CN111537752A - Anti muller tube hormone detect reagent box - Google Patents
Anti muller tube hormone detect reagent box Download PDFInfo
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- CN111537752A CN111537752A CN202010420025.2A CN202010420025A CN111537752A CN 111537752 A CN111537752 A CN 111537752A CN 202010420025 A CN202010420025 A CN 202010420025A CN 111537752 A CN111537752 A CN 111537752A
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- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
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Abstract
The invention discloses an anti-mullerian hormone detection kit, which comprises: a probe substance containing a fluorescently labeled AMH antibody complex, a biotin labeled AMH antibody complex, and a fluorescently labeled quality control complex; a reaction buffer stored separately from the detection substance, the reaction buffer being capable of dissolving the detection substance. The invention also discloses a method for detecting anti-mullerian hormone by using the kit, which comprises the following steps: dissolving the detection substance in the reaction buffer solution, adding the plasma or serum to be detected, mixing and adding the mixture to the test strip. The invention prepares the AMH antibody complex, the biotin-labeled AMH antibody complex and the quality control complex coated on the binding pad into the detection substance, solves the problems of poor AMH test precision and sensitivity, and simultaneously improves the stability of the AMH kit.
Description
Technical Field
The invention belongs to the technical field of fluorescence immunochromatography in medical immunology, and relates to an anti-mullerian hormone detection kit.
Background
Currently, a common method for detecting anti-mullerian hormone (AMH) on the market is a fluorescence immunoassay method, which is based on a reagent strip with a conjugate pad, and thus, the fluorescence labeled AMH antibody complex and quality control complex bound to an antigen are directly coated on the conjugate pad of the reagent strip, as shown in fig. 1, the reagent strip includes a sample pad 1, an absorbent pad 2, a conjugate pad 3, and an NC membrane 4 (nitrocellulose membrane), the NC membrane 4 has a quality control line 5, a test line 6, and a test line 7, when a sample flows through the conjugate pad, the antigen in the sample does not sufficiently react with the coated fluorescence labeled AMH antibody complex and quality control complex because the sample flows rapidly on the NC membrane after being added to the sample pad, and when the sample flows through the conjugate pad, the sample does not sufficiently react with the antibody on the conjugate pad because the antibody is coated on the conjugate pad and is relatively incompletely exposed, resulting in deterioration of precision CV. In order to allow the sample to sufficiently react with the fluorescently labeled AMH antibody complex and the quality control complex and to secure the sensitivity of the reaction, the applicant proposed the present invention.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and/or disadvantages and to provide at least the advantages described hereinafter.
It is still another object of the present invention to provide an anti-mullerian hormone detection kit.
The technical scheme provided by the invention is as follows:
an anti-mullerian hormone detection kit, comprising:
a probe substance containing a fluorescently labeled AMH antibody complex, a biotin labeled AMH antibody complex, and a fluorescently labeled quality control complex;
a reaction buffer stored separately from the detection substance, the reaction buffer being capable of dissolving the detection substance.
Preferably, in the anti-mullerian hormone detection kit, the detection substance is a spherical particle, the detection substance includes a first detection substance and a second detection substance, and when in use, 1 particle of each of the first detection substance and the second detection substance is dissolved in 100 to 200 μ L of the reaction buffer.
Preferably, in the anti-mullerian hormone detection kit, the preparation process of the detection substance comprises:
preparing a detection substance solution: the basic buffer solution comprises 0.05-10% of BSA (bovine serum albumin) by mass percentage and 0.05-5% of sucrose by mass percentage, a first detection substance solution containing 0.01-10 mu g/mL biotin-labeled AMH antibody complex and 0.01-10 mu g/mL Eu-labeled Anti-chicken IgY antibody complex is prepared from the basic buffer solution, and then the first detection substance solution is prepared into solid particles to obtain a first detection substance; preparing a second detection substance solution containing 2-25 μ g/mL Eu-labeled AMH antibody complex by using another basic buffer solution component, and preparing the second detection substance solution into solid particles to obtain a second detection substance.
Preferably, in the anti-mullerian hormone test kit, each 1 particle of the first detection substance and the second detection substance comprises 20 to 80 μ L of the effective component of the first detection substance solution and the second detection substance solution, respectively.
Preferably, in the anti-mullerian hormone detection kit, the mass ratio of the first detection substance to the second detection substance is 0.01 to 1: 0.1-3.
Preferably, in the anti-mullerian hormone detection kit, the reaction buffer is prepared by dissolving the following components in 10mM phosphoric acidIn salt buffer: 100-600mM NaCl, 0.01-0.5% (w/v) NaN3Tween 20 of 0.1-3% (v/v), IGEPAL CA-630 of 0.01-0.5% (v/v), BSA of 0.5-3% (w/v).
Preferably, in the anti-mullerian hormone detection kit, the kit further comprises a test strip without a binding pad;
the test strip comprises a sample pad, a nitrocellulose membrane and an absorption pad which are sequentially arranged from one end of the pad card to the other end, wherein the nitrocellulose membrane is provided with a quality control line and two detection lines which are parallel to each other at intervals, and the sample pad and the absorption pad respectively press two ends of the nitrocellulose membrane.
Preferably, in the anti-mullerian tube hormone detection kit, a protective layer is coated outside the test strip, the waterproof layer comprises a waterproof silica gel cushion layer, an ethylene adsorption layer and a waterproof breathable paper layer which are sequentially arranged from inside to outside in a stacking manner, and the pore diameter of the breathable hole of the waterproof breathable paper layer is 10-15 μm.
Preferably, in the anti-mullerian hormone detection kit, the detection substance is stored under refrigeration.
The method for detecting the anti-mullerian hormone by using the kit comprises the following steps: dissolving the detection substance in the reaction buffer solution, adding the plasma or serum to be detected, mixing and adding the mixture to the test strip.
The invention at least comprises the following beneficial effects:
the invention prepares the fluorescence-labeled AMH antibody complex, the biotin-labeled AMH antibody complex and the quality control complex into the detection substance, only needs to add corresponding solvent during the reaction, can dissolve the fluorescence-labeled AMH antibody complex, the biotin-labeled AMH antibody complex and the quality control complex, at the moment, adds the sample and mixes, can make the antigen in the sample fully react with the fluorescence-labeled AMH antibody complex, the biotin-labeled AMH antibody complex and the quality control complex, and simultaneously improves the sensitivity of the reaction. The design improves the precision and the sensitivity of anti-mullerian hormone detection on one hand, and improves the stability of products because the fluorescence labeled AMH antibody complex and the quality control complex are stored in a refrigeration mode on the other hand. In addition, the outer layer of the test strip is wrapped by the protective layer, the waterproof breathable paper layer is arranged outside the protective layer, so that ethylene can be absorbed by the ethylene absorption layer (containing potassium permanganate) for example, the ethylene can be prevented from entering the interior to cause deterioration of the test strip, and meanwhile, the waterproof silica gel cushion layer is arranged inside the test strip, so that the test strip has a further waterproof effect, has a protection and anti-bumping effect, can isolate air, prolongs the service life of the test strip and prolongs the quality guarantee period of products.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
Figure 1 is a schematic representation of a prior art reagent strip for anti-mullerian hormone detection;
figure 2 is a schematic view of one embodiment of an anti-mullerian hormone test reagent strip according to the present invention;
figure 3 is a schematic representation of a reagent strip for anti-mullerian hormone detection in accordance with one embodiment of the present invention;
FIG. 4 is a diagram of the correlation between the serum detection value of the control system and the serum detection value of the experimental system in the consistency test of the kit;
FIG. 5 is a diagram of the correlation analysis between the serum test value of the test system and the plasma test value of the test system in the consistency test of the kit.
Detailed Description
The present invention is further described in detail below with reference to the attached drawings so that those skilled in the art can implement the invention by referring to the description text.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
As shown in fig. 2 and 3, the present invention provides an anti-mullerian hormone detection kit, comprising:
a probe substance containing a fluorescently labeled AMH antibody complex, a biotin labeled AMH antibody complex, and a fluorescently labeled quality control complex;
a reaction buffer stored separately from the detection substance, the reaction buffer being capable of dissolving the detection substance.
In the above aspect, preferably, the detection substance is a spherical particle, the detection substance includes a first detection substance and a second detection substance, and when in use, 1 particle of each of the first detection substance and the second detection substance is dissolved in 100 to 200 μ L of the reaction buffer.
In the above aspect, preferably, the preparation process of the detection substance includes:
preparing a detection substance solution: the basic buffer solution comprises 0.05-10% of BSA (bovine serum albumin) by mass percentage and 0.05-5% of sucrose by mass percentage, a first detection substance solution containing 0.01-10 mu g/mL biotin-labeled AMH antibody complex and 0.01-10 mu g/mL Eu-labeled Anti-chicken IgY antibody complex is prepared from the basic buffer solution, and then the first detection substance solution is prepared into solid particles to obtain a first detection substance; preparing a second detection substance solution containing 2-25 μ g/mL Eu-labeled AMH antibody complex by using another basic buffer solution component, and preparing the second detection substance solution into solid particles to obtain a second detection substance.
In the above aspect, preferably, each 1 particle of the first detection substance and the second detection substance contains 20 to 80 μ L of the effective component of the first detection substance solution and the effective component of the second detection substance solution, respectively.
In one embodiment of the present invention, it is preferable that the mass ratio of the first detection substance to the second detection substance is 0.01 to 1: 0.1-3.
In one embodiment of the present invention, preferably, the reaction buffer is prepared by dissolving the following components in 10mM phosphate buffer: 100-600mM NaCl, 0.01-0.5% (w/v) NaN3Tween 20 of 0.1-3% (v/v), IGEPAL CA-630 of 0.01-0.5% (v/v), BSA of 0.5-3% (w/v).
In one embodiment of the present invention, preferably, the kit further comprises a test strip without a binding pad;
the test strip comprises a sample pad 1, a nitrocellulose membrane 4 and an absorption pad 2 which are sequentially arranged from one end of a pad card to the other end, wherein the nitrocellulose membrane is provided with a quality control line 5 and two detection lines 6 and 7 which are parallel to each other at intervals, and the sample pad 1 and the absorption pad 2 are respectively used for pressing two ends of the nitrocellulose membrane.
In one embodiment of the invention, preferably, a protective layer is coated outside the test strip, the waterproof layer comprises a waterproof silica gel cushion layer, a vinyl adsorption layer and a waterproof breathable paper layer which are sequentially arranged from inside to outside in a stacking manner, and the pore diameter of the breathable hole of the waterproof breathable paper layer is 10-15 μm. The outside of this protective layer sets up waterproof ventilative paper layer, can, for example contain ethylene in the air, ethylene adsorbed layer (including potassium permanganate) adsorbable ethylene so, avoids getting into inside and arouses the test paper strip rotten, sets up waterproof silica gel bed course simultaneously in inside, not only plays further waterproof effect, still has the anti effect of jolting of protection, also can completely cut off the air simultaneously, prolongs the life-span of test paper strip, extension product shelf life.
In one embodiment of the present invention, preferably, the detection substance is stored under refrigeration.
In one embodiment of the present invention, the method preferably comprises the following steps: dissolving the detection substance in the reaction buffer solution, adding the plasma or serum to be detected, mixing and adding the mixture to the test strip.
In order to ensure that the sample fully reacts with the fluorescence-labeled AMH antibody complex and the quality control complex and ensure the sensitivity of the reaction, the fluorescence-labeled AMH antibody complex, the biotin-labeled AMH antibody complex and the quality control complex are made into detection substances, the fluorescence-labeled AMH antibody complex, the biotin-labeled AMH antibody complex and the quality control complex can be dissolved only by adding corresponding solvents during the reaction, the sample is added and mixed at the moment, so that the antigen in the sample can fully react with the fluorescence-labeled AMH antibody complex, the biotin-labeled AMH antibody complex and the quality control complex, and the sensitivity of the reaction is improved. The design improves the precision and the sensitivity of the anti-mullerian hormone detection on one hand, and improves the stability of the product on the other hand because the fluorescence-labeled AMH antibody complex and the quality control complex are stored in a refrigeration way.
In order to make the technical solution of the present invention better understood by those skilled in the art, the following examples are now provided for illustration:
example 1 reaction plate preparation
Purchasing and screening high-quality monoclonal antibodies, performing a pairing experiment, and screening out capture antibodies and detection antibodies with high specificity and good affinity. The streptavidin concentration is adjusted to 1-10mg/mL and chickenIgY0.01-5mg/mL by using coating diluent respectively.
Antibody coating: preparing a nitrocellulose membrane 4 and correctly placing the nitrocellulose membrane on a film sticking device, sticking the nitrocellulose membrane 4 on a gasket, setting the spraying amount of a spraying device to be 1 mu L/cm, the spraying speed to be 9cm/s, spraying detection lines 6 and 7 and a quality control line 5 on the nitrocellulose membrane in parallel for coating, wherein the interval between the detection line and the quality control line is 2-3mm, then placing the nitrocellulose membrane in an oven, and drying the nitrocellulose membrane for 30-120 minutes at 37 ℃.
Sticking a film on the reagent strip and cutting: the sample pad 1 is correctly attached to the broad side of the pad, requiring the pad to be pressed 1-2mm against the nitrocellulose membrane 4, and the absorbent pad 2 to be attached to the narrow side of the pad, requiring the pad to be pressed above the nitrocellulose membrane 4 and 0-1mm from the outer edge of the base plate. And setting the test strip cutting width of the cutting equipment to be 3.9mm, and determining whether the cut surface is flat or not.
Assembling the reagent strip: and (3) correctly installing the test strip on the bottom shell, installing the absorption pad at the terminal of the bottom shell, closing the top shell, and placing the absorption pad on a shell pressing device for pressing firmly to obtain the reaction plate for later use.
Example 2 buffer preparation
Preparation of a reaction buffer: firstly, 100-600mM NaCl and 0.01-0.5% NaN are added into purified water30.1-3% of Tween 20, 0.01-0.5% of IGEPAL CA-630 and 0.5-3% of BSA, after the above components are completely dissolved, using phosphate buffer solution to make constant volume, filtering and storing at 2-8 deg.C.
Preparation of a detection substance:
preparing a detection substance solution: the basic buffer solution comprises 0.05-10% of BSA and 0.05-5% of Sucrose in1xPBS, and appropriate antibody complexes are respectively added into the basic buffer solution components to prepare a first detection substance containing 0.01-10 mu g/mL biotin-labeled AMH antibody complex and Eu-labeled Anti-chicken IgY antibody complex; and adding a proper amount of antibody complex into the basic buffer solution component to prepare a second detection substance containing 2-25 mug/mL Eu-labeled AMH antibody complex. The solution was stirred with a stirrer for 1 hour to mix. Taking 0.01-1 mu g of the first detection substance and 0.1-3 mu g of the second detection substance, putting the first detection substance and the second detection substance into the same bottle of test tube, mixing, then putting a proper amount of reaction buffer solution and quality control substances, and confirming whether the detection result is similar to the expected value. And (5) after the detection result is confirmed to be correct, the detection substance can be subpackaged.
Subpackaging and freeze-drying detection materials: mounting 20-80 μ L pipetting head on the detecting material racking machine, weighing test tube weight after racking (20-80 μ L), confirming whether difference between the test tube weight and the well test tube weight is consistent with racking capacity, inputting racking frequency on the detecting material racking machine after confirming that the racking capacity is consistent, and starting racking. And after the subpackaging is finished, slowly lifting the clamp of the subpackaging machine, and after the clamp is removed, separating the detection substances by using a filter screen. Transferring the obtained detection substance into a stainless steel container, confirming the temperature of a freeze dryer machine tool, and putting the detection substance into a freeze dryer for freeze drying. The probe material, which was freeze dried completely, was stored at "18-28 ℃ with a humidity < 10%".
Example 3 adding formula and manufacturing ID chip
Taking a reaction plate, a buffer solution and an AMH calibrator which are qualified by intermediate product detection, wherein each calibrator has at least 5 concentrations, each concentration is tested for at least 5 times, an average value and a concentration of an area scan of each concentration calibrator are taken as a standard curve to obtain a linear equation, the numerical value of the equation is led into special software, a blank original chip is connected to a computer, a numerical code can be led into the chip through the special software, and a chip label is pasted.
Example 4 precision test
A precision experiment was carried out using a fluorescence immunoassay analyzer (model: A2000 or A5000) which was matched with the kit of the present invention.
1. Detection materials: selecting low and high concentrations of AMH calibrator, and obtaining specific concentrations as shown in the following table
2. The test method comprises the following steps:
the kit of the present invention was subjected to precision measurement using a fluorescence immunoassay analyzer (model: A2000) equipped with the kit of the present invention, and measurement was repeated 20 times for each concentration. The operation steps of A2000 are as follows:
1) the reaction plate package was opened, and the reaction plate was removed and placed horizontally.
2) A pipette was used to remove 150. mu.L of reaction buffer and place it in a test tube of the probe substance. And immediately sampled with a pipette (50 μ L serum, plasma) and placed into a test tube of probe substance containing reaction buffer.
3) Cover the lid of buffer solution test tube, rock 10 ~ 15 times in order to ensure abundant mixing.
4) A75. mu.L sample mixture was pipetted and applied to the sample well of the reaction plate.
5) The loaded reaction plate was immediately subjected to A-chamber (35 ℃ C.) reaction for 12 minutes.
6) The reacted reaction plate is inserted into a special detection device for testing.
3. And (3) analyzing test results:
and detecting the quality control product according to a test method, and analyzing a detection result.
4. Test results
TABLE 1 AMH test results
As can be seen from Table 1, the average value, the standard deviation and the CV are calculated after the low-value and high-value quality control materials of the AMH are repeatedly measured for 20 times, and the obtained results show that the CV of the kit is 5.55 percent when the low-value sample of the AMH is measured, the CV of the high-value sample is 4.34 percent, and the CV values of the high value and the low value are both less than 10 percent, so that the kit prepared by the invention has good precision, is suitable for clinical detection and meets the differentiation requirements of different clients on different detection occasions.
Example 5 accuracy test
An accuracy test was conducted using a fluorescence immunoassay analyzer (model: A2000 or A5000) equipped with the kit of the present invention.
1. Detection materials:
the chemiluminescence method value AMH is human serum with high, medium and low concentrations of 8.09, 4.39 and 0.51ng/mL respectively.
2. The test method comprises the following steps:
the kit of the present invention was subjected to accurate detection using a fluoroimmunoassay analyzer (model: A2000) equipped with the kit of the present invention, and the measurement was repeated 3 times per concentration. The operation steps of A2000 are as follows:
1) the reaction plate package was opened, and the reaction plate was removed and placed horizontally.
2) A pipette was used to remove 150. mu.L of reaction buffer and place it in a test tube of the probe substance. And immediately sampled with a pipette (50 μ L serum, plasma) and placed into a test tube of probe substance containing reaction buffer.
3) Cover the lid of buffer solution test tube, rock 10 ~ 15 times in order to ensure abundant mixing.
4) A75. mu.L sample mixture was pipetted and applied to the sample well of the reaction plate.
5) The loaded reaction plate was immediately subjected to A-chamber (35 ℃ C.) reaction for 12 minutes.
6) The reacted reaction plate is inserted into a special detection device for testing.
3. And (3) analyzing test results:
and (4) detecting the human serum according to a test method, and analyzing a detection result.
4. And (3) test results:
TABLE 2 AMH detection of relative deviation
As can be seen from Table 2, the relative deviation is calculated after the AMH3 human serum is repeatedly measured for 3 times, and the obtained result shows that the relative deviation of the kit disclosed by the invention is-5.88% -1.61% when the AMH human serum is measured, and the relative deviation is less than 10%, so that the kit prepared by the invention is good in accuracy, suitable for clinical detection and capable of meeting the differentiation requirements of different clients on different detection occasions.
Example 6 consistency test
A fluorescent immunoassay instrument (model: A2000 or A5000) equipped with the kit of the present invention was used for the consistency test.
1. Detection materials:
the clinical samples are provided by related hospitals, and the total 100 samples with the electrochemiluminescence method comprise serum and homologous plasma, and the distribution interval of AMH content is 0.02-10 ng/mL.
2. The test method comprises the following steps:
the kit of the present invention was subjected to identity detection using a fluoroimmunoassay analyzer (model: A2000) equipped with the kit of the present invention, and the measurement was repeated 1 time. The operation steps of A2000 are as follows:
1) the reaction plate package was opened, and the reaction plate was removed and placed horizontally.
2) A pipette was used to remove 150. mu.L of reaction buffer and place it in a test tube of the probe substance. And immediately sampled with a pipette (50 μ L serum, plasma) and placed into a test tube of probe substance containing reaction buffer.
3) Cover the lid of buffer solution test tube, rock 10 ~ 15 times in order to ensure abundant mixing.
4) A75. mu.L sample mixture was pipetted and applied to the sample well of the reaction plate.
5) The loaded reaction plate was immediately subjected to A-chamber (35 ℃ C.) reaction for 12 minutes.
6) The reacted reaction plate is inserted into a special detection device for testing.
3. And (3) analyzing test results:
and (3) detecting 100 samples with the electrochemical luminescence method fixed value according to a test method, and analyzing the correlation of each sample type by taking a serum detection result as a reference.
4. And (3) test results:
as shown in FIG. 4, a scatter diagram was drawn with the serum test value of the control system as the X-axis and the serum test value of the experimental system as the Y-axis, and correlation analysis was performed. When 100 clinical serum samples are detected, the average relative deviation of the samples is less than 10%, R2 is not less than 0.98, and the slope is within the range of 0.9-1.1, so that the kit and the electrochemical luminescence method kit prepared by the invention have good consistency when used for detecting AMH serum, and meet the requirement of clinical detection of AMH serum.
As shown in FIG. 5, a scatter diagram was drawn and correlation analysis was performed with the serum measurement value of the experimental system as the X-axis and the plasma measurement value of the experimental system as the Y-axis. The detection of 100 clinical plasma samples shows that the average relative deviation of the samples is less than 10%, R2 is not less than 0.98, and the slope is within the range of 0.9-1.1, which indicates that the kit prepared by the invention has good consistency when detecting AMH serum and homologous plasma samples, and meets the requirement of clinical detection of AMH plasma.
Example 7 comparison of the Performance of the kit with the conjugate pad strip and the existing detection substance
To illustrate the differences in performance between the kits with the test substances and the strips with the binding pads, a comparative performance test study was performed.
1. Detection materials:
clinical samples are provided by relevant hospitals, 20 cases of electrochemiluminescence method-fixed value serum samples are provided, and the AMH content distribution interval is 0.02-10 ng/mL.
The AMH kit with the bonding pad and the matched instrument are purchased from related agents.
2. The test method comprises the following steps:
the kit of the present invention was subjected to identity detection using a fluoroimmunoassay analyzer (model: A2000) equipped with the kit of the present invention, and the measurement was repeated 5 times per sample. 20 serum samples were tested simultaneously using an AMH kit with conjugate pads, and each sample was tested in duplicate 5 times. The operation steps of A2000 are as follows:
1) the reaction plate package was opened, and the reaction plate was removed and placed horizontally.
2) A pipette was used to remove 150. mu.L of reaction buffer and place it in a test tube of the probe substance. And immediately sampled with a pipette (50 μ L serum, plasma) and placed into a test tube of probe substance containing reaction buffer.
3) Cover the lid of buffer solution test tube, rock 10 ~ 15 times in order to ensure abundant mixing.
4) A75. mu.L sample mixture was pipetted and applied to the sample well of the reaction plate.
5) The loaded reaction plate was immediately subjected to A-chamber (35 ℃ C.) reaction for 12 minutes.
6) The reacted reaction plate is inserted into a special detection device for testing.
3. And (3) analyzing test results:
20 cases of the electrochemiluminescence-method-fixed-value serum samples were tested according to the test method, and the differences of the 2 methods were analyzed.
4. Test results
TABLE 3 comparison of AMH test results
As shown in table 3, 20 AMH serum samples were simultaneously detected by using the strip with the bonding pad and the kit, wherein the precision CV value of each sample is equal to or greater than 10% and the relative deviation is equal to or greater than 10% when the strip with the bonding pad is used for testing the AMH serum samples, and the precision CV value of each sample is equal to or greater than 10% and the relative deviation is equal to or greater than 10% when the kit of the patent is used for testing the AMH serum samples, which indicates that the kit of the patent has good performance and meets the requirement of clinical AMH detection.
The number of modules and the processing scale described herein are intended to simplify the description of the invention. The use, modifications and variations of the anti-mullerian hormone detection kit of the present invention will be apparent to those skilled in the art.
As described above, the AMH antibody complex, biotin-labeled AMH antibody complex and quality control complex coated on the conjugate pad are made into the detection substance, so that the problems of poor AMH test precision and sensitivity are solved, and the stability of the AMH kit is improved.
While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable in various fields of endeavor to which the invention pertains, and further modifications may readily be made by those skilled in the art, it being understood that the invention is not limited to the details shown and described herein without departing from the general concept defined by the appended claims and their equivalents.
Claims (10)
1. An anti-mullerian hormone detection kit, characterized in that the kit comprises:
a probe substance containing a fluorescently labeled AMH antibody complex, a biotin labeled AMH antibody complex, and a fluorescently labeled quality control complex;
a reaction buffer stored separately from the detection substance, the reaction buffer being capable of dissolving the detection substance.
2. The anti-mullerian hormone test kit as claimed in claim 1, wherein the detection substance is a spherical particle, the detection substance comprises a first detection substance and a second detection substance, and when in use, 1 particle of each of the first detection substance and the second detection substance is dissolved in 100 to 200 μ L of the reaction buffer.
3. The anti-mullerian hormone test kit as claimed in claim 1, wherein the probe material is prepared by a process comprising:
preparing a detection substance solution: the basic buffer solution comprises 0.05-10% of BSA (bovine serum albumin) by mass percentage and 0.05-5% of sucrose by mass percentage, a first detection substance solution containing 0.01-10 mu g/mL biotin-labeled AMH antibody complex and 0.01-10 mu g/mL Eu-labeled Anti-chicken IgY antibody complex is prepared from the basic buffer solution, and then the first detection substance solution is prepared into solid particles to obtain a first detection substance; preparing a second detection substance solution containing 2-25 μ g/mL Eu-labeled AMH antibody complex by using another basic buffer solution component, and preparing the second detection substance solution into solid particles to obtain a second detection substance.
4. The anti-mullerian hormone test kit of claim 3, wherein each 1 particle of the first and second detector materials comprises 20 to 80 μ L of the active ingredient of the first and second detector material solutions, respectively.
5. The anti-mullerian hormone test kit of claim 2, wherein the mass ratio of the first probe substance to the second probe substance is from 0.01 to 1: 0.1-3.
6. The anti-mullerian hormone assay kit of claim 1, wherein the reaction buffer is prepared by dissolving the following components in 10mM phosphate buffer: 100-600mM NaCl, 0.01-0.5% (w/v) NaN3Tween 20 of 0.1-3% (v/v), IGEPAL CA-630 of 0.01-0.5% (v/v), BSA of 0.5-3% (w/v).
7. The anti-mullerian hormone test kit of claim 1, further comprising a test strip without a binding pad;
the test strip comprises a sample pad, a nitrocellulose membrane and an absorption pad which are sequentially arranged from one end of the pad card to the other end, wherein the nitrocellulose membrane is provided with a quality control line and two detection lines which are parallel to each other at intervals, and the sample pad and the absorption pad respectively press two ends of the nitrocellulose membrane.
8. The anti-mullerian hormone detection kit of claim 7, wherein the test strip is coated with a protective layer, the waterproof layer comprises a waterproof silica gel cushion layer, an ethylene adsorption layer and a waterproof breathable paper layer which are sequentially arranged from inside to outside and are stacked, and the pore diameter of the breathable hole of the waterproof breathable paper layer is 10-15 μm.
9. The anti-mullerian hormone test kit of claim 1, wherein the probe substance is stored under refrigeration.
10. A method for detecting anti-mullerian hormone using a kit as claimed in claim 1, comprising the steps of: dissolving the detection substance in the reaction buffer solution, adding the plasma or serum to be detected, mixing and adding the mixture to the test strip.
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| CN202010420025.2A CN111537752A (en) | 2020-05-18 | 2020-05-18 | Anti muller tube hormone detect reagent box |
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Cited By (1)
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| CN117405876A (en) * | 2023-11-10 | 2024-01-16 | 巴迪泰（广西）生物科技有限公司 | Three-item combined detection kit for diagnosing heart diseases and application method thereof |
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Application publication date: 20200814 |