CN106771247A - A kind of diagnostic kit of the microglobulins of β 2 - Google Patents
A kind of diagnostic kit of the microglobulins of β 2 Download PDFInfo
- Publication number
- CN106771247A CN106771247A CN201611256520.4A CN201611256520A CN106771247A CN 106771247 A CN106771247 A CN 106771247A CN 201611256520 A CN201611256520 A CN 201611256520A CN 106771247 A CN106771247 A CN 106771247A
- Authority
- CN
- China
- Prior art keywords
- reagent
- human
- latex
- diagnostic kit
- antibodies
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Abstract
The invention belongs to technical field of medical examination, and in particular to a kind of diagnostic kit of the microglobulins of β 2.The kit includes that reagent R1 and reagent R2, the reagent R1 are made up of potassium dihydrogen phosphate, lauryl glucosyl, single glycol ether of dodecyl nine, sodium chloride, magnesium chloride, calcium chloride, p-hydroxybenzoate and water；The reagent R2 is made up of the glycol ether of single dodecyl nine, lauryl glucosyl, the microglobulin antibody latex of anti-human β 2, p-hydroxybenzoate and water；A diameter of 130 150nm of the microglobulin antibody latex of the anti-human β 2.The diagnostic kit of the microglobulins of β 2 that the present invention is provided has the advantages that measure sensitivity is high, the testing result degree of accuracy is high, reproducible and stability is high, it is a kind of diagnostic kit of the ideal microglobulins of β 2, is conducive to the promotion and application of the kit.
Description
Technical field
The invention belongs to technical field of medical examination, and in particular to a kind of diagnostic kit of B2M.
Background technology
B2M (β 2-microglobin, β 2-MG), is a β light chain of the compatible antigen of tissue (HLA),
Synthesized and secreted by lymphocyte and karyocyte.The intramolecular of B2M contains a pair of disulfide bond, without sugar, its serum
Protein electrophoresis is located at the regions of β 2.Almost all of karyocyte can synthesize B2M in human body, but main by lymph
Cell synthesizes.B2M is widely present in the body fluid such as human serum, cerebrospinal fluid, urine, colostrum, saliva, and content is micro-
It is small.
Diabetic nephropathy is very common in long-term diabetic, and long-run development can more seriously damage renal function, seriously
The life quality for influenceing diabetic is the financial burden for aggravating these patients.Early diagnosis diabetic nephropathy is simultaneously carried out to it
Intervention can improve the life quality of diabetic.At present, it is urea and creatinine in clinical conventional renal function Testing index,
There is advantage simple to operate, cheap during detection and be widely used in the detection of renal function in urea and creatinine
In.But above-mentioned the two Testing index are easily by the shadow of the factors such as age, sex, musculature content and protein intake
Ring, its assessment to renal function has that the degree of accuracy is low, stablize poor defect.
The metabolism of B2M relies only on domain kidney, has cell surface to come off or be released into the B2M of blood,
Freely filtered from glomerulus, almost all of proximal tubular is received and catabolism again, no longer reflux enters blood, thus normal
In the case of serum beta-2-microglobulin can hold stabilization level.And B2M molecular weight is small, it is thus possible to from
Glomerulus is freely filtered, and catabolism of the B2M in addition to by RE rarely outside kidney, therefore, its
Generation constant rate in vivo, will not be influenceed by age, sex, body musculature and be influenceed its serum levels content.
In the case of B2M generation is not increased in vivo, it is reflection glomerular filtration work(that serum beta-2-microglobulin level is raised
Extremely sensitive index can be damaged.
Li Libo etc. has delivered the paper of entitled " development of B2M enzyme-linked immunoassay kit ",
The paper is coated with microwell plate with B2M antibody, and tetramethyl benzidine is substrate, with horseradish peroxidase-labeled β 2-
Microglobulin, establishes B2M enzyme-linked immunoassay kit.The kit is related to radioactive immunoassay
Coefficient is 0.9000, but also with simple to operate, quick advantage.But, the kit detection sensitivity is relatively low, it is impossible to full
The clinical detection requirement of the foot B2M small to content, greatly influences the diagnosis of clinical renal disease.
Patent document CN101813700B disclosed a kind of Nano microsphere immunoturbidimetry detection β on March 31st, 2010
2- microglobulin kits, the kit includes reagent R1, reagent R2 and reference calibrations product；The reagent R1 is by trihydroxy methyl
Aminomethane, wide-spectrum bactericide, disodium ethylene diamine tetraacetate, NaCl, Tween-20, PEG-6000, purified water composition；The examination
Agent R2 is made up of trishydroxymethylaminomethane, the Nano microsphere for being combined with anti-human B2M antibody, preservative, described to receive
Rice microsphere diameter is 50-150nm, and the kit for preparing is high, reproducible without dilution, the simple to operate, degree of accuracy, is applicable
In various types of automatic clinical chemistry analyzers.But, the kit sensitivity is relatively low, influences the diagnosis of clinical renal disease,
Greatly limit the application of the kit.
Therefore, research and develop out that a kind of sensitivity is high, good stability beta 2-microglobulin detecting kit is still current
The problem of urgent need to resolve.
The content of the invention
In order to overcome B2M diagnostic kit in the prior art to exist, detection sensitivity is relatively low, stability is poor
Defect, it is an object of the invention to provide a kind of diagnostic kit of B2M, to solve the above problems.
The invention provides a kind of diagnostic kit of B2M, the kit includes reagent R1 and reagent R2,
The reagent R1 is made up of following components and its concentration：
Potassium dihydrogen phosphate 120-160mmol/L, lauryl glucosyl 60-120mmol/L, single second two of dodecyl nine
Alcohol ether 100-140mmol/L, sodium chloride 3-5mmol/L, magnesium chloride 1-3mmol/L, calcium chloride 2-4mmol/L and para hydroxybenzene first
Acid esters 1-5mmol/L, balance of water；
The reagent R2 is made up of following components and its concentration：The glycol ether 100-140mmol/L of single dodecyl nine, ten
Dialkyl group glucoside 60-120mmol/L, anti-human B2M antibody latex 5-20g/L and p-hydroxybenzoate 1-
5mmol/L, balance of water；A diameter of 130-150nm of the anti-human B2M antibody latex.
Further, the kit includes reagent R1 and reagent R2, and the reagent R1 is by following components and its concentration group
Into：
Potassium dihydrogen phosphate 140mmol/L, lauryl glucosyl 80mmol/L, single glycol ether of dodecyl nine
120mmol/L, sodium chloride 4mmol/L, magnesium chloride 2mmol/L, calcium chloride 3mmol/L and p-hydroxybenzoate 2mmol/L, it is remaining
It is water to measure；
The reagent R2 is made up of following components and its concentration：The glycol ether 120mmol/L of single dodecyl nine, dodecane
Base glucoside 80mmol/L, anti-human B2M antibody latex 12g/L and p-hydroxybenzoate 2mmol/L, it is balance of
Water；A diameter of 130-150nm of the anti-human B2M antibody latex.
Further, the pH value of the reagent R1 is 7.5-7.8.
Further, the pH value of the reagent R1 is 7.6.
Further, the volume ratio of the reagent R1 and reagent R2 is 4:1.
Further, a diameter of 142nm of the anti-human B2M antibody latex.
Further, the preparation method of the anti-human B2M antibody latex is：
Anti-human B2M Antibodies Polyclonal antibodies and glue are diluted at room temperature with the kaliumphosphate buffer of 0.1mol/L
The mixture of breast, mixes on magnetic stirring apparatus, the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
In the weight ratio of anti-human B2M Antibodies Polyclonal antibodies and latex be 1:2-4, the addition of the kaliumphosphate buffer
It is 1-2 times of weight of latex；Then stirring under 37 DEG C of constant temperature is placed on, the rotating speed is 300rpm, adds and contains sweet ammonia
Closed 1 hour in the kaliumphosphate buffer of the 0.1mol/L of acid, supernatant is removed in centrifugation, add the potassium phosphate buffering of 0.1mol/L
Liquid, obtains final product.
Further, anti-human β 2- microballoons in the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
Protein antibodies polyclonal antibody is 1 with the weight ratio of latex:3.
Further, the addition of the kaliumphosphate buffer is 1.5 times of weight of latex.
The molecule of the lauryl glucosyl in the reagent R1 of the diagnostic kit of the B2M that the present invention is provided
Formula is C18H36O6, No. CAS is 110615-47-9；The molecular formula of the glycol ether of single dodecyl nine is C30H62O10, No. CAS is
3055-99-0。
Calibration object, purchases the multinomial high level immune algorithm liquid of Li De Man, and standard value is shown in specification.
Quality-control product, purchases the multinomial immune quality controlled serum of Li De Man, and Quality Control value is shown in specification.
Further, the anti-human B2M polyclonal antibody includes many grams of the anti-human B2M antibody of goat-anti
Grand antibody or the anti-human B2M polyclonal antibody of rabbit-anti or the anti-human B2M Antibodies Polyclonal antibodies of mouse, it is above-mentioned
B2M polyclonal antibody is purchased from this bio tech ltd of Beijing Ah, above-mentioned antibody only with people's β 2- microballoon eggs
White reaction, with other antigens without immunological cross-reaction, potency disclosure satisfy that this reagent requirement.Latex is purchased from
BangsLaboratories companies.
The Cleaning Principle of diagnostic kit of the B2M that the present invention is provided is：In human serum B2M with
B2M antibody in antiserum reagent forms antigen antibody complex, reaction turbidity is occurred, is detected under 600nm
The concentration for measuring B2M in serum is compared in the change of absorbance with calibration object.Specially：First add reagent R1 (samples
Dilution), electronic shell and hydrated sheath in releasing sample around antigen make antigen site fully expose；It is subsequently adding reagent R2
(anti-human B2M antibody latex solution), makes anti-human B2M antibody latex β 2- microballoons corresponding with sample
Proteantigen reacts, and forms insoluble antigen-antibody complex, produces certain turbidity, the β in its turbidity height and sample
2- microglobulin contents are directly proportional；The absorbance of the insoluble antigen-antibody complex is determined under provision wavelengths, it is and known
The B2M reference calibrations product of concentration are compared, then can calculate the content of B2M in sample.
Further, found through experiment, the diagnostic kit detection high, medium and low three of the B2M that the present invention is provided
The variation within batch coefficient of the serum sample of individual concentration value B2M is respectively 2.51%, 2.86% and 2.50%, batch anaplasia
Different coefficient is respectively 3.38%, 2.38% and 3.03%, is below 5%, illustrates the diagnosis of the B2M of present invention offer
Kit has precision higher.
Further, the detection of the diagnostic kit of the B2M that the present invention is provided is limited to 0.2mg/L, and it is 20
The R of sensitivity and linear equation after placing 6 months and 12 months DEG C respectively2Value is substantially unchanged, is preserved 36 months at 4 DEG C
Afterwards, kit sensitivity and linear good, with the firm kit for preparing without significant difference, illustrate the β 2- microballoons that the present invention is provided
The diagnostic kit of albumen has stability and preferable sensitivity higher.
In addition, present invention also offers diagnostic kit the answering in B2M is detected of the B2M
With.
In a word, compared with prior art, the diagnostic kit of the B2M that the present invention is provided has measure sensitivity
The high, advantage that the testing result degree of accuracy is high, reproducible and stability is high, is a kind of examining for ideal B2M
Disconnected kit, is conducive to the promotion and application of the kit.
Specific embodiment
The present invention is further described below by way of specific embodiment, the present invention is not limited only to following examples.In this hair
In bright scope or not departing from present disclosure, spirit and scope, the change that is carried out to the present invention, combine or replace
Change, will be apparent to the person skilled in the art, and be included within the scope of the present invention.
Embodiment 1, a kind of diagnostic kit of B2M
The diagnostic kit of the B2M includes that volume ratio is 4:1 reagent R1 and reagent R2
The reagent R1 is made up of following components and its concentration：Potassium dihydrogen phosphate 120mmol/L, lauryl glucosyl
60mmol/L, single glycol ether 100mmol/L of dodecyl nine, sodium chloride 3mmol/L, magnesium chloride 1mmol/L, calcium chloride
2mmol/L and p-hydroxybenzoate 1mmol/L, balance of water；The pH value of the reagent R1 is 7.5.
Reagent R1 preparation methods are：By above-mentioned potassium dihydrogen phosphate, lauryl glucosyl, single ethylene glycol of dodecyl nine
Ether, sodium chloride, magnesium chloride, calcium chloride and p-hydroxybenzoate add appropriate water dissolves, then add water and adjust pH to 7.5,
Obtain final product.
The reagent R2 is made up of following components and its concentration：The glycol ether 100mmol/L of single dodecyl nine, dodecane
Base glucoside 60mmol/L, anti-human B2M antibody latex 5g/L and p-hydroxybenzoate 1mmol/L, it is balance of
Water；A diameter of 130nm of the anti-human B2M antibody latex；
The preparation method of the anti-human B2M antibody latex is：
Anti-human B2M Antibodies Polyclonal antibodies and glue are diluted at room temperature with the kaliumphosphate buffer of 0.1mol/L
The mixture of breast, mixes on magnetic stirring apparatus, the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
In the weight ratio of anti-human B2M Antibodies Polyclonal antibodies and latex be 1:2, the addition of the kaliumphosphate buffer is
1 times of weight of latex；Then stirring under 37 DEG C of constant temperature is placed on, the rotating speed is 300rpm, and addition contains glycine
Closed 1 hour in the kaliumphosphate buffer of 0.1mol/L, supernatant is removed in centrifugation, adds the kaliumphosphate buffer of 0.1mol/L, i.e.,
.
Reagent R2 preparation methods are：By the above-mentioned glycol ether of single dodecyl nine, lauryl glucosyl, anti-human β 2-
Microglobulin antibody latex and p-hydroxybenzoate are added to the water and stir, and obtain final product.
Embodiment 2, a kind of diagnostic kit of B2M
The diagnostic kit of the B2M includes that volume ratio is 4:1 reagent R1 and reagent R2,
The reagent R1 is made up of following components and its concentration：Potassium dihydrogen phosphate 140mmol/L, lauryl glucosyl
80mmol/L, single glycol ether 120mmol/L of dodecyl nine, sodium chloride 4mmol/L, magnesium chloride 2mmol/L, calcium chloride
3mmol/L and p-hydroxybenzoate 2mmol/L, balance of water；The pH value of the reagent R1 is 7.6；The system of the reagent R1
Preparation Method such as embodiment 1.
The reagent R2 is made up of following components and its concentration：The glycol ether 120mmol/L of single dodecyl nine, dodecane
Base glucoside 80mmol/L, anti-human B2M antibody latex 12g/L and p-hydroxybenzoate 2mmol/L, it is balance of
Water；A diameter of 142nm of the anti-human B2M antibody latex；
The preparation method of the anti-human B2M antibody latex is：
Anti-human B2M Antibodies Polyclonal antibodies and glue are diluted at room temperature with the kaliumphosphate buffer of 0.1mol/L
The mixture of breast, mixes on magnetic stirring apparatus, the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
In the weight ratio of anti-human B2M Antibodies Polyclonal antibodies and latex be 1:3, the addition of the kaliumphosphate buffer is
1 times of weight of latex；Then stirring under 37 DEG C of constant temperature is placed on, the rotating speed is 300rpm, and addition contains glycine
Closed 1 hour in the kaliumphosphate buffer of 0.1mol/L, supernatant is removed in centrifugation, adds the kaliumphosphate buffer of 0.1mol/L, i.e.,
.
The preparation method of the reagent R1 such as embodiment 1.
Embodiment 3, a kind of diagnostic kit of B2M
The diagnostic kit of the B2M includes that volume ratio is 4:1 reagent R1 and reagent R2
The reagent R1 is made up of following components and its concentration：Potassium dihydrogen phosphate 160mmol/L, lauryl glucosyl
120mmol/L, single glycol ether 140mmol/L of dodecyl nine, sodium chloride 5mmol/L, magnesium chloride 3mmol/L, calcium chloride
4mmol/L and p-hydroxybenzoate 3mmol/L, balance of water；The pH value of the reagent R1 is 7.8；The system of the reagent R1
Preparation Method such as embodiment 1.
The reagent R2 is made up of following components and its concentration：The glycol ether 140mmol/L of single dodecyl nine, dodecane
Base glucoside 120mmol/L, anti-human B2M antibody latex 20g/L and p-hydroxybenzoate 3mmol/L, surplus
It is water；A diameter of 150nm of the anti-human B2M antibody latex；
The preparation method of the anti-human B2M antibody latex is：
Anti-human B2M Antibodies Polyclonal antibodies and glue are diluted at room temperature with the kaliumphosphate buffer of 0.1mol/L
The mixture of breast, mixes on magnetic stirring apparatus, the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
In the weight ratio of anti-human B2M Antibodies Polyclonal antibodies and latex be 1:4, the addition of the kaliumphosphate buffer is
1 times of weight of latex；Then stirring under 37 DEG C of constant temperature is placed on, the rotating speed is 300rpm, and addition contains glycine
Closed 1 hour in the kaliumphosphate buffer of 0.1mol/L, supernatant is removed in centrifugation, adds the kaliumphosphate buffer of 0.1mol/L, i.e.,
.
The preparation method of the reagent R2 such as embodiment 1.
Comparative example 1, a kind of diagnostic kit of B2M
The diagnostic kit of the B2M includes that volume ratio is 4:1 reagent R1 and reagent R2,
The reagent R1 is made up of following components and its concentration：Potassium dihydrogen phosphate 140mmol/L, Tween-20 80mmol/L,
The glycol ether 120mmol/L of single dodecyl nine, sodium chloride 4mmol/L, magnesium chloride 2mmol/L, calcium chloride 3mmol/L and to hydroxyl
Yl benzoic acid ester 2mmol/L, balance of water；The pH value of the reagent R1 is 7.6；The preparation method such as embodiment of the reagent R1
1。
The reagent R2 is made up of following components and its concentration：The glycol ether 120mmol/L of single dodecyl nine, dodecane
Base glucoside 80mmol/L, anti-human B2M antibody latex 12g/L and p-hydroxybenzoate 2mmol/L, it is balance of
Water；A diameter of 142nm of the anti-human B2M antibody latex；
The preparation method of the anti-human B2M antibody latex is similar to Example 2.
The preparation method of the reagent R2 such as embodiment 1.
Difference with embodiment 2 is：Lauryl glucosyl is replaced with into Tween-20.
Comparative example 2, a kind of diagnostic kit of B2M
The diagnostic kit of the B2M includes that volume ratio is 4:1 reagent R1 and reagent R2,
The reagent R1 is made up of following components and its concentration：Potassium dihydrogen phosphate 140mmol/L, single second two of dodecyl nine
Alcohol ether 200mmol/L, sodium chloride 4mmol/L, magnesium chloride 2mmol/L, calcium chloride 3mmol/L and p-hydroxybenzoate 2mmol/
L, balance of water；The pH value of the reagent R1 is 7.6；The preparation method of the reagent R1 such as embodiment 1.
The reagent R2 is made up of following components and its concentration：The glycol ether 120mmol/L of single dodecyl nine, dodecane
Base glucoside 80mmol/L, anti-human B2M antibody latex 12g/L and p-hydroxybenzoate 2mmol/L, it is balance of
Water；A diameter of 142nm of the anti-human B2M antibody latex；
The preparation method of the anti-human B2M antibody latex is similar to Example 2.
The preparation method of the reagent R2 such as embodiment 1.
Difference with embodiment 2 is：Without addition lauryl glucosyl, increase single glycol ether of dodecyl nine
Consumption.
Comparative example 3, a kind of diagnostic kit of B2M
The diagnostic kit of the B2M includes that volume ratio is 4:1 reagent R1 and reagent R2,
The reagent R1 is made up of following components and its concentration：Potassium dihydrogen phosphate 140mmol/L, lauryl glucosyl
80mmol/L, single glycol ether 120mmol/L of dodecyl nine, sodium chloride 4mmol/L, magnesium chloride 2mmol/L, calcium chloride
3mmol/L and p-hydroxybenzoate 2mmol/L, balance of water；The pH value of the reagent R1 is 7.6；The system of the reagent R1
Preparation Method such as embodiment 1.
The reagent R2 is made up of following components and its concentration：The glycol ether 120mmol/L of single dodecyl nine, dodecane
Base glucoside 80mmol/L, anti-human B2M antibody latex 12g/L and p-hydroxybenzoate 2mmol/L, it is balance of
Water；A diameter of 125nm of the anti-human B2M antibody latex；
The preparation method of the anti-human B2M antibody latex is similar to Example 2.
The preparation method of the reagent R2 such as embodiment 1.
Difference with embodiment 2 is：A diameter of 125nm of the anti-human B2M antibody latex.
Comparative example 4, a kind of diagnostic kit of B2M
The diagnostic kit of the B2M includes that volume ratio is 4:1 reagent R1 and reagent R2,
The component and concentration of the reagent R1 and reagent R2 such as embodiment 2.
The preparation method of the reagent R1 and reagent R2 such as embodiment 1.
The preparation method of the anti-human B2M antibody latex is：
Anti-human B2M Antibodies Polyclonal antibodies and glue are diluted at room temperature with the kaliumphosphate buffer of 0.1mol/L
The mixture of breast, mixes on magnetic stirring apparatus, the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
In the weight ratio of anti-human B2M Antibodies Polyclonal antibodies and latex be 1:1, the addition of the kaliumphosphate buffer is
1 times of weight of latex；Then stirring under 37 DEG C of constant temperature is placed on, the rotating speed is 300rpm, and addition contains glycine
Closed 1 hour in the kaliumphosphate buffer of 0.1mol/L, supernatant is removed in centrifugation, adds the kaliumphosphate buffer of 0.1mol/L, i.e.,
.
Difference with embodiment 2 is：In the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
Anti-human B2M Antibodies Polyclonal antibodies are 1 with the weight ratio of latex:1.
The precision test of the diagnostic kit of test example one, B2M
1st, test material：The diagnostic kit of B2M prepared by embodiment 2, the β of high, medium and low three kinds of concentration values
The serum sample of 2- microglobulins.
2nd, test method：The method that research institute (CLSI) EP5A2 files are talked about with reference to American National Clinical Laboratory Standard, adopts
The diagnostic kit of the B2M prepared with embodiment 2 to high (4.65mg/L), in (2.45mg/L), low (0.65mg/
L) three serum samples of the B2M of concentration of value carry out repeated measure respectively, by high, medium and low three concentration value β 2-
The serum sample of microglobulin is distributed into 20 pipes respectively, is preserved in placing -20 DEG C, automatically raw using the 7600 of Japanese HITACH
Change analyzer and survey a pipe daily, METHOD FOR CONTINUOUS DETERMINATION 20 days calculates the within-run and between-run analysis coefficient (CV%) of each concentration.
3rd, result of the test：
Result of the test is as shown in table 1.
The Precision Analyze of the high, normal, basic three kinds of horizontal B2Ms of the serum of table 1
As shown in Table 1, the diagnostic kit of the B2M that the present invention is provided detects high, medium and low three concentration of value
The variation within batch coefficient of the serum sample of B2M is respectively 2.51%, 2.86% and 2.50%, interassay coefficient of variation point
Not Wei 3.38%, 2.38% and 3.03%, be below 5%, illustrate the present invention offer B2M diagnostic kit tool
There is precision higher.
The sensitivity test of the diagnostic kit of test example two, B2M
1st, test material：The diagnostic kit of B2M prepared by embodiment 2, the multinomial high level of Li De Man
Immune algorithm liquid.
2nd, test method：With reference to the test limit criterion that Japanese JJCLA files specify, by B2M standard items
Accurate dilutions are respectively determined 10 times, with measurement result into 5 low concentration samples of 0.1-0.5mg/LWith matrix liquidNonoverlapping minimum is test limit.
3rd, result of the test
Result of the test is as shown in table 2.
The sensitivity test of the diagnostic kit of the B2M of table 2
Note：* it is absorbance × 1000
As shown in Table 2, measurement result of the diagnostic kit of the B2M that the present invention is provided in 0.2mg/L concentrationWith matrix liquidDo not overlap, therefore, the detection of the diagnostic kit of the B2M of present invention offer is provided
0.2mg/L is limited to, has the advantages that detection limit is low, sensitivity is high.
The stability test of the diagnostic kit of test example three, B2M
1st, test material：Embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3 and comparative example 4 are made
The diagnostic kit of standby B2M.
2nd, test method：
β 2- prepared by embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3 and comparative example 4 are micro-
After the diagnostic kit of globulin is placed 6 months and 12 months respectively at 20 DEG C, kit is determined according to the method for test example two
Sensitivity, and β prepared by embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3 and comparative example 4
The diagnostic kit of 2- microglobulins detects serum beta-2-microglobulin respectively with Li Deman reagents, then carries out the above results
Regression analysis, calculates R2Value.
3rd, result of the test：
Result of the test is as shown in table 3.
The stability test of the diagnostic kit of the B2M of table 3
As shown in Table 3, the diagnostic kit of the B2M that the present invention is provided places 6 months and 12 respectively at 20 DEG C
The R of sensitivity and linear equation after individual month2Value is substantially unchanged, illustrates the diagnostic reagent of the B2M of present invention offer
Box has stability higher and preferable sensitivity.
In addition, after the diagnostic kit of the B2M of present invention offer is preserved 36 months at 4 DEG C, kit spirit
Sensitivity and linear good, with the firm kit for preparing without significant difference.
Claims (9)
1. a kind of diagnostic kit of B2M, it is characterised in that the kit includes reagent R1 and reagent R2, institute
Reagent R1 is stated to be made up of following components and its concentration：
Potassium dihydrogen phosphate 120-160mmol/L, lauryl glucosyl 60-120mmol/L, single glycol ether of dodecyl nine
100-140mmol/L, sodium chloride 3-5mmol/L, magnesium chloride 1-3mmol/L, calcium chloride 2-4mmol/L and p-hydroxybenzoate
1-5mmol/L, balance of water；
The reagent R2 is made up of following components and its concentration：
The glycol ether 100-140mmol/L of single dodecyl nine, lauryl glucosyl 60-120mmol/L, anti-human β 2- are micro-
Globulin antibody latex 5-20g/L and p-hydroxybenzoate 1-5mmol/L, balance of water；The anti-human B2M resists
A diameter of 130-150nm of body latex.
2. the diagnostic kit of B2M as claimed in claim 1, it is characterised in that the kit includes reagent
R1 and reagent R2, the reagent R1 are made up of following components and its concentration：
Potassium dihydrogen phosphate 140mmol/L, lauryl glucosyl 80mmol/L, single glycol ether 120mmol/ of dodecyl nine
L, sodium chloride 4mmol/L, magnesium chloride 2mmol/L, calcium chloride 3mmol/L and p-hydroxybenzoate 2mmol/L, balance of water；
The reagent R2 is made up of following components and its concentration：
The glycol ether 120mmol/L of single dodecyl nine, lauryl glucosyl 80mmol/L, anti-human B2M resist
Body latex 12g/L and p-hydroxybenzoate 2mmol/L, balance of water；The diameter of the anti-human B2M antibody latex
It is 130-150nm.
3. the diagnostic kit of B2M as claimed in claim 1 or 2, it is characterised in that the pH value of the reagent R1
It is 7.5-7.8.
4. the diagnostic kit of B2M as claimed in claim 3, it is characterised in that the pH value of the reagent R1 is
7.6。
5. the diagnostic kit of B2M as claimed in claim 1 or 2, it is characterised in that the reagent R1 and reagent
The volume ratio of R2 is 4:1.
6. the diagnostic kit of B2M as claimed in claim 1 or 2, it is characterised in that the anti-human β 2- microballoons
A diameter of 142nm of protein antibodies latex.
7. the diagnostic kit of B2M as claimed in claim 1 or 2, it is characterised in that the anti-human β 2- microballoons
The preparation method of protein antibodies latex is：
Anti-human B2M Antibodies Polyclonal antibodies and latex are diluted at room temperature with the kaliumphosphate buffer of 0.1mol/L
Mixture, mixes on magnetic stirring apparatus, resists in the mixture of the anti-human B2M Antibodies Polyclonal antibodies and latex
People's B2M Antibodies Polyclonal antibodies are 1 with the weight ratio of latex:2-4, the addition of the kaliumphosphate buffer is glue
1-2 times of newborn weight；Then stirring under 37 DEG C of constant temperature is placed on, the rotating speed is 300rpm, and addition contains glycine
Closed 1 hour in the kaliumphosphate buffer of 0.1mol/L, supernatant is removed in centrifugation, adds the kaliumphosphate buffer of 0.1mol/L, i.e.,
.
8. the diagnostic kit of B2M as claimed in claim 7, it is characterised in that the anti-human B2M
Anti-human B2M Antibodies Polyclonal antibodies and the weight ratio of latex are 1 in the mixture of Antibodies Polyclonal antibodies and latex:
3。
9. the diagnostic kit of B2M as claimed in claim 7, it is characterised in that the kaliumphosphate buffer
Addition is 1.5 times of weight of latex.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611256520.4A CN106771247A (en) | 2016-12-30 | 2016-12-30 | A kind of diagnostic kit of the microglobulins of β 2 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611256520.4A CN106771247A (en) | 2016-12-30 | 2016-12-30 | A kind of diagnostic kit of the microglobulins of β 2 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106771247A true CN106771247A (en) | 2017-05-31 |
Family
ID=58954151
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611256520.4A Pending CN106771247A (en) | 2016-12-30 | 2016-12-30 | A kind of diagnostic kit of the microglobulins of β 2 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106771247A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107543931A (en) * | 2017-09-12 | 2018-01-05 | 四川新健康成生物股份有限公司 | Kit based on latex enhancing immune turbidimetry detection TnI and preparation method thereof |
| CN109187984A (en) * | 2018-08-11 | 2019-01-11 | 金华市强盛生物科技有限公司 | A kind of β2-microglobulin detection kit |
| CN111239403A (en) * | 2019-02-13 | 2020-06-05 | 武汉生之源生物科技股份有限公司 | β 2 microglobulin latex enhanced immunoturbidimetry kit and application thereof |
| CN111579772A (en) * | 2020-05-28 | 2020-08-25 | 珠海丽珠试剂股份有限公司 | Detection reagent based on immunomagnetic bead method, preparation method thereof, kit and detection method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101813700A (en) * | 2010-03-31 | 2010-08-25 | 浙江伊利康生物技术有限公司 | Kit for detecting beta2-microglobulin by nanometer microsphere immunoturbidimetry |
| CN102590526A (en) * | 2012-01-13 | 2012-07-18 | 宁波美康生物科技股份有限公司 | Beta 2-microglobulin detection kit |
| CN103995129A (en) * | 2014-05-08 | 2014-08-20 | 北京玖佳宜科技有限公司 | Beta 2-microglobulin detection kit and preparation |
| CN105102629A (en) * | 2013-01-22 | 2015-11-25 | 艾米克罗齐公司 | Rapid method for detection of pathogen |
| CN106053840A (en) * | 2016-07-12 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | Kit for measuring alpha1-microglobulin and preparation method of kit |
-
2016
- 2016-12-30 CN CN201611256520.4A patent/CN106771247A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101813700A (en) * | 2010-03-31 | 2010-08-25 | 浙江伊利康生物技术有限公司 | Kit for detecting beta2-microglobulin by nanometer microsphere immunoturbidimetry |
| CN102590526A (en) * | 2012-01-13 | 2012-07-18 | 宁波美康生物科技股份有限公司 | Beta 2-microglobulin detection kit |
| CN105102629A (en) * | 2013-01-22 | 2015-11-25 | 艾米克罗齐公司 | Rapid method for detection of pathogen |
| CN103995129A (en) * | 2014-05-08 | 2014-08-20 | 北京玖佳宜科技有限公司 | Beta 2-microglobulin detection kit and preparation |
| CN106053840A (en) * | 2016-07-12 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | Kit for measuring alpha1-microglobulin and preparation method of kit |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107543931A (en) * | 2017-09-12 | 2018-01-05 | 四川新健康成生物股份有限公司 | Kit based on latex enhancing immune turbidimetry detection TnI and preparation method thereof |
| CN107543931B (en) * | 2017-09-12 | 2020-02-21 | 四川新健康成生物股份有限公司 | Kit for detecting TnI based on latex enhanced immunoturbidimetry and preparation method thereof |
| CN109187984A (en) * | 2018-08-11 | 2019-01-11 | 金华市强盛生物科技有限公司 | A kind of β2-microglobulin detection kit |
| CN111239403A (en) * | 2019-02-13 | 2020-06-05 | 武汉生之源生物科技股份有限公司 | β 2 microglobulin latex enhanced immunoturbidimetry kit and application thereof |
| CN111239403B (en) * | 2019-02-13 | 2020-12-08 | 武汉生之源生物科技股份有限公司 | Beta 2 microglobulin latex enhanced immunoturbidimetry kit and application |
| CN111579772A (en) * | 2020-05-28 | 2020-08-25 | 珠海丽珠试剂股份有限公司 | Detection reagent based on immunomagnetic bead method, preparation method thereof, kit and detection method |
| CN111579772B (en) * | 2020-05-28 | 2022-04-08 | 珠海丽珠试剂股份有限公司 | Detection reagent based on immunomagnetic bead method, preparation method thereof, kit and detection method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102759631B (en) | The latex enhancing immune of a kind of quantitative detection Procalcitonin PCT is than turbid kit | |
| Punshon et al. | Placental metal concentrations in relation to maternal and infant toenails in a US cohort | |
| CN106771247A (en) | A kind of diagnostic kit of the microglobulins of β 2 | |
| CN101699287B (en) | Homogeneous phase sol particle type cystatin C measuring kit and preparation method thereof | |
| CN101454670B (en) | Method for measuring human megalin | |
| CN102662061A (en) | Kit for determination of human alpha-fetoprotein content by latex-enhanced immunoturbidimetry | |
| CN106568978A (en) | Serum amyloid protein A detection method and reagent | |
| CN103364558A (en) | Human tumor marker carcinoembryonic antigen (CEA) magnetic particle chemiluminiscence immunoassay kit and detection method | |
| CN111239421A (en) | Latex-enhanced immunoturbidimetry kit for quantitatively detecting adiponectin ADPN and preparation and application methods thereof | |
| CN106918708A (en) | A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin | |
| CN107860929A (en) | The immunoturbidimetry detection reagent and method of a kind of serum amyloid A protein | |
| CN106370860A (en) | Kit and test paper strip for serum immunoglobulin E colloidal gold chromatography quantitative detection | |
| CN105911298A (en) | Kit for determining myoglobin | |
| CN114200132A (en) | Kit for detecting thyroglobulin antibody and subtype thereof | |
| CN104711279A (en) | Vascular endothelium growth factor detection kit and raw material preparation | |
| CN106018299A (en) | Kit for measuring serum amyloid A and preparation method of kit | |
| Kocyigit et al. | The association of endothelin-1 levels with renal survival in polycystic kidney disease patients | |
| CN106053862A (en) | Serum amyloid protein A (SAA) measuring kit | |
| CN105699665A (en) | Detection kit of lipocalin related to neutrophil gelatinase | |
| CN106018816A (en) | Kit for measuring procalcitonin | |
| CN102890083B (en) | Chemical luminous substrate solution and containing its test kit and the detection method applying it | |
| CN102435755A (en) | Quantitative determination kit for total triiodothyronine (TT3) and detection method thereof | |
| CN102426249A (en) | Kit for quantitatively determining thyroid stimulating hormone (TSH) and detection method for TSH | |
| Cai et al. | Reference intervals of thyroid hormones in a previously iodine-deficient but presently more than adequate area of Western China: a population-based survey | |
| Offermann et al. | Development and validation of a lateral flow assay (LFA) for the determination of IgG-antibodies to Pr3 (cANCA) and MPO (pANCA) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170531 |
|
| RJ01 | Rejection of invention patent application after publication |