whole blood" or "serum/plasma"; the two-dimensional code on the bottom plate 1 is inserted into a reagent card detection instrument inwards, and the result is read on the instrument.
(5) And (3) analyzing test results:
after the preparation of the clinical sample detection reagent is finished, all clinical samples are detected according to a detection method, and detection results are analyzed.
(6) And (3) test results:
the detection results of some samples are shown in fig. 2, and a scatter plot is drawn with the detection value of the experimental system as the Y axis and the test value of the control system as the X axis, and correlation analysis is performed. The detection of the clinical samples comprises the steps of detecting 300 clinical constant value samples, wherein the average deviation value of the samples is less than 10%, the maximum deviation is less than 20%, and R2 is greater than 0.92.
The sample was detected using a Roche electrochemiluminescence kit, and the detection results are shown in FIG. 3. As shown in FIG. 3, the PTH sensitivity was 8.9pg/ml with a linear range of 8.9-1051 pg/ml.
Compared with the graph 2 and the graph 3, the result shows that the detection result of the parathyroid hormone rapid detection card provided by the invention has no obvious difference with the concentration of a fixed value sample, has high accuracy, is suitable for clinical detection, and meets the differentiation requirements of different clients on different detection occasions. Through detection, the rapid parathyroid hormone detection card provided by the invention has the advantages that the indexes such as result coincidence rate, detection limit, specificity and the like are all larger than 90%, the false detection rate of positive patients and normal people is low, and the accuracy of detection results is high.
The detection result of the PTH detection card in the prior art is easy to deviate due to the space position. Steric hindrance effect, also known as steric effect, refers to steric hindrance caused by some atoms or groups in molecules approaching each other, and the atoms or groups in molecules close to the reaction center occupy a certain spatial position, which may affect the reactivity of the molecules. In the detection process, because the antibody is combined with the antigen and is influenced by the steric hindrance effect, the structure for combining the antigen and the antibody is possibly buried in a large protein structure, so that the antigen and the antibody are prevented from being fully combined, and the accuracy of detection data is influenced.
Compared with the prior art, the parathyroid hormone rapid detection card provided by the invention introduces biotin and an anti-biotin antibody, utilizes the combination of fluorescent microspheres and anti-biotin, biotin and anti-human PTH monoclonal antibody, and then utilizes the specific combination of the anti-biotin antibody and biotin, relieves the steric hindrance effect existing in the detection process, improves the stability and high efficiency of a marker, and thus improves the accuracy of the parathyroid hormone detection card detection result.
The invention can directly and rapidly detect whole blood, serum and plasma samples without complex treatment of the samples, can rapidly issue detection results in a short time, has simple structure, is small and portable, greatly improves the PTH hormone detection efficiency, and can be widely applied to wards and postoperative PTH detection. The reagent card detecting instrument reads the two-dimensional code on the bottom plate, can synchronize the detection result with the detection card code, and is convenient to inquire the detection result.
The present invention is not limited to the above-mentioned preferred embodiments, and any structural changes made under the teaching of the present invention shall fall within the scope of the present invention, which is similar or similar to the technical solutions of the present invention.
Claims (10)
1. A parathyroid hormone rapid detection card is characterized by comprising a bottom plate, and a blood filtering pad, a combination pad, a nitrocellulose membrane and a water absorption pad which are arranged on the bottom plate, wherein the blood filtering pad, the combination pad, the nitrocellulose membrane and the water absorption pad are sequentially overlapped on the bottom plate;
the blood filter pad is prepared after being treated by pretreatment liquid;
a first anti-PTH antibody marked by a fluorescent microsphere and a fluorescently-marked chicken IgY antibody are adsorbed on the binding pad, wherein the first anti-PTH antibody is biotin and an anti-biotin-marked anti-human PTH monoclonal antibody;
the nitrocellulose membrane is provided with a detection line and a quality control line, the detection line is arranged at one end, close to the binding pad, of the nitrocellulose membrane, and the detection line is coated with a PTH coated antibody; the quality control line is arranged on one end, close to the water absorption pad, of the nitrocellulose membrane, and the rabbit anti-chicken IgY antibody is coated on the quality control line.
2. The parathyroid hormone rapid detection card of claim 1, wherein the pretreatment solution is 0.1-1.2% Tris buffer solution, and comprises 0.01-0.4 mg/ml mouse anti-RBC antibody, 0.1-1.5 mg/ml blocker, 0.1-1% surfactant, and has pH of 7.4.
3. The rapid parathyroid hormone detection card of claim 1, wherein the pretreatment solution is 0.1M Tris buffer solution containing 0.08mg/ml of mouse anti-RBC antibody, 0.4mg/ml of blocking agent, 1% S17 and 1% Tween 20, and has pH of 7.4.
4. The rapid parathyroid hormone test card of claim 1, wherein the base plate is a PVC plate and the hemofilter pad is made of a dense glass cellulose membrane.
5. The parathyroid hormone rapid detection card of claim 1, wherein a two-dimensional code and a detection card code are provided on the bottom surface of the bottom plate for recording and querying a detection result of the parathyroid hormone rapid detection card.
6. The rapid parathyroid hormone test card of claim 1, wherein the quality control antibody is rabbit anti-chicken IgY antibody.
7. A preparation method of a parathyroid hormone rapid detection card is characterized by comprising the following steps:
taking a fluorescent microsphere-labeled anti-PTH first antibody and a fluorescent microsphere-labeled chicken IgY antibody, and spraying the fluorescent microsphere-labeled anti-PTH first antibody and the fluorescent microsphere-labeled chicken IgY antibody on a glass fiber membrane to prepare a binding pad, wherein the anti-PTH first antibody is biotin and an anti-biotin-labeled anti-human PTH monoclonal antibody;
coating a PTH coated antibody on a nitrocellulose membrane as a detection line, and coating a rabbit anti-chicken IgY antibody on the nitrocellulose membrane as a quality control line;
and (3) treating the blood filtering pad by using pretreatment liquid, and sequentially adhering the blood filtering pad, the combining pad and the water absorbing pad on the bottom plate to obtain the parathyroid hormone rapid detection card.
8. The method for preparing a parathyroid hormone rapid detection card of claim 7, wherein the spraying amount of the fluorescent microsphere labeled anti-PTH first antibody is 4 μ L/cm, and the spraying amount of the fluorescent microsphere labeled chicken IgY antibody is 4 μ L/cm.
9. The rapid parathyroid hormone test card of claim 7, wherein the preparation of the fluorescent microsphere-labeled anti-PTH first antibody comprises the steps of:
adding the activated fluorescent microspheres into an anti-biotin antibody, and reacting at room temperature in a dark place to obtain a fluorescence-labeled anti-biotin antibody;
adding biotin into a buffer solution containing an anti-human PTH monoclonal antibody, and reacting at room temperature in a dark place to obtain the biotin-labeled anti-human PTH monoclonal antibody;
and mixing the biotin-labeled anti-human PTH monoclonal antibody with the obtained fluorescence-labeled antibiotic antibody, and reacting at room temperature in a dark place to obtain the fluorescence microsphere-labeled anti-biotin antibody and the biotin-labeled anti-human PTH monoclonal antibody.
10. The parathyroid hormone rapid detection card of claim 7, wherein, the coating concentration of PTH coating antibody on the detection line is 0.5-2 mg/ml, and the spraying amount is 1 μ L/cm;
the coating concentration of the rabbit anti-chicken IgY antibody on the quality control line is 0.2-1 mg/ml, and the spraying amount is 1 mu L/cm.
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| CN202110983307.8A CN113687087A (en) | 2021-08-25 | 2021-08-25 | Parathyroid hormone rapid detection card and preparation method thereof |
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| CN202110983307.8A CN113687087A (en) | 2021-08-25 | 2021-08-25 | Parathyroid hormone rapid detection card and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114184796A (en) * | 2021-12-03 | 2022-03-15 | 广州达泰生物工程技术有限公司 | Kit and method for quantitatively detecting full-segment parathyroid hormone |
| CN115184620A (en) * | 2022-09-14 | 2022-10-14 | 山东子峰生物技术有限公司 | Quantum dot fluorescence detection test strip and kit for PLA2R antibody and application of quantum dot fluorescence detection test strip and kit |
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2021
- 2021-08-25 CN CN202110983307.8A patent/CN113687087A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114184796A (en) * | 2021-12-03 | 2022-03-15 | 广州达泰生物工程技术有限公司 | Kit and method for quantitatively detecting full-segment parathyroid hormone |
| CN115184620A (en) * | 2022-09-14 | 2022-10-14 | 山东子峰生物技术有限公司 | Quantum dot fluorescence detection test strip and kit for PLA2R antibody and application of quantum dot fluorescence detection test strip and kit |
| CN115184620B (en) * | 2022-09-14 | 2023-01-24 | 山东子峰生物技术有限公司 | Quantum dot fluorescence detection test strip and kit for PLA2R antibody and application of quantum dot fluorescence detection test strip and kit |
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