CN102175871A - Liquid double-reagent kit for measuring free light chains in serum or urine by double-latex method - Google Patents
Liquid double-reagent kit for measuring free light chains in serum or urine by double-latex method Download PDFInfo
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Abstract
The invention relates to a liquid double-reagent kit for measuring free light chains in serum or urine by a double-latex method. The kit provided by the invention comprises a reagent R1, a reagent R2 and a light chain antigen calibrator with known concentration, wherein the reagent R1 comprises a macromolecular accelerant, a preservative, a surface active agent, a reaction promoter, a stabilizer, an electrolyte and a buffer solution; R2 comprises two types of latex particles with different diameters, wherein the diameters of the latex particles are respectively in ranges from 40nm to 80nm and from 100nm to 200nm; the two types of latex particles are both cross linked with kappa-type or lambda-type anti-FLC (full-load current) polyclonal antibody, and the latex particles comprise the preservative, the surface active agent, the reaction promoter, the electrolyte, the stabilizer and the buffer solution; the light chain antigen calibrator with known concentration comprises the preservative, the electrolyte, the stabilizer, kappa-type or lambda-type FLC antigen and the buffer solution. The kit in the invention has the advantages that the specificity is strong, the sensitivity is high, the linear range is wide, the operation is simple, and the kit is suitable for various full automatic biochemical analyzers.
Description
Technical field
The present invention relates to the invention belongs to the Medical Immunology field, relate to a kind of immunologic function test reagent, further, the present invention relates to a kind of reagent of measuring immunoglobulin (Ig) free light chain in serum or the urine.
Background technology
Immunoglobulin (Ig) (Immunoglobulin, be called for short Ig) is to have globulin general designation like antibody activity or chemical constitution and the antibody class, is present in a large number in the normal human serum, and the trace existence is also arranged in the urine.The Ig molecular structure is by " Y " type globulin of two heavy chains (Heavy chain, H chain) and two light chains (Light chain, L chain) and disulfide bond formation.People's immunoglobulin (Ig) is divided into IgM, IgG, IgA, IgD, IgE according to its heavy chain difference, and corresponding heavy chain is followed successively by μ, γ, α, δ, ε; Light chain divides κ (kappa) and λ (lambda) 2 types, and the light chain that any immunoglobulin (Ig) all contains two same types is κ type or λ type, and human total κ (kappa) and the ratio of λ (lambda) are about 2: 1.
Light chain immunoglobulin (κ or λ) is the protein by a kind of small-molecular weight (2.2kD or 4.4kD) of thick liquid cell generation.Have in the healthy human blood nearly 40% free light chain (Free light chain, FLC), the content of κ FLC and λ FLC is respectively 3～19mg/l and 6～26mg/l, κ FLC/ λ FLC is that the normal ratio of κ/λ is between 0.26～1.65.λ FLC forms dimer than κ FLC is easier by disulfide bond in addition, and the latter keeps free state usually.
The secretion of free light chain and heavy chain is a balance under the normal condition, and when thick liquid cell generation monoclonal malignant proliferation, can produce in a large number not the FLC with the heavy chain combination, has changed κ/λ ratio.If ratio＞1.65 item show that the κ light chain is excessive, the patient produces monoclonal κ light chain; If ratio＜0.26 item is that lambda light chain is excessive, the patient produces the monoclonal lambda light chain.FLC can freely pass through glomerular filtration membrane, is heavily absorbed at renal tubule and gets back in the blood circulation, so have only a small amount of FLC to exist in normal person's urine.When metabolism imbalance or generation Huppert's disease, occur lot of F LC in the blood, and, promptly produce so-called bence jones protein (Bence-Jones protein, BJP also claim M albumen) by discharging in the urine.
Bence jones protein is first special tumor marker of finding up to now, can be used to evaluation and monitoring and comprise Huppert's disease (multiple myeloma, MM), (amyloidosis, AL) etc. multiple B cell malignant proliferation disease has higher clinic value to amyloidosis.
In recent years, along with Huppert's disease (multiple myeloma, MM), amyloidosis (amyloidosis, AL) etc. multiple B cell malignant proliferation disease research deepens continuously, diagnosis index clinically is progressively by the monitoring to immunoglobulin (Ig) in the blood, the monitoring of light chain in blood or the urine (total light chain comprises the mating type of free type and immunoglobulin (Ig)), advocate up till now again at the monitoring of free light chain in the blood and calculate κ/λ ratio.Why research draws such conclusion, mainly contain following some foundation: one, to the traditional determination of immunoglobulin of monitoring rate of free light chain among the patients serum can be faster and better make diagnosis, reason is: the half life period of free light chain is very short, on average have only at 2～6 o'clock, and the half life period of immunoglobulin (Ig) is generally longer, the half life period of IgG more reaches 20-25 days, is the former hundreds of times.The free light chain level can reflect patient's the state of an illness and result of treatment in real time in theory; Two, the single mensuration at free light chain in the urine can not judge the state of an illness accurately, reason is: under the normal condition, synchronous detection arrives in urine and serum but FLC raises, if follow under the renal failure situation among the B cell malignant proliferation disease patient, light chain detects and causes mistaken diagnosis less than variation in the urine, and the mensuration of FLC can not be interfered in the serum, and is more accurate in diagnosis.And need collect patient's twenty-four-hour urine when making sample with urine, sample size is big, less stable.
Although studies show that the clinical meaning of free light chain monitoring in the blood is very high, influenced by the following aspects, the development of the mensuration of free light chain is comparatively slow in the serum.One, classic method sensitivity is lower, can not be accurately quantitative, as: electrophoresis, immunofixation electrophoresis method etc., the two emulsion methods that use among the present invention have possessed sufficiently high sensitivity; Two, specific antibody technology of preparing, the amount of immunoglobulin (Ig) is several thousand times of free light chain in the serum, the content that will accurately measure FLC in the serum will overcome the interference of a large amount of immunoglobulin (Ig)s in the serum, the selection of specific antibody is very important, if the antibody specificity difference of selecting will cause distortion as a result with the light chain generation cross-linking reaction of complete immunoglobulin (Ig).But early stage preparation at the hidden site of free light chain antibody specific recognition in conjunction with corresponding free light chain, but monoclonal antibody is tired lower, (latex enhancing method is a class of immunoturbidimetry generally not to be suitable for immunoturbidimetry, be that monoclonal antibody is unsuitable for the latex enhancing immune turbidimetry), but can make or buy antibody now by oneself, therefore make the application of latex enhancing immune turbidimetry become possibility at a plurality of hidden sites of free light chain; Three, among the normal human in the serum content of FLC very low, but its content can exceed normal tens times when the optimum or malignant proliferation pathology of B cell takes place, and therefore requires the assay method of FLC not only will possess highly sensitive, high specificity and also wants the range of linearity wide.
The latex enhancing immune turbidimetry is the wider immunoturbidimetry of a kind of application that newly-developed gets up, and has higher sensitivity than common immunoturbidimetry.Employed latex enhancing immune turbidimetry has been done again partly and has been improved among the present invention, adopt the latex particle (promptly two emulsion method) of two kinds of different-diameters, can satisfy high specificity, highly sensitive, requirement that the range of linearity is wide, can be applied to various automatic clinical chemistry analyzers, meet the requirement of clinical rapid and accurate determination.Therefore, the present invention adopts two emulsion methods, develop the liquid double reagent that can detect FLC in serum and the urine, huge using value is arranged in clinical practice, can be multiple B cell clinical diagnosises optimum or the malignant proliferation disease such as Huppert's disease, multiple sclerosis, amyloid pathology strong index is provided.According to research reports, the mensuration of free serum light chain becomes routine inspection project clinically probably in the near future, visible this index of free serum light chain critical role clinically.
Summary of the invention
The objective of the invention is to overcome the shortcoming and defect that prior art exists, and provide a kind of improvement of FLC detectable, the FLC detection kit that have that sample (serum or urine) need not to dilute, highly sensitive, high specificity, the range of linearity are wide, is applicable to various types of automatic clinical chemistry analyzers, described kit comprises the FLC calibration object of reagent R1, reagent R2 and concentration known.
On the one hand, the invention provides the kit that a kind of pair of emulsion method measured free light chain in serum or the urine, kit comprises the light chain antigen calibration object of reagent R1, reagent R2 and concentration known, wherein,
A. the reagent R1 of kit of the present invention be a kind of the special antigen site of FLC is fully exposed and can accelerate antigen-antibody reaction make it reach reaction end rapidly, comprising material accounts for its mass percent and is: 2%～8% macromolecule accelerator, 0.01%～1.0% antiseptic, 0.1%～1.0% surfactant, 0.1%～0.5% reaction promoter, 0.05～5.0% stabilizing agent, 0.1%～5.0% electrolyte, and the damping fluid of 10～100mmol/L of pH6.5～8.0;
B. the R2 of kit of the present invention comprises the latex particle of two kinds of different-diameters, its latex particle diameter range is respectively between 40～80nm and 100～200nm, all crosslinked on two kinds of latex particles have very strong κ type of specificity or an anti-FLC polyclonal antibody of λ type, can discern and in conjunction with monomer and the polymer of FLC in serum or the urine, but not combine with the mating type light chain of complete immunoglobulin (Ig).Comprise that accounting for its mass percent is: the latex particle 1.0%～5.0% of 40～80nm, the latex particle of 100～200nm, 3.0%～8.0%, 0.01%～1.0% antiseptic, 0.1%～1.0% surfactant, 0.1%～0.5% reaction promoter, 0.1%～5.0% electrolyte, 0.05～5.0% stabilizing agent, and the damping fluid of 10～100mmol/L of pH6.5～8.0;
C. the FLC calibration object of concentration known of the present invention, be used for relatively carrying out calculating as a result with sample, comprise that accounting for its mass percent is: 0.01%～1.0% antiseptic, 0.1%～5.0% electrolyte, 0.05～5.0% stabilizing agent, 0～3.0%FLC (κ type or λ type) antigen, and the damping fluid of 10～100mmol/L of pH6.5～8.0.
The anti-FLC polyclonal antibody of κ type or λ type is from mammals such as horse, sheep, rabbit, mouse among the present invention.This polyclonal antibody can be special the complete immunoglobulin (Ig) of identification on the mating type light chain hidden a plurality of antigen sites, therefore can specific bond FLC monomer and condensate, and not in conjunction with the mating type light chain on the complete immunoglobulin (Ig).Therefore, this polyclonal antibody both can ensure the efficient affinity of antigen-antibody, had also ensured the specificity of kit, had improved antijamming capability.
The macromolecule accelerator that the present invention uses can be Macrogol 2000, Macrogol 4000, Macrogol 6000, polyglycol 8000, preferred Macrogol 6000.
The reaction promoter that the present invention uses can be one or more of Macrogol 6000, polyglycol 8000, Macrogol 2000 0, hexadimethrine bromide, polybrene.
The preferred goat-anti people of the present invention FLC (κ type or λ type) antibody can be made by oneself or buy.
Latex used in the present invention is the latex particle that comprises two kinds of different-diameters, be respectively the latex particle 1.0%～5.0% of 40～80nm, the latex particle 3.0%～8.0% of 100～200nm, by allocating the proportioning of two kinds of different-diameter latex particles, reach the range of linearity of also having widened reagent when improving sensitivity.
Latex of the present invention is styrene latex, and some place abbreviates latex as with " styrene latex ", reaches common understanding in those skilled in the art, no longer is further elaborated.
Employed antiseptic is selected from Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate, ethyl mercury sodium thiosulfate among the present invention.
Employed antiseptic is a Sodium azide among the present invention.
Employed electrolyte can be kation or kation among the present invention.
The preferred sodium chloride of employed electrolyte among the present invention.
Stabilizing agent among the present invention can be made of in disodium ethylene diamine tetraacetate, magnesium chloride, bovine serum albumin(BSA), the mannitol one or more.
The preferred bovine serum albumin(BSA) of employed stabilizing agent among the present invention.
Surfactant used in the present invention is selected from one or more among polyoxyethylene laurel ether series in the non-ionic surfactant, polyoxyethylene phenyl ether, polyoxyethylene alkyl phenyl ether, Tween series, the Theist.
Employed non-ionic surfactant is Tween among the present invention.
Employed damping fluid can be phosphate buffer, TRIS buffer, HEPES damping fluid, PIPES damping fluid, glycine buffer, borate buffer solution etc. among the present invention.
Say the preferred TRIS buffer of using of damping fluid among the present invention, pH is 6.5～8.0, preferred 7.8～8.0.
The present invention adopts the latex enhancing immune turbidimetry, develops the liquid double reagent that can detect FLC in serum and the urine.Its reaction principle is: bag is by the latex particle of antibody with high specificity, combine with corresponding antigen generation specificity in the sample, form the compound of Ag-Ab-latex particle, this compound forms certain turbidity in specific damping fluid, the increase degree and the antigen concentration in the sample of turbidity are proportional, under certain wavelength, carry out turbidimetric analysis turbidimetry, can record the content of tested antigen in the sample.
In the present invention, FLC in serum or the urine (κ type or λ type) antigen combines with high specific goat-anti people FLC (κ type or the λ type) polyclonal antibody that is combined in the latex particle surface, antigen-antibody reaction takes place, the compound of the Ag-Ab-latex particle that produces in damping fluid forms certain turbidity, measure the absorbance of this turbidity at 570nm wavelength place, the reference standard curve can be obtained the content of FLC in the sample.
When the kit that adopts latex enhancing method of the present invention to measure FLC in serum or the urine is measured,, read absorbance A 1 earlier with sample and reagent R1 mixing, add R2 afterwards, read absorbance A 2, calculate the reaction absorbance, get the content of FLC in the sample at last.Further, the present invention adopts 5 scaling methods, and as computation schema, the dosage/response curve according to absorbance and calibration object are done calculates FLC content in the sample according to absorbance on dosage/response curve with splines.
Technological core of the present invention is 2 points: the one,, only at free type light chain and not in conjunction with the use of the high specific polyclonal antibody of complete immunoglobulin (Ig) mating type light chain, make in serum or urine far above special identification FLC in the presence of the immunoglobulin (Ig) of self concentration; The 2nd,, the latex particle bag quilt of antibody with high specificity and two kinds of different-diameters by regulating the proportioning of two kinds of latex, reaches the range of linearity of also having widened test when improving measurement sensitivity.
Further, the outstanding advantage that latex enhancing method of the present invention is measured FLC kit in serum or the urine is: high specificity, highly sensitive, the range of linearity is wide, easy and simple to handle, be applicable to various automatic clinical chemistry analyzers, satisfied the requirement of clinical detection greatly.
Embodiment
Embodiment one
Two emulsion methods are measured free kappa light chain kit in serum or the urine
The principal ingredient and the proportioning of kit are as follows:
Reagent R1:
TRIS buffer 20mmol/L
Sodium azide (antiseptic) 0.1%
Sodium chloride (electrolyte) 154mmol/L
PEG-6000 (macromolecule accelerator) 3%
Tween-20 (surfactant) 0.2%
Bovine serum albumin(BSA) (stabilizing agent) 0.5%
Reagent R2:
TRIS buffer 20mmol/L
Sodium azide (antiseptic) 0.1%
Sodium chloride (electrolyte) 154mmol/L
Tween-20 (surfactant) 0.2%
Bovine serum albumin(BSA) (stabilizing agent) 0.5%
Goat-anti people kappa light chain polyclonal antibody 5～8%
The latex particle 1.0%～5.0% of 40～80nm, the latex particle 3.0%～8.0% of 100～200nm.During antibody sandwich, two kinds of latex separately wrap quilt separately, guarantee that antibody sandwich is even.
Kappa light chain calibration object:
TRIS buffer 20mmol/L
Sodium azide (antiseptic) 0.1%
Sodium chloride (electrolyte) 154mmol/L
Bovine serum albumin(BSA) (stabilizing agent) 0.5%
The human serum kappa light chain 0～3% of purifying
Reagent R1 is a colourless transparent solution, and R2 is the milky white solution of homogeneous, and calibration object also is a colourless transparent solution.
When using kit of the present invention, basic parameter is set to: sample size 5.0 μ L; Reagent R1240 μ L; Reagent R280 μ L; 2 end-point methods, predominant wavelength 570nm, commplementary wave length 800nm; Calibrating mode: splines Spline; The Direction of Reaction: the reaction of rising; Measure temperature: 37 ℃.Calibrating method is: 5 calibrations.Calibrate the 1st water spot for the instrument setting, 2,3,4 is 8 times, 4 times, 2 times gained of original calibrated product dilution, and the 5th is calibration object stoste, and used dilution is a physiological saline.
Embodiment two
Two emulsion methods are measured free lambda light chain kit in serum or the urine
The principal ingredient and the proportioning of kit are as follows:
Reagent R1:
TRIS buffer 20mmol/L
Sodium azide (antiseptic) 0.1%
Sodium chloride (electrolyte) 154mmol/L
PEG-6000 (macromolecule accelerator) 4%
Tween-20 (surfactant) 0.2%
Bovine serum albumin(BSA) (stabilizing agent) 0.5%
Reagent R2:
TRIS buffer 20mmol/L
Sodium azide (antiseptic) 0.1%
Sodium chloride (electrolyte) 154mmol/L
Tween-20 (surfactant) 0.2%
Bovine serum albumin(BSA) (stabilizing agent) 0.5%
Goat-anti people Lambda light chain polyclonal antibody 5～8%
The latex particle 1.0%～5.0% of 40～80nm, the latex particle 3.0%～8.0% of 100～200nm.During antibody sandwich, two kinds of latex separately wrap quilt separately, guarantee that antibody sandwich is even.
Lambda light chain calibration object:
TRIS buffer 20mmol/L
Sodium azide (antiseptic) 0.1%
Sodium chloride (electrolyte) 154mmol/L
Bovine serum albumin(BSA) (stabilizing agent) 0.5%
The human serum Lambda light chain 0～3% of purifying
Reagent R1 is a colourless transparent solution, and R2 is the milky white solution of homogeneous, and calibration object also is a colourless transparent solution.
When using kit of the present invention, basic parameter is set to: sample size 5.0 μ L; Reagent R1240 μ L; Reagent R280 μ L; 2 end-point methods, predominant wavelength 570nm, commplementary wave length 800nm; Calibrating mode: splines Spline; The Direction of Reaction: the reaction of rising; Measure temperature: 37 ℃.Calibrating method is: 5 calibrations.Calibrate the 1st water spot for the instrument setting, 2,3,4 is 8 times, 4 times, 2 times gained of calibration object stoste dilution, and the 5th is calibration object stoste, and used dilution is a physiological saline.
Embodiment three
Correlativity, sensitivity and the range of linearity of kappa free light chain kit of the present invention
Be the better relatively superiority of reagent of the present invention, selected clinical sample is the patients serum that hospital collects, because the mensuration of urine sample is easier than serum comparatively speaking.
Contrast agents employing " free Kappa/Lambda light chain is measured kit " (see " military foundation, Wang Yusan. the problem [J] that should consider in the free serum light chain mensuration, clinical examination magazine, 2004, (22) 3:165-167. ").The principle of this contrast agents is: the latex particle bag quilt of monoclonal antibody and homogeneous, in wavelength 570nm place assaying reaction absorbance, calculate respective substance content in the sample according to calibration curve.Its R1 is the water white transparency damping fluid; R2 is the latex solution of milky monoclonal antibody sensitivity.
The experiment of 1 correlativity
Use reagent of the present invention and contrast agents to adopt Hitachi's 7180 automatic clinical chemistry analyzers that 50 parts of human serums (comprising normal and monstrosity) are measured simultaneously by each autoregressive parameter respectively, reagent parameter of the present invention is seen embodiment one.Measured value is carried out linear regression, getting regression equation is: y=1.025x+2.3421, related coefficient is: (y is a reagent of the present invention to R2=0.9935, x is a contrast agents), the result shows that reagent of the present invention and contrast agents correlativity are fine, and then explanation reagent of the present invention has excellent specificity and accuracy.This reagent is not only applicable to Hitachi's 7180 automatic clinical chemistry analyzers in addition, also can be used for the full-automatic and semi-automatic biochemical analyzer of other series such as Hitachi, Olympus, and concrete parameter can be done suitable adjustment according to major parameter.
2 sensitivity experiments
Table 1, table 2 are represented the sensitivity of reagent of the present invention and contrast agents respectively.The sensitivity of calculating 95% fiducial interval is respectively: kappa free light chain reagent 0.40mg/L of the present invention is better than contrast agents 0.84mg/L.
Table 1: reagent sensitivity of the present invention
This reagent lowest detectable limit (LLD)=12.65+3*1.90=18.35mg/L
Sensitivity: 3.6 * 18.35/164=0.40mg/L
Table 2: contrast agents sensitivity
Contrast agents lowest detectable limit (LLD)=177.35+3*21.22=241.01mg/L
Sensitivity: 3.6 * 241.01/1035.82=0.84mg/L
3 ranges of linearity
Used sample is the high value serum of a kappa free light chain patient physiological saline dilution gained, and linear error 10% is respectively with the interior range of linearity: reagent of the present invention can reach 209mg/L; Contrast agents is 180mg/L, and the result shows that the reagent range of linearity of the present invention is wideer.
Table 3: reagent of the present invention and contrast agents be the range of linearity relatively
Embodiment four
Correlativity, sensitivity and the range of linearity of lambda free light chain kit of the present invention
Be the better relatively superiority of reagent of the present invention, selected clinical sample is the patients serum that hospital collects, because the mensuration of urine sample is easier than serum comparatively speaking.
The experiment of 1 correlativity
Use reagent of the present invention and contrast agents respectively, adopt Hitachi's 7180 automatic clinical chemistry analyzers that 50 parts of human serums (comprising normal and monstrosity) are measured simultaneously by each autoregressive parameter, reagent parameter of the present invention is seen embodiment one.Measured value is carried out linear regression, and must regression equation be: y=0.9704x+4.3195, related coefficient be: R
2=0.997 (y is a reagent of the present invention, and x is a contrast agents), the result shows that reagent of the present invention and contrast agents correlativity are fine, and then explanation reagent of the present invention has excellent specificity and accuracy.This reagent is not only applicable to Hitachi's 7180 automatic clinical chemistry analyzers in addition, also can be used for the full-automatic and semi-automatic biochemical analyzer of other series such as Hitachi, Olympus, and concrete parameter can be done suitable adjustment according to major parameter.
2 sensitivity experiments
Table 4, table 5 are represented the sensitivity of reagent of the present invention and contrast agents respectively.The sensitivity of calculating 95% fiducial interval is respectively: lambda free light chain reagent 1.53mg/L of the present invention is better than contrast agents 2.97mg/L.
Table 4 lambda free light chain of the present invention reagent sensitivity
This reagent lowest detectable limit (LLD)=32.9+3*4.05=45.05mg/L
Sensitivity: 5.9*45.05/173.73=1.53mg/L
Table 5: contrast agents sensitivity
This reagent lowest detectable limit (LLD)=310.45+3*36.07=418.66mg/L
Sensitivity: 5.9*418.66/832.14=2.97mg/L
3 ranges of linearity
The method that embodiment three is same, sample is the high value serum of a lambda free light chain patient physiological saline dilution gained, calculate linear error in 10%, the linearity that draws lambda free light chain reagent of the present invention can reach 192mg/L, the linearity of contrast agents can reach 153mg/L, and reagent of the present invention has wideer linearity.
Table 6: the reagent of the present invention and the contrast agents range of linearity
Above embodiment is to explanation of the present invention and further explains, rather than limitation of the present invention, and any modification of being made in spirit of the present invention and rights protection scope all falls into protection scope of the present invention.
Claims (10)
1. two emulsion methods are measured the kit of free light chain in serum or the urine, and kit comprises the light chain antigen calibration object of reagent R1, reagent R2 and concentration known, wherein,
A. reagent R1 comprises and accounts for its mass percent and be: 2%～8% macromolecule accelerator, 0.01%～1.0% antiseptic, 0.05～5.0% stabilizing agent, 0.1%～1.0% surfactant, 0.1%～0.5% reaction promoter, 0.1%～5.0% electrolyte, and the damping fluid of 10～100mmol/L of pH6.5～8.0;
B. reagent R2 comprises the latex particle of two kinds of different-diameters, its latex particle diameter range is respectively between 40～80nm and 100～200nm, all crosslinked on two kinds of latex particles have κ type or an anti-FLC polyclonal antibody of λ type, also comprise and account for its mass percent and be: 0.01%～1.0% antiseptic, 0.1%～1.0% surfactant, 0.1%～0.5% reaction promoter, 0.1%～5.0% electrolyte, 0.05～5.0% stabilizing agent, and the damping fluid of 10～100mmol/L of pH6.5～8.0;
C. the light chain antigen calibration object of concentration known comprises and accounts for its mass percent and be: 0.01%～1.0% antiseptic, 0.1%～5.0% electrolyte, 0.05～5.0% stabilizing agent, 0～3.0% κ type or λ type FLC antigen, and the damping fluid of 10～100mmol/L of pH6.5～8.0.
2. according to claim 1 pair of emulsion method measured the kit of free light chain in serum or the urine, and wherein the anti-FLC polyclonal antibody of κ type or λ type comes from mammal, and preferred mammal comprises horse, sheep, rabbit, mouse, more preferably sheep.
3. according to claim 1 pair of emulsion method measured the kit of free light chain in serum or the urine, wherein latex comprises the latex particle of two kinds of different-diameters, is respectively and accounts for reagent R2 mass percent to be the latex particle of 40～80nm of 1.0%～5.0% and to account for the latex particle that reagent R2 mass percent is 100～200nm of 3.0%～8.0%.
4. measure the kit of free light chain in serum or the urine according to each described pair of emulsion method of claim 1-4, wherein said macromolecule accelerator is selected from Macrogol 2000, Macrogol 4000, Macrogol 6000, polyglycol 8000.
5. measure the kit of free light chain in serum or the urine according to each described pair of emulsion method of claim 1-4, wherein said antiseptic is selected from Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate or ethyl mercury sodium thiosulfate.
6. measure the kit of free light chain in serum or the urine according to each described pair of emulsion method of claim 1-4, wherein said electrolyte is kation or kation, is preferably sodium chloride.
7. measure the kit of free light chain in serum or the urine according to each described pair of emulsion method of claim 1-4, wherein said stabilizing agent is selected from one or more in disodium ethylene diamine tetraacetate, magnesium chloride, bovine serum albumin(BSA), the mannitol, is preferably bovine serum albumin(BSA).
8. measure the kit of free light chain in serum or the urine according to each described pair of emulsion method of claim 1-4, wherein said surfactant is selected from one or more among polyoxyethylene laurel ether series in the non-ionic surfactant, polyoxyethylene phenyl ether, polyoxyethylene alkyl phenyl ether, Tween series, the Theist, is preferably Tween.
9. measure the kit of free light chain in serum or the urine according to each described pair of emulsion method of claim 1-4, wherein said damping fluid is selected from phosphate buffer, TRIS buffer, HEPES damping fluid, PIPES damping fluid, glycine buffer, borate buffer solution etc., is preferably TRIS buffer.
10. measure the kit of free light chain in serum or the urine according to each described pair of emulsion method of claim 1-4, wherein said reaction promoter is selected from one or more of Macrogol 6000, polyglycol 8000, Macrogol 2000 0, hexadimethrine bromide, polybrene.
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