CN105738300B - Detect kit, method and the purposes of KAPPA light chain content - Google Patents
Detect kit, method and the purposes of KAPPA light chain content Download PDFInfo
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Abstract
The present invention discloses a kind of kit for detecting KAPPA light chain content, comprising: reagent R1, reagent R2 and KAPPA light chain calibration object；The reagent R1 includes: the first buffer, the first electrolyte, surfactant, the first stabilizer, macromolecule promotor and the first preservative；The reagent R2 includes: the second buffer, anti-human KAPPA light chain antibody, the second electrolyte, the second stabilizer and the second preservative；The C1 inhibitor calibration object includes: third buffer, third stabilizer, third preservative, antioxidant and KAPPA Light Chain Antigen.Invention additionally discloses the purposes and a kind of method of the kit detection KAPPA light chain content using detection KAPPA light chain content as described above in detection KAPPA light chain content of a kind of kit of detection KAPPA light chain content as described above.Kit and detection method of the invention succinctly can quickly detect KAPPA light chain content.
Description
Technical field
The invention belongs to detect the technical field of KAPPA light chain content, and in particular to a kind of detection KAPPA light chain content
Kit, method and purposes.
Background technique
Light chain (Light chain, L) is about made of 214 amino acid residues, is typically free of carbohydrate, molecule
Amount is about 24kD.Every light chain is containing there are two the cyclic peptide as composed by intrachain disulfide bond.L chain shares two types: KAPPA (κ)
With Lambda (λ), the type of L chain is always identical on the same native immunoglobulin molecule.In normal human serum κ: λ about
For 2:1.
The paraplasm of cell clone will lead to monoclonal globulin or intracellular immunoglobulin segment (free light chain)
Increase, this will cause the proportional imbalance of κ Yu λ chain, and the ratio of κ and λ are not normal predictive of monoclonal Y globulinemia.It is generating
In increased situation, the filtration of free light chain also increases, and when exceeding the reabsorption ability of renal tubule, which can quantitatively be examined
Survey the light chain in combination and intracellular immunoglobulin.
The different types of light chain content of quantitative detection helps to diagnose macroglobulinemia (huge globulin generation increases)
And connective tissue disease (such as rheumatoid arthritis, systemic loupus erythematosus) disease.
In addition, light chain can also increase in the blood such as infection, acute, chronic hepatitis, cirrhosis, but generally shows as κ, λ while increasing
It is more；The Urinaries such as nephrosis, diabetes also may occur in which κ, λ while increase.
Existing detection method is mainly immunoelectrophoresis, enzyme-linked immunization.The former detection time is longer, operates more numerous
It is trivial, atypical patients cannot be made and clarified a diagnosis.Enzyme-linked immunization has the expansion effect of enzymatic, quantitative not accurate enough,
Limit its application.Minute is not only greatly shortened using immunoturbidimetry, the sensibility of immunology diagnosis also can be improved
And accuracy, potential applicability in clinical practice are wide.
Summary of the invention
The above-mentioned deficiency for aiming to overcome that the prior art of the embodiment of the present invention provides a kind of detection KAPPA light chain and contains
The kit of amount succinctly can rapidly detect the content of KAPPA light chain in sample, accuracy with higher and well spy
It is anisotropic.
The another object of the embodiment of the present invention is to overcome the above-mentioned deficiency of the prior art, provides a kind of above-mentioned detection
The purposes in detection KAPPA light chain content of the kit of KAPPA light chain content can be used in succinct rapidly detection sample
The content of KAPPA light chain, accuracy with higher and well specificity.
A further object of the embodiment of the present invention is to overcome the above-mentioned deficiency of the prior art, provide a kind of using above-mentioned inspection
The method for surveying the kit detection KAPPA light chain content of KAPPA light chain content, succinctly can rapidly detect KAPPA in sample
The content of light chain, accuracy with higher and well specificity.
In order to achieve the above-mentioned object of the invention, the technical solution of the embodiment of the present invention is as follows:
A kind of kit detecting KAPPA light chain content, comprising: reagent R1, reagent R2 and KAPPA light chain calibration object；Institute
Stating reagent R1 includes: first 10~600mmol/L of buffer, first 5~100g/L of electrolyte, 0.5~60g/ of surfactant
L, first 0.5~20g/L of stabilizer, 1~100g/L of macromolecule promotor and first 1~10g/L of preservative；The reagent R2 packet
It includes: second 10~600mmol/L of buffer, 5~60%w/v of anti-human KAPPA light chain antibody, second 5~50g/L of electrolyte,
Two 0.5~10g/L of stabilizer and second 1~10g/L of preservative；The KAPPA light chain calibration object include: third buffer 10~
600mmol/L, 0.5~60g/L of third stabilizer, 1~10g/L of third preservative, antioxidant 0.01~10g/L and KAPPA
Light Chain Antigen.
Further, the pH of the reagent R1 is 6~9.5.
Further, the pH of the reagent R2 is 6~9.5.
Further, first buffer, second buffer and the third buffer are selected from acetate salt buffer
Liquid, ammonium chloride buffer, phosphate buffer, trishydroxymethylaminomethane TRIS buffer, borax-sodium hydrate buffer solution,
Glycine buffer, 3- (N- morpholine) propane sulfonic acid MOPS buffer, 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffering
One or more of liquid, 4- hydroxyethyl piperazineethanesulfonic acid HEPES buffer solution；And/or first stabilizer, described second
Stabilizer and the third stabilizer are selected from iminodiacetic acid, sodium sulfate of polyethenoxy ether of fatty alcohol, ethylenediamine tetra-acetic acid
Disodium, diethylene triamine pentacetic acid (DTPA) sodium, 1,2- cyclohexanediol glycidol, mannose, glucose, chitosan, sorbierite and ox
One or more of haemocyanin；And/or first preservative, second preservative and the third preservative are equal
Selected from one of Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate and ethyl mercury sodium thiosulfate or several
Kind；And/or first electrolyte and second electrolyte are selected from sodium chloride, magnesium sulfate, magnesium chloride and potassium chloride
It is one or more of；And/or the surfactant be selected from Triton series of surfactants, Tween series of surfactants,
One or more of laurel ether, polyoxyethylene alkyl phenyl ether, ethylene nonyl phenyl ether and polyoxethylene octylphenyl phenylate；
And/or the macromolecule promotor be selected from polyethylene glycol PEG 8000, polyethylene glycol PEG 6000, polyethylene glycol PEG 4000,
Polyethylene glycol PEG 2000, fatty alcohol polyoxyethylene ether, polyvinylpyrrolidone, Sodium Polyacrylate, ethylene polyethenoxy ether,
One or more of hydroxypropyl methyl cellulose and sodium cellulose glycolate；And/or the anti-human KAPPA light chain antibody choosing
, rabbit-anti people anti-human from mouse, goat-anti people, chicken be anti-human or the anti-human KAPPA light chain antibody of ox；The antioxidant is selected from butylhydroxy fennel
Fragrant ether, propylgallate, dibutyl hydroxy toluene, benzene polyphenol, phosphatide, Licorice root antioxidant, dilauryl thiodipropionate,
One or more of tert-butylhydroquinone and vitamin series.
Further, first buffer, second buffer and the third buffer are 3- (cyclohexylamine) -2-
Hydroxyl -1- propane sulfonic acid CAPSO buffer；And/or first stabilizer, second stabilizer and the third stabilizer
It is 1,2- cyclohexanediol glycidol；And/or first preservative, second preservative and the third preservative
It is ethyl-para-hydroxybenzoate；And/or first electrolyte and second electrolyte are sodium chloride；And/or institute
Stating surfactant is polyoxethylene octylphenyl phenylate；And/or the macromolecule promotor is hydroxypropyl methyl cellulose；With/
Or, the anti-human KAPPA light chain antibody is goat-anti people KAPPA light chain monoclonal antibody；And/or the antioxidant is two fourths
Base hydroxy-methylbenzene.
Further, the reagent R1 include: 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffer 10~
600mmol/L, 5~100g/L of sodium chloride, 0.5~60g/L of polyoxethylene octylphenyl phenylate, 1,2- cyclohexanediol glycidol 0.5
The pH of~20g/L, 1~100g/L of hydroxypropyl methyl cellulose and ethyl-para-hydroxybenzoate 1~10g/L, the reagent R1 are 6
~9.5；The reagent R2 include: 10~600mmol/L of ammonium chloride buffer, 10~60%w/v of anti-human KAPPA light chain antibody,
5~50g/L of magnesium sulfate, 0.5~10g/L of sodium sulfate of polyethenoxy ether of fatty alcohol and phenol 1~10g/L, the pH of the reagent R2
It is 6~9.5；The KAPPA light chain calibration object includes: 10~600mmol/L of glycine buffer, 5~50g/L of chitosan, right
1~10g/L of hydroxybenzoic acid, Licorice root antioxidant 0.01~10g/L and KAPPA light chain.
A kind of purposes in detection KAPPA light chain content of the kit of detection KAPPA light chain content as described above.
A method of KAPPA light chain content, packet are detected using the kit of detection KAPPA light chain content as described above
It includes: being 1~10:200 by the volume ratio of sample to be tested and the reagent R1, the reagent R1 is added in Xiang Suoshu sample to be tested,
The first mixed liquor is obtained, and obtains the absorbance OD1 of first mixed liquor；By the reagent R2 and the reagent R1 volume ratio
For 1:1~10, the reagent R2 is added in the first mixed liquor of Xiang Suoshu, obtains the second mixed liquor, and obtains second mixing
The absorbance OD2 of liquid；Standard curve is obtained using the KAPPA light chain calibration object, according to the absorbance OD1 and the extinction
Degree OD2 obtains the content of KAPPA light chain in the sample to be tested on the standard curve.
Further, the reagent R2 and the reagent R1 volume ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,
1:9 or 1:10.
Further, the fitting mode of the standard curve includes: Spline, quadratic equation, logit-log3p, logit-
Log4p, logit-log5p or Polygonal.
The embodiment of the present invention has the beneficial effect that:
1, the kit of the detection KAPPA light chain content of the embodiment of the present invention quickly using simplicity succinctly can be examined rapidly
The content of KAPPA light chain in test sample sheet.
2, the kit of the detection KAPPA light chain content of the embodiment of the present invention can satisfy clinical fast high-flux detection
The requirement of sample, detection efficiency significantly improve, and have broad prospects in clinical application.
3, the kit of the detection KAPPA light chain content of the embodiment of the present invention, also has high degree of automation, detection range
The advantages that wide.
4, the kit of the detection KAPPA light chain content of the embodiment of the present invention makes it have detection KAPPA light chain content
Purposes.
5, the method for the kit detection KAPPA light chain content of the detection KAPPA light chain content of the embodiment of the present invention, succinctly
The content of KAPPA light chain in sample is rapidly detected, detection efficiency is higher, has broad prospects in clinical application.
Detailed description of the invention
Fig. 1 is the flow chart of the method for detection KAPPA light chain content of the invention；
Fig. 2 is the flow diagram of the detection method of the KAPPA light chain content of the embodiment of the present invention 1；
Fig. 3 is the comparison result of detection the KAPPA light chain content kit and Correlation to That Abroad kit of the embodiment of the present invention 6
Schematic diagram.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, right below in conjunction with drawings and examples
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.
The present invention provides a kind of kits for detecting KAPPA light chain content.The kit includes: reagent R1, reagent R2
With KAPPA light chain calibration object.
Wherein, reagent R1 includes: first 10~600mmol/L of buffer, first 5~100g/L of electrolyte, surface-active
0.5~60g/L of agent, first 0.5~20g/L of stabilizer, 1~100g/L of macromolecule promotor and first 1~10g/L of preservative.
Reagent R2 includes: second 10~600mmol/L of buffer, 5~60%w/v of anti-human KAPPA light chain antibody, the second electricity
Solve 5~50g/L of matter, second 0.5~10g/L of stabilizer and second 1~10g/L of preservative.In the present invention, the meaning of w/v expression
It is Solute mass contained in the solution of unit volume.
KAPPA light chain calibration object includes: 10~600mmol/L of third buffer, 0.5~60g/L of third stabilizer, third
1~10g/L of preservative, antioxidant 0.01~10g/L and KAPPA Light Chain Antigen.
Preferably, reagent R1 includes: first 20~400mmol/L of buffer, first 5~30g/L of electrolyte, surface-active
1~50g/L of agent, first 0.5~5g/L of stabilizer, 10~100g/L of macromolecule promotor and first 2~10g/L of preservative.
Preferably, reagent R2 includes: second 10~400mmol/L of buffer, 16~50%w/ of anti-human KAPPA light chain antibody
V, second 6~50g/L of electrolyte, second 0.5~10g/L of stabilizer and second 3~10g/L of preservative.
Preferably, KAPPA light chain calibration object include: 10~600mmol/L of third buffer, third stabilizer 0.5~
60g/L, 1~10g/L of third preservative, antioxidant 0.01~10g/L and KAPPA Light Chain Antigen.
Preferably, the pH of reagent R1 is 6~9.5.
Preferably, the pH of reagent R2 is 6~9.5.
Preferably, in reagent R1, the first buffer be selected from acetate buffer, ammonium chloride buffer, phosphate buffer,
Trishydroxymethylaminomethane TRIS buffer, borax-sodium hydrate buffer solution, glycine buffer, 3- (N- morpholine) propane sulfonic acid
MOPS buffer, 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffer, 4- hydroxyethyl piperazineethanesulfonic acid HEPES buffering
One or more of liquid.First electrolyte is selected from one or more of sodium chloride, magnesium sulfate, magnesium chloride and potassium chloride.Table
Face activating agent is selected from Triton series of surfactants, Tween series of surfactants, laurel ether, polyoxyethylene alkylphenyl
One or more of ether, ethylene nonyl phenyl ether and polyoxethylene octylphenyl phenylate.First stabilizer is selected from imino-diacetic
Acetic acid, sodium sulfate of polyethenoxy ether of fatty alcohol, disodium ethylene diamine tetraacetate, diethylene triamine pentacetic acid (DTPA) sodium, 1,2- cyclohexanediol
One or more of glycidol, mannose, glucose, chitosan, sorbierite and bovine serum albumin.The choosing of macromolecule promotor
From polyethylene glycol PEG 8000, polyethylene glycol PEG 6000, polyethylene glycol PEG 4000, polyethylene glycol PEG 2000, fatty alcohol
Polyoxyethylene ether, polyvinylpyrrolidone, Sodium Polyacrylate, ethylene polyethenoxy ether, hydroxypropyl methyl cellulose and hydroxyl first
One or more of base sodium cellulosate.First preservative is selected from Sodium azide, phenol, P-hydroxybenzoic acid, P-hydroxybenzoic acid
One or more of ethyl ester and ethyl mercury sodium thiosulfate.
Preferably, in reagent R2, the second buffer be selected from acetate buffer, ammonium chloride buffer, phosphate buffer,
Trishydroxymethylaminomethane TRIS buffer, borax-sodium hydrate buffer solution, glycine buffer, 3- (N- morpholine) propane sulfonic acid
MOPS buffer, 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffer, 4- hydroxyethyl piperazineethanesulfonic acid HEPES buffering
One or more of liquid.Anti-human KAPPA light chain antibody is selected from mouse is anti-human, rabbit-anti people, goat-anti people, chicken is anti-human or ox is anti-human
KAPPA light chain antibody.Second electrolyte is selected from one or more of sodium chloride, magnesium sulfate, magnesium chloride and potassium chloride.Second is steady
Determine agent and is selected from iminodiacetic acid, sodium sulfate of polyethenoxy ether of fatty alcohol, disodium ethylene diamine tetraacetate, diethylene triamine pentacetic acid (DTPA)
One of sodium, 1,2- cyclohexanediol glycidol, mannose, glucose, chitosan, sorbierite and bovine serum albumin are several
Kind.Second preservative is selected from Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate and ethyl mercury sodium thiosulfate
One or more of.
Preferably, in KAPPA light chain calibration object, third buffer is selected from acetate buffer, ammonium chloride buffer, phosphoric acid
Salt buffer, trishydroxymethylaminomethane TRIS buffer, borax-sodium hydrate buffer solution, glycine buffer, 3- (N-
Quinoline) propane sulfonic acid MOPS buffer, 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffer, 4- hydroxyethyl piperazineethanesulfonic acid
One or more of HEPES buffer solution.Third stabilizer be selected from iminodiacetic acid, sodium sulfate of polyethenoxy ether of fatty alcohol,
Disodium ethylene diamine tetraacetate, diethylene triamine pentacetic acid (DTPA) sodium, 1,2- cyclohexanediol glycidol, mannose, glucose, shell are poly-
One or more of sugar, sorbierite and bovine serum albumin.Third preservative is selected from Sodium azide, phenol, P-hydroxybenzoic acid, right
One or more of nipagin A and ethyl mercury sodium thiosulfate.Antioxidant is selected from butylated hydroxy anisole, does not have
Propyl galate, dibutyl hydroxy toluene, benzene polyphenol, phosphatide, Licorice root antioxidant, dilauryl thiodipropionate, tertiary butyl
One or more of hydroquinone and vitamin series.
It is furthermore preferred that the first buffer is 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffer in reagent R1.
First electrolyte is sodium chloride.Surfactant is polyoxethylene octylphenyl phenylate.First stabilizer is the shrink of 1,2- cyclohexanediol
Glycerol.Macromolecule promotor is hydroxypropyl methyl cellulose.First preservative is ethyl-para-hydroxybenzoate.
It is furthermore preferred that the second buffer is 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffer in reagent R2.
Anti-human KAPPA light chain antibody is goat-anti people KAPPA light chain monoclonal antibody.Second electrolyte is sodium chloride.Second stabilizer is
1,2- cyclohexanediol glycidol.Second preservative is ethyl-para-hydroxybenzoate.
It is furthermore preferred that third buffer is 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid in KAPPA light chain calibration object
CAPSO buffer.Third stabilizer is 1,2- cyclohexanediol glycidol.Third preservative is ethyl-para-hydroxybenzoate.It is anti-
Oxidant is dibutyl hydroxy toluene.
It is furthermore preferred that the kit includes following ingredient:
Reagent R1 includes: 10~600mmol/L of 3- (cyclohexylamine) -2- hydroxyl -1- propane sulfonic acid CAPSO buffer, sodium chloride 5
~100g/L, 0.5~60g/L of polyoxethylene octylphenyl phenylate, 0.5~20g/L of 1,2- cyclohexanediol glycidol, hydroxypropyl methyl
1~10g/L of 1~100g/L of cellulose and ethyl-para-hydroxybenzoate.The pH of reagent R1 is 6~9.5.Reagent R2 includes: chlorination
10~600mmol/L of ammonium buffer, 10~60%w/v of anti-human KAPPA light chain antibody, 5~50g/L of magnesium sulfate, fatty alcohol polyoxy
1~10g/L of 0.5~10g/L of ethylene ether sodium sulfate and phenol.The pH of reagent R2 is 6~9.5.KAPPA light chain calibration object includes:
10~600mmol/L of glycine buffer, 5~50g/L of chitosan, 1~10g/L of P-hydroxybenzoic acid, Licorice root antioxidant
0.01~10g/L and KAPPA Light Chain Antigen.
The kit of the detection KAPPA light chain content of the embodiment of the present invention quickly, succinctly can be detected rapidly using simplicity
The content of KAPPA light chain in sample；It can satisfy the requirement of clinical fast high-flux detection sample, detection efficiency significantly improves
It has broad prospects in clinical application；Also have many advantages, such as that high degree of automation, detection range are wide.
The embodiment of the invention also provides a kind of kits of above-mentioned detection KAPPA light chain content in detection KAPPA
Purposes in light chain content can be used for the succinct content for rapidly detecting KAPPA light chain in sample, accuracy with higher
With good specificity.
The embodiment of the invention also provides a kind of kits using above-mentioned detection KAPPA light chain content to detect KAPPA
The method of light chain content.As shown in Figure 1, the flow chart of the method for detection KAPPA light chain content of the invention.This method includes
Following step:
Step S10: being 1~10:200 by the volume ratio of sample to be tested and reagent R1, reagent R1 be added into sample to be tested,
The first mixed liquor is obtained, and obtains the absorbance OD1 of the first mixed liquor.The step can release antigen ambient electron in sample to be tested
Layer and hydrated sheath expose antigen site sufficiently.
Step S20: it is 1:1~10 by reagent R2 and reagent R1 volume ratio, reagent R2 is added into the first mixed liquor, obtains
Second mixed liquor, and obtain the absorbance OD2 of the second mixed liquor.
Preferably, reagent R2 and reagent R1 volume ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9 or 1:
10。
Step S30: standard curve is obtained using KAPPA light chain calibration object, according to absorbance OD1 and absorbance OD2 in institute
State the content that KAPPA light chain in sample to be tested is obtained on standard curve.
Wherein, the fitting mode of standard curve include: Spline, quadratic equation, logit-log3p, logit-log4p,
Logit-log5p or Polygonal.
With specific embodiment, technical scheme is described further below.
Embodiment 1
The kit of detection KAPPA light chain content includes: reagent R1, reagent R2 and KAPPA light chain calibration object.
Wherein, reagent R1 includes following ingredient:
Reagent R2 includes following ingredient:
KAPPA light chain calibration object includes following ingredient:
The KAPPA Light Chain Antigen of corresponding amount is added to the mixed liquor of other above-mentioned reagents by calibration object concentration as required
In, it is mixed to get KAPPA light chain calibration object.The present embodiment prepares the KAPPA light chain calibration object of 5 various concentrations, and KAPPA is light
The concentration of chain antigen is respectively as follows: 0mg/dL, 200mg/dL, 400mg/dL, 800mg/dL and 1200mg/dL.By KAPPA light chain school
0.22 μm of the membrane filtration degerming of quasi- product, saves at 2~8 DEG C.
Using mentioned reagent box, sample to be tested is tested and analyzed by 7080 automatic clinical chemistry analyzer of Hitachi.Such as
It is the flow diagram of the detection method of the KAPPA light chain content of the embodiment of the present invention 1 shown in Fig. 2.Firstly, using KAPPA
Light chain calibration object passes through built-in logit-log (4p) curve matching model fitting standard curve on automatic clinical chemistry analyzer.
Then, reagent R1 is added into sample to be tested, the volume of sample to be tested is 5 μ L, and the volume of reagent R1 is 250 μ L, is mixed to get
First mixed liquor, and 5 minutes are kept the temperature at 37 DEG C, then read the absorbance OD1 of first mixed liquor.Again into the first mixed liquor
Reagent R2 is further added, reagent R2 volume is 50 μ L, obtains the second mixed liquor, and keep the temperature 5 minutes at 37 DEG C, it is mixed to read second
Close the absorbance OD2 of liquid.According to the absorbance OD2 of the absorbance OD1 of the first mixed liquor and the second mixed liquor, on standard curve
Obtain the content of the KAPPA light chain of sample to be tested.The dominant wavelength of detection is 340nm, a length of 700nm of complementary wave.
Embodiment 2
The kit of detection KAPPA light chain content includes: reagent R1, reagent R2 and KAPPA light chain calibration object.
Wherein, reagent R1 includes following ingredient:
Reagent R2 includes following ingredient:
KAPPA light chain calibration object includes following ingredient:
The KAPPA Light Chain Antigen of corresponding amount is added to the mixed liquor of other above-mentioned reagents by calibration object concentration as required
In, it is mixed to get KAPPA light chain calibration object.The present embodiment prepares the KAPPA light chain calibration object of 5 various concentrations, and KAPPA is light
The concentration of chain antigen is respectively as follows: 0mg/dL, 200mg/dL, 400mg/dL, 800mg/dL and 1200mg/dL.By KAPPA light chain school
0.22 μm of the membrane filtration degerming of quasi- product, saves at 2~8 DEG C.
Using mentioned reagent box, tested and analyzed by 7080 automatic clinical chemistry analyzer of Hitachi.Detection method is the same as real
Apply example 1.
Embodiment 3
The kit of detection KAPPA light chain content includes: reagent R1, reagent R2 and KAPPA light chain calibration object.
Wherein, reagent R1 includes following ingredient:
Reagent R2 includes following ingredient:
KAPPA light chain calibration object includes following ingredient:
The present embodiment selects high concentration single-point calibration product, therefore KAPPA Light Chain Antigen concentration is 1200mg/dL.By KAPPA
0.22 μm of the membrane filtration degerming of light chain calibration object, saves at 2~8 DEG C.When in use with normal saline dilution at 5 not
With the KAPPA light chain calibration object of concentration, the concentration of KAPPA Light Chain Antigen be respectively as follows: 0mg/dL, 200mg/dL, 400mg/dL,
800mg/dL and 1200mg/dL.
Using mentioned reagent box, tested and analyzed by 7080 automatic clinical chemistry analyzer of Hitachi.Detection method is the same as real
Apply example 1.
Embodiment 4
The kit of detection KAPPA light chain content includes: reagent R1, reagent R2 and KAPPA light chain calibration object.
Wherein, reagent R1 includes following ingredient:
Reagent R2 includes following ingredient:
KAPPA light chain calibration object includes following ingredient:
The present embodiment selects high concentration single-point calibration product, therefore KAPPA Light Chain Antigen concentration is 1200mg/dL.By KAPPA
0.22 μm of the membrane filtration degerming of light chain calibration object, saves at 2~8 DEG C.When in use with normal saline dilution at 5 not
With the KAPPA light chain calibration object of concentration, the concentration of KAPPA Light Chain Antigen be respectively as follows: 0mg/dL, 200mg/dL, 400mg/dL,
800mg/dL and 1200mg/dL.
Using mentioned reagent box, tested and analyzed by 7080 automatic clinical chemistry analyzer of Hitachi.Detection method is the same as real
Apply example 1.
Embodiment 5
Using the kit and detection method of embodiment 1, a clinical serum sample is taken at random, it is full-automatic using Hitachi 7080
Biochemical Analyzer repeats detection 10 times to same sample, and testing result is as shown in table 1.
The KAPPA light chain content (10 times) and the coefficient of variation of the detection sample of table 1
The results show that the coefficient of variation is 3.7%, less than 5%, show kit of the invention precision with higher.
Embodiment 6
Kit (using the kit of embodiment 1) of the invention and KAPPA light chain contrast agent (immunoturbidimetry, purchase
From ROCHE company) 100 clinical samples serum of blank determination.Comparison result is as shown in table 2.Kit and detection of the invention
The same embodiment of method；KAPPA light chain contrast agent sets relevant parameter to specifications, uses 7080 full-automatic biochemical of Hitachi point
Analyzer analyzes sample.
The kit of the present invention of table 2 is compared with contrast agents detected value correlation
The regression equation that linear regression obtains is Y=0.9818X+3.4795, coefficient R2=0.9952.Such as Fig. 3 institute
Show, is the comparative result schematic diagram of detection the KAPPA light chain kit and Correlation to That Abroad kit of the embodiment of the present invention 6.Fig. 3
Correlation is shown.Statistics indicate that kit of the invention and contrast agent correlation are fine, illustrate kit of the invention to inspection
Surveying KAPPA light chain content has specificity and accuracy well.
Embodiment 7
Kit (using the kit of embodiment 1) of the present invention and contrast agent box have carried out anti-interference test simultaneously,
Test result is as shown in table 3, after chaff interferent is added, to the relative errors of various chaff interferents within 10%.Contrast agents pair
The relative error of various chaff interferents is above 10%.Even if there are certain density chaff interferent including blood red egg in test sample
White, bilirubin, Vc, triglycerides etc. influence very little to the testing result of kit of the present invention.
The anti-interference test result of the reagent of the present invention of table 3
| Chaff interferent | This reagent | Relative error | Certain company's reagent | Relative error |
| Sample+water | 601 | 600 | ||
| Sample+1600mg/dl triglycerides | 624 | 3.83% | 669 | 11.50% |
| Sample+460mg/dl hemoglobin | 620 | 3.16% | 662 | 10.33% |
| Sample+50mg/dl Vc | 617 | 2.67% | 670 | 11.67% |
| Sample+18.2mg/dl bilirubin | 583 | 3.00% | 535 | 10.83% |
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc. within mind and principle should all include within protection scope of the present invention.
Claims (5)
1. a kind of kit for detecting KAPPA light chain content characterized by comprising reagent R1, reagent R2 and KAPPA light chain
Calibration object；The reagent R1 includes: glycine 400mmol/L, sodium chloride 30g/L, polyoxethylene octylphenyl phenylate 1g/L, cow's serum
The pH of albumen 1g/L, PEG4000 100g/L and Sodium azide 10g/L, the reagent R1 are 9.0；The reagent R2 includes: phosphoric acid
Salt 10mmol/L, anti-human KAPPA light chain antibody 50%w/v, sodium chloride 6g/L, sodium sulfate of polyethenoxy ether of fatty alcohol 6g/L and folded
The pH of nitrogen sodium 3g/L, the reagent R2 are 6.5；The KAPPA light chain calibration object includes: CAPSO buffer 10mmol/L, seaweed
Sugared 3g/L, Sodium azide 3g/L, dibutyl hydroxy toluene 0.5g/L and KAPPA Light Chain Antigen corresponding amount；Or,
The reagent R1 includes: phosphate 20mmol/L, sodium chloride 5g/L, polyoxyethylene alkyl phenyl ether 50g/L, cow's serum egg
The pH of white 1g/L, PEG2000 10g/L and Sodium azide 2g/L, the reagent R1 are 6.5；The reagent R2 includes: phosphate
400mmol/L, anti-human KAPPA light chain antibody 20%w/v, sodium chloride 40g/L, sodium sulfate of polyethenoxy ether of fatty alcohol 10g/L and
The pH of Sodium azide 10g/L, the reagent R2 are 8.5；The KAPPA light chain calibration object include: HEPES buffer solution 500mmol/L,
Disodium ethylene diamine tetraacetate 2g/L, mannitol 9g/L, Sodium azide 8g/L, dilauryl thiodipropionate 10g/L and KAPPA are light
Chain antigen 1 200mg/dL.
2. a kind of as described in claim 1 kit of detection KAPPA light chain content in detection KAPPA light chain content
Purposes.
3. a kind of side of the kit detection KAPPA light chain content using detection KAPPA light chain content as described in claim 1
Method characterized by comprising
It is 1~10:200 by the volume ratio of sample to be tested and the reagent R1, the reagent R1 is added in Xiang Suoshu sample to be tested,
The first mixed liquor is obtained, and obtains the absorbance OD1 of first mixed liquor；
It is 1:1~10 by the reagent R2 and the reagent R1 volume ratio, the reagent R2 is added in the first mixed liquor of Xiang Suoshu,
The second mixed liquor is obtained, and obtains the absorbance OD2 of second mixed liquor；
Standard curve is obtained using the KAPPA light chain calibration object, according to the absorbance OD1 and the absorbance OD2 in institute
State the content that KAPPA light chain in the sample to be tested is obtained on standard curve.
4. the method for detection KAPPA light chain content as claimed in claim 3, it is characterised in that: the reagent R2 and the examination
Agent R1 volume ratio is 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9 or 1:10.
5. the method for detection KAPPA light chain content as claimed in claim 3, which is characterized in that the fitting of the standard curve
Mode includes: Spline, quadratic equation, logit-log3p, logit-log4p, logit-log5p or Polygonal.
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| CN109164263A (en) * | 2018-09-05 | 2019-01-08 | 苏州普瑞斯生物科技有限公司 | A kind of kit and preparation method measuring light chain Kappa concentration |
| CN109164264A (en) * | 2018-09-05 | 2019-01-08 | 苏州普瑞斯生物科技有限公司 | A kind of kit and preparation method measuring free light chain Kappa concentration |
| CN112345474B (en) * | 2020-11-11 | 2022-06-17 | 昆明理工大学 | Method for rapidly detecting tert-butylhydroquinone in food |
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