CN102590524B - Neutrophil gelatinase-associated lipocalin detection kit - Google Patents
Neutrophil gelatinase-associated lipocalin detection kit Download PDFInfo
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- CN102590524B CN102590524B CN201110454991.7A CN201110454991A CN102590524B CN 102590524 B CN102590524 B CN 102590524B CN 201110454991 A CN201110454991 A CN 201110454991A CN 102590524 B CN102590524 B CN 102590524B
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- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54346—Nanoparticles
Abstract
The application relates to a kind of neutrophil gelatinase-associated lipocalin detection kit, more specifically, the application relates to a kind of detection kit for neutrophil gelatinase-associated lipocalin (NGAL) concentration in human body liquid and preparation method thereof, this kit comprises reagent R1, reagent R2 and working calibration product, reagent R1 is the buffer solution of applicable antigen-antibody reaction, reagent R2 is the solution being combined with the Nano microsphere of anti-human NGAL antibody simultaneously containing size two kinds of particle diameters, this reagent is by the specific reaction of antigen-antibody, Nano microsphere is condensed, produce the change of turbidity, comparison calibration curve can quantitative measurement NGAL.This kit has higher sensitivity when measuring low NGAL concentration, having the wider range of linearity simultaneously, there will not be prozone phenomenon when measuring high NGAL concentration.This kit stable in physicochemical property, easy to use, Clinical laboratory medicine has a wide range of applications.
Description
Technical field
The application relates to a kind of neutrophil gelatinase-associated lipocalin and measures kit, more specifically, the application relates to a kind of one for neutrophil gelatinase-associated lipocalin (NGAL) concentration in human body liquid and detects reagent and preparation method thereof, this reagent comprises reagent R1, reagent R2 and working calibration product, reagent R1 is the buffer solution of applicable antigen-antibody reaction, R2 is the solution being combined with the Nano microsphere of anti-human NGAL antibody simultaneously containing size two kinds of particle diameters, this reagent is by the specific reaction of antigen-antibody, Nano microsphere is condensed, produce the change of turbidity, comparison calibration curve can quantitative measurement NGAL.
Background technology
Neutrophil gelatinase-associated lipocalin (neutrophilgelatinase-associatedlipocalin, NGAL), also referred to as Lipocalin-2 (Lipocalin-2), be a newcomer of lipocalin protein family, it is the early stage biomarker of renal dysfunction.NGAL is not only present in neutrophil cell, also appear in specific epithelial cell, such as occur in the process of ischemic and toxicity injury of kidney, NGAL in renal cells will significantly increase, in two hours that start, in urine and blood, NGAL level will significantly increase, and therefore NGAL is the Sensitive mark of early stage acute injury of kidney.
After clinical operation, can there is acute renal failure to a certain degree in the patient of more than 50%, and the morbidity later stage occurs mortality ratio is increased greatly usually.The prognosis of acute renal failure never improves, and will cause remarkable concern in 40 from now on year.The method of general diagnostic method as measured serum creatinine or cysteine proteinase inhibitor C (CystatinC) all can only detect after renal function is degenerated, and namely can only within one day, notably detect in several days after injury.And using NGAL detection kit, doctor can to make acute renal dysfunction in several hours in damage and diagnoses and do suitable process.
Research shows, in healthy volunteer's urine, NGAL content detection distribution of results is between 0.7-9.6ng/ml, and mean value is 5.3ng/ml, and the content detection result in blood plasma is 37-106ng/ml, and mean value is 63ng/ml.When after injury of kidney, NGAL level will increase suddenly rapidly.Random detection severe case, the concentration of urine NGAL be 110ng/ml to 40000ng/ml not etc., and their EDTA anticoagulate plasma testing result is 25ng/ml to 3491ng/ml.When the NGAL content in urine is more than 350ng/ml, or the positive predictive value that renal failure occurs when the NGAL content in blood plasma reaches more than 400ng/ml is 90%.
The assay method of current NGAL mainly contains euzymelinked immunosorbent assay (ELISA), radioimmunology, Western-blotting and chemoluminescence method.Euzymelinked immunosorbent assay (ELISA) automaticity is not high, and is affected by human factors larger; Radioimmunology also exists problem of environmental pollution; Western-blotting complicated operation, measures precision low; And though chemoluminescence method sensitivity is high, the mensuration range of linearity is less and testing cost is higher, and need specified chemical luminometer, these reasons cause its range of application less.
Latex-enhanced immunoturbidimetric assay is also the common method measuring NGAL concentration in human plasma or urine, and the method is less demanding to instrument and equipment, does not have the problem such as environmental protection and operating personnel's self-protection.Compare with other assay methods, the method is easy to be quick, reliably sensitive, common automatically or semi-automatic biochemical analyzer just can, have larger range of application, larger practical value.
NGAL concentration is lower than 10ng/ml in normal person's urine, and in severe nephrotic urine, concentration can up to more than 7000ng/ml.The concentration range of broadness like this, all proposes higher requirement to detection kit sensitivity and linear measurement range.Existing Latex-enhanced immunoturbidimetric assay measures the kit of NGAL concentration in human plasma or urine in the market, but these kits cannot meet the wide range of linearity and highly sensitive requirement simultaneously, therefore need diluted sample to measure high concentration NGAL, this brings inconvenience in use.
Therefore, be necessary to develop a kind of turbid detection kit of NGAL latex enhancing immune transmittance simultaneously with high sensitivity and the wide range of linearity.
Summary of the invention
Therefore, technical purpose of the present invention is that developing a kind of NGAL latex enhancing immune simultaneously with high sensitivity and the wide range of linearity projects than turbid detection kit.
Therefore, a first aspect of the present invention relates to a kind of NGAL detection kit, comprises reagent R1, reagent R2 and working calibration product, and wherein reagent R1 is NGAL and anti-human NGAL antibody response damping fluid; Reagent R2 is the Nano microsphere solution being combined with anti-human NGAL antibody; Working calibration product are made up of damping fluid, stabilizing agent, antiseptic and NGAL normative reference product, it is characterized in that, the Nano microsphere being combined with anti-human NGAL antibody of size two kinds of particle diameters is comprised in described reagent R2, Large stone is 150-500nm, small particle diameter is 50-100nm, preferably, Large stone is 200-300nm, and small particle diameter is 70-90nm.
Preferably, reagent R1 damping fluid is selected from one or more in phosphate buffer, glycine buffer, HEPES damping fluid, borate buffer, acetate buffer or ammonium chloride buffer, and its concentration is 10-500mM; Reagent R2 damping fluid used is one or more in phosphate buffer, glycine buffer, HEPES damping fluid, borate buffer, acetate buffer or ammonium chloride buffer, and its concentration is 10-500mM.
Preferably, described anti-human NGAL antibody is polyclonal antibody or monoclonal antibody, and more preferably, described polyclonal antibody is the one of rabbit anti-human NGAL antibody, goat-anti human neutrophil genatinase associated lipocalin antibody or the anti-human NGAL antibody of chicken; Monoclonal antibody is mouse-anti human neutrophil genatinase associated lipocalin antibody.
Preferably, the Nano microsphere in reagent R2 is by one or more latex be polymerized of polystyrene, acrylic acid, acrylic acid related esters class, and more preferably, comprise the latex particle of size two kinds of particle diameters in reagent R2, Large stone diameter is 150-500nm; Latex small particle diameter diameter is 50-100nm.
Preferably, in reagent R2, Nano microsphere mass percent is 0.1-1%, and wherein Large stone mass percent is 0.01-0.1%; Small particle diameter mass percent is 0.1-0.8%.
Preferably, in reagent R2, the combination of anti-human NGAL antibody and Nano microsphere is chemical crosslinking or physisorption.
Preferably, described NGAL normative reference product comprise the NGAL of five variable concentrations, are respectively: 150ng/ml, 600ng/ml, 1500ng/ml, 3000ng/ml, 5000ng/ml, or five of similar ratio variable concentrations; Wherein damping fluid is HEPES pH of buffer 7.4, and stabilizing agent is one or more mixing of BSA, sucrose, sodium chloride, polysorbas20.
Most preferably, described NGAL protein detection kit is selected from following combination:
Kit 1: reagent R1 consists of: pH7.3, Main Ingredients and Appearance is:
Glycocoll 50mM,
NaCl0.15M，
NaN30.1％，
EDTA10mM，
BSA0.1％，
Rabbit igg 1%;
R2 consists of:
PH7.0, principal ingredient is:
HEPES20mM，
NaN30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%,
Working calibration product are: buffer matrix, and it is by HEPES50mM, NaCl0.15M, NaN30.1%, EDTA10mM, BSA0.5%, sucrose 5%, and all the other are deionized water composition, are dissolved in the NGAL normative reference product of the 5000ng/ml concentration of described matrix;
Kit 2: reagent R1 consists of: pH7.4, Main Ingredients and Appearance is:
Glycocoll 50mM,
NaCl0.25M，
NaN30.1％，
EDTA10mM，
BSA0.1％，
R2 consists of: pH7.0
HEPES20mM，
NaN30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%,
Working calibration product are: buffer matrix, and it is by HEPES50mM, NaCl0.15M, NaN30.1%, EDTA10mM, BSA0.5%, polysorbas20 0.1%, and all the other are deionized water composition, are dissolved in the NGAL normative reference product of the 5000ng/ml concentration of described matrix;
Kit 3: reagent R1 consists of: pH7.4, Main Ingredients and Appearance is:
Glycocoll 50mM,
NaCl0.15M，
NaN30.1％，
EDTA10mM，
BSA0.1％，
Mouse IgG1%,
R2 consists of: pH7.0
HEPES20mM，
NaN30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%,
Working calibration product are: buffer matrix, and it is by HEPES50mM, NaCl0.15M, NaN30.1%, EDTA10mM, BSA0.5%, sucrose 5%, and all the other are deionized water composition, are dissolved in the NGAL normative reference product of the 5000ng/ml concentration of described matrix;
A second aspect of the present invention relates to the preparation method of NGAL detection kit as above, comprise step: be blended in pH7.0 damping fluid by Large stone Nano microsphere and anti-human NGAL antibody-solutions, stirring at room temperature 3 hours, antibody is combined with Nano microsphere fully, centrifugally remove supernatant, after adding damping fluid, ultrasonic disperse is even; Be blended in pH7.0 damping fluid by small particle diameter Nano microsphere and anti-human NGAL antibody-solutions, stirring at room temperature 3 hours, antibody is combined with Nano microsphere and centrifugally fully removes supernatant, and after adding damping fluid, ultrasonic disperse is even; Size particles solution is mixed according to a certain percentage.
In other words, the application's problem to be solved is the defect overcoming the existence of above-mentioned available reagent box, there is provided a kind of latex enhancing immune of size particles compound than turbid kit, detect the content of NGAL in blood plasma or urine, to reach the object of high sensitivity and the wide range of linearity simultaneously.
Possess the object of high sensitivity and the wide range of linearity for reaching a kind of kit, the application mainly starts with from immune response speed simultaneously, namely under low NGAL concentration, have higher immune agglutination speed, ensures high sensitivity; Simultaneously under high NGAL concentration, lower immune agglutination speed be had, ensure the wide range of linearity.
Specifically, the inventive concept of the application is: after (1) large grain size latex pan coating antibody, and itself and antigen compatibility are higher, antigen-antibody reaction speed when drastically increasing low concentration NGAL, thus improves the sensitivity of reagent; (2) after small particle diameter pan coating antibody, its compatibility is relatively low, under high antigen concentration, still can form antigen antibody complex, thus improves the range of linearity of reagent; (3), after large small particle diameter is blended in same buffer solution system, reach high sensitivity and wide range of linearity object simultaneously.
The technical scheme of the application is:
A kind of NGAL detection kit, it is characterized in that, comprise reagent R1, reagent R2, described reagent R1 is the damping fluid of applicable NGAL and anti-human NGAL antibody response; Described reagent R2 is the Nano microsphere being combined with anti-human NGAL antibody, is placed in suitable damping fluid.
Described R1 damping fluid is one or more in phosphate buffer, glycine buffer, HEPES damping fluid, borate buffer, acetate buffer or ammonium chloride buffer.Its concentration is 10-500mM; Reagent R2 damping fluid is one or more in phosphate buffer, glycine buffer, HEPES damping fluid, borate buffer, acetate buffer or ammonium chloride buffer.Its concentration is 10-500mM.
Described anti-human NGAL antibody is polyclonal antibody or monoclonal antibody.
Described polyclonal antibody is rabbit anti-human NGAL antibody, goat-anti human neutrophil genatinase associated lipocalin antibody or the anti-human NGAL antibody of chicken.
Described monoclonal antibody is mouse-anti human neutrophil genatinase associated lipocalin antibody.
Nano microsphere in described reagent R2 is by one or more latex be polymerized of polystyrene latex, acrylic acid, acrylic acid related esters class.
Latex in described reagent R2 comprises the latex of size two kinds of particle diameters.Bulky grain diameter is 150-500nm; Latex small particle diameters is 50-100nm.
In described reagent R2, latex quality percentage is 0.1-1%, and wherein bulky grain mass percent is 0.01-0.1%; Latex granule mass percent is 0.1-0.8%.
In described reagent R2, anti-human NGAL antibody and latex particle combination are physisorption or chemical crosslinking.
The major advantage of the application is: (1) bulky grain latex surface hydrophobicity is higher, and after coated antibody, itself and antigen compatibility are higher, antigen-antibody reaction speed when drastically increasing low concentration NGAL, thus improves the sensitivity of reagent.(2) after granule pan coating antibody, its compatibility is relatively low, under high antigen concentration, still can form antigen antibody complex, thus improves the range of linearity of reagent; (3) after size particles is blended in same buffer solution system; due to bulky grain and NGAL and antigen adhesion strong; therefore be first combined with antigen; like this under low NGAL concentration; reagent also has higher sensitivity; when NGAL concentration is higher, antigen can be combined with granule and form immune complex, and such reagent has the wide range of linearity.Utilize the Nano microsphere of size two kinds of particle diameters to realize the report improving detectability and expand the range of linearity detected although prior art also has simultaneously, but prior art size particles adopts conventional bag by mode, higher for antibody performance requirement, some needs two kinds of antibody, a kind of high-affinity antibody of some needs, and the coated antibody mode that the application is different because size particles adopts, antagonist does not specially require, and only needs a kind of antibody just.
Accompanying drawing explanation
Fig. 1: NGAL kit standard curve.
Fig. 2: the present invention and enzyme-linked immunosorbent assay blood plasma measured value correlativity.
Fig. 3: the present invention and enzyme-linked immunosorbent assay urine measured value correlativity.
Embodiment
To further illustrate the present invention by following non-limiting example below, as well known to those skilled in the art, without departing from the spirit of the invention, can make many amendments to the present invention, such amendment also falls into scope of the present invention.Such as, as well known to those skilled in the art, can use the NGAL antibody that different plant species is originated, the particle size range of the Nano microsphere of described two kinds of particle diameters also can be wider, and such change also all falls within protection scope of the present invention.
Following experimental technique if no special instructions, is conventional method, the experiment material used as antibody and Nano microsphere if no special instructions, all can easily obtain from commercial company.
Embodiment
The preparation of embodiment 1:NGAL detection kit
The application has exemplarily prepared three kinds of NGAL detection kit, and its preparation raw material, preparation steps, performance are as follows:
(1) kit 1:
R1：pH7.3，
Main Ingredients and Appearance is:
Glycocoll 50mM,
NaCl0.15M，
NaN30.1％，
EDTA10mM，
BSA0.1％，
Rabbit igg 1%,
R2 preparation process is as follows:
Prepared by small grain size latex reagent:
1, at room temperature diluting the anti-human NGAL polyclonal antibody (10mg/ml) of 4ml rabbit with 20mMHEPES solution (pH7.0), is 2mg/ml to antibody final concentration;
2, at room temperature dilute 4ml74nm latex solution (10%wt) with 20mMHEPES solution (pH7.0), make latex concentration be 4%;
3, the antibody diluted and latex are mixed, room temperature adds the EDAC solution 10ml preparing 5mg/ml with 20mMHEPES solution (pH7.0) after shaking 30 minutes, room temperature reaction 3h, then adds 5ml confining liquid (1%BSA, 1% polysorbas20 solution), close 2h;
4, centrifugal, abandon supernatant, by 20mMHEPES solution (pH7.0) diluted latex to 2.5%, and add 0.1% antiseptic, stand-by after ultrasonic disperse.
Prepared by large grain size latex reagent:
1, at room temperature diluting the anti-human NGAL polyclonal antibody (10mg/ml) of 4ml rabbit with 20mMHEPES solution (pH7.0), is 2mg/ml to antibody final concentration;
2, at room temperature dilute 1ml220nm latex solution (10%wt) with 20mMHEPES solution (pH7.0), make latex concentration be 4%;
3, the antibody diluted and latex are mixed, room temperature adds the EDAC solution 10ml preparing 5mg/ml with 20mMHEPES solution (pH7.0) after shaking 30 minutes, room temperature reaction 3h, then adds 5ml confining liquid (containing 1%BSA, 1% polysorbas20 solution), close 2h;
4, centrifugal, abandon supernatant, by 20mMHEPES solution (pH7.0) diluted latex to 2.5%, and add 0.1% antiseptic, stand-by after ultrasonic disperse latex particle.
5, large and small particle reagents is mixed with final R2 reagent: after size particles being mixed, stir, R2 prepares complete:
R2：pH7.0，
Principal ingredient is:
HEPES20mM，
NaN30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%.
It is as follows that normative reference savors damping fluid matrix components:
HEPES50mM，
NaCl0.15M，
NaN30.1％，
EDTA10mM，
BSA0.5％，
Sucrose 5%,
All the other are deionized water,
By concentration required for normative reference product, restructuring NGAL sterling is added in above-mentioned damping fluid matrix, the NGAL normative reference product of obtained 5000ng/ml concentration, the normative reference product (150ng/ml, 600ng/ml, 1500ng/ml, 3000ng/ml) of all the other concentration can be obtained by 5000ng/ml standard items damping fluid diluted matrix.
(2) kit 2:
R1：pH7.4，
Main Ingredients and Appearance is:
Glycocoll 50mM
NaCl0.25M
NaN30.1％
EDTA10mM
BSA0.1％
R2 preparation process is as follows:
Prepared by small grain size latex reagent:
1, at room temperature diluting the anti-human NGAL polyclonal antibody (3mg/ml) of 5ml chicken with 20mMHEPES solution (pH7.0), is 2mg/ml to antibody final concentration;
2, at room temperature dilute 4ml74nm latex solution (10%wt) with 20mMHEPES solution (pH7.0), make latex concentration be 4%;
3, the antibody diluted and latex are mixed, room temperature adds the EDAC solution 10ml preparing 5mg/ml with 20mMHEPES solution (pH7.0) after shaking 30 minutes, room temperature reaction 3h, then adds 5ml confining liquid (1%BSA, 1% polysorbas20 solution), close 2h;
4, centrifugal, abandon supernatant, by 20mMHEPES solution (pH7.0) diluted latex to 2.5%, and add 0.1% antiseptic, stand-by after ultrasonic disperse.
Prepared by large grain size latex reagent:
1, at room temperature diluting the anti-human NGAL polyclonal antibody (5mg/ml) of 4ml chicken with 20mMHEPES solution (pH7.0), is 2mg/ml to antibody final concentration;
2, at room temperature dilute 1ml220nm latex solution (10%wt) with 20mMHEPES solution (pH7.0), make latex concentration be 4%;
3, the antibody diluted and latex are mixed, room temperature adds the EDAC solution 10ml preparing 5mg/ml with 20mMHEPES solution (pH7.0) after shaking 30 minutes, room temperature reaction 3h, then adds 5ml confining liquid (containing 1%BSA, 1% polysorbas20 solution), close 2h;
4, centrifugal, abandon supernatant, by 20mMHEPES solution (pH7.0) diluted latex to 2.5%, and add 0.1% antiseptic, stand-by after ultrasonic disperse latex particle.
5, large and small particle reagents is mixed with final R2 reagent: after size particles being mixed, stir, R2 prepares complete.
R2：pH7.0，
Principal ingredient is:
HEPES20mM，
NaN30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%.
It is as follows that normative reference savors damping fluid matrix components:
HEPES50mM，
NaCl0.15M，
NaN30.1％，
EDTA10mM，
BSA0.5％，
Polysorbas20 0.1%,
All the other are deionized water,
By concentration required for normative reference product, restructuring NGAL sterling is added in above-mentioned damping fluid matrix, the NGAL normative reference product of obtained 5000ng/ml concentration, the normative reference product (150ng/ml, 600ng/ml, 1500ng/ml, 3000ng/ml) of all the other concentration can be obtained by 5000ng/ml standard items damping fluid diluted matrix.
(3) kit 3:
R1：pH7.4，
Main Ingredients and Appearance is:
Glycocoll 50mM,
NaCl0.15M，
NaN30.1％，
EDTA10mM，
BSA0.1％，
Mouse IgG1%,
R2 preparation process is as follows:
Prepared by small grain size latex reagent:
1, at room temperature diluting 5ml mouse-anti human neutrophil genatinase associated lipocalin polyclonal antibody (5mg/ml) with 20mMHEPES solution (pH7.0), is 2mg/ml to antibody final concentration;
2, at room temperature dilute 4ml74nm latex solution (10%wt) with 20mMHEPES solution (pH7.0), make latex concentration be 4%;
3, the antibody diluted and latex are mixed, room temperature adds the EDAC solution 10ml preparing 5mg/ml with 20mMHEPES solution (pH7.0) after shaking 30 minutes, room temperature reaction 3h, then adds 5ml confining liquid (1%BSA, 1% polysorbas20 solution), close 2h;
4, centrifugal, abandon supernatant, by 20mMHEPES solution (pH7.0) diluted latex to 2.5%, and add 0.1% antiseptic, stand-by after ultrasonic disperse.
Prepared by large grain size latex reagent:
1, at room temperature diluting 4ml mouse-anti human neutrophil genatinase associated lipocalin polyclonal antibody (5mg/ml) with 20mMHEPES solution (pH7.0), is 2mg/ml to antibody final concentration;
2, at room temperature dilute 1ml220nm latex solution (10%wt) with 20mMHEPES solution (pH7.0), make latex concentration be 4%;
3, the antibody diluted and latex are mixed, room temperature adds the EDAC solution 10ml preparing 5mg/ml with 20mMHEPES solution (pH7.0) after shaking 30 minutes, room temperature reaction 3h, then adds 5ml confining liquid (containing 1%BSA, 1% polysorbas20 solution), close 2h;
4, centrifugal, abandon supernatant, by 20mMHEPES solution (pH7.0) diluted latex to 2.5%, and add 0.1% antiseptic, stand-by after ultrasonic disperse latex particle.
5, large and small particle diameter reagent is mixed with final R2 reagent: after size particles being mixed, stir, R2 prepares complete.
R2：pH7.0，
Principal ingredient is:
HEPES20mM，
NaN30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%.
It is as follows that normative reference savors damping fluid matrix components:
HEPES50mM
NaCl0.15M
NaN30.1％
EDTA10mM
BSA0.5％
Polysorbas20 0.1%
Sucrose 5%
All the other are deionized water
By concentration required for normative reference product, restructuring NGAL sterling is added in above-mentioned damping fluid matrix, the NGAL normative reference product of obtained 5000ng/ml concentration, the normative reference product (0ng/ml, 150ng/ml, 600ng/ml, 1500ng/ml, 3000ng/ml) of all the other concentration can be obtained by 5000ng/ml standard items damping fluid diluted matrix.
Embodiment 2:NGAL detection kit determination step, the drafting of calibration curve, carries out comparative study with the product of certain famous brand name on market, this producer's reagent Cleaning Principle is also latex enhancing immune turbidimetry, mainly forms: R1,10mmolTris, 0.1%NAN3,0.5%BSA; R2, the 0.15% latex suspension being coated with anti-human NGAL monoclonal antibody, contrast situation is as follows:
The determination step of the application is as shown in table 1.
Table 1: the determination step of the application's kit:
For the kit 2 of embodiment 1-(2), standard concentration is transverse axis, corresponding Δ OD
570for the longitudinal axis, adopt nonlinear fitting, as spline draws out typical curve, result as shown in Figure 1.
Linear and the lowest detectable limit of embodiment 3:NGAL kit
Linear Experiment: adopt method known to those skilled in the art, a certain high NGAL concentration samples (urine specimen during acute kidney injury after surgery patient 2h) is done doubling dilution, with the kit 2 shown in embodiment 1-(2) for detection kit, measure the concentration after dilution, measure and average for three times, compared with theoretical concentration, calculate its deviation from linearity.Contrast with certain famous brand name on above-mentioned market, measurement result is as shown in table 2, table 3, table 4 and table 5 simultaneously.
The high level scope of table 2 the application kit 2 is linear: unit ng/ml
Table 3: the high level scope of contrast agents is linear: unit ng/ml
| Extension rate | Measure 1 | Measure 1 | Measure 1 | Measure average | Theoretical value | Deviation from linearity % |
| 1 | 4102.3 | 4056.5 | 4085.6 | 4081.467 | 6839.523 | 40.33 |
| 0.9 | 4110.5 | 4125.4 | 4108.7 | 4114.867 | 6162.513 | 33.23 |
| 0.8 | 4178.4 | 4165.2 | 4182.6 | 4175.4 | 5485.503 | 23.88 |
| 0.7 | 4034.5 | 3955.1 | 3927.5 | 3972.367 | 4808.493 | 17.39 |
| 0.6 | 3893.6 | 3851.8 | 3906.6 | 3884 | 4131.483 | 5.99 |
| 0.5 | 3426 | 3466.5 | 3439.3 | 3443.933 | 3454.473 | 0.305 |
| 0.4 | 2793.6 | 2781.5 | 2815.5 | 2796.867 | 2777.463 | -0.7 |
| 0.3 | 2071.2 | 2039.1 | 2063.4 | 2057.9 | 2100.453 | 2.026 |
| 0.2 | 1486.1 | 1499.5 | 1491.9 | 1492.5 | 1423.443 | -4.85 |
| 0.1 | 712.9 | 714.6 | 705.7 | 711.0667 | 746.433 | 4.738 |
Table 4: the low value scope of the application's kit 2 is linear: unit ng/ml
| Extension rate | Measure 1 | Measure 1 | Measure 1 | Measure average | Theoretical value | Deviation from linearity % |
| 1 | 720.3 | 720.5 | 720.6 | 720.47 | 711.07 | 1.321 |
| 0.9 | 630.5 | 632.4 | 638.7 | 633.87 | 639.12 | -0.82 |
| 0.8 | 568.4 | 565.2 | 562.6 | 565.4 | 567.18 | -0.31 |
| 0.7 | 504.5 | 495.1 | 492.5 | 497.37 | 495.23 | 0.43 |
| 0.6 | 419.6 | 415.8 | 420.6 | 418.67 | 423.29 | -1.09 |
| 0.5 | 346 | 346.5 | 349.3 | 347.27 | 351.35 | -1.16 |
| 0.4 | 278.6 | 271.5 | 280.5 | 276.87 | 279.40 | -0.91 |
| 0.3 | 206.2 | 209.1 | 203.4 | 206.23 | 207.46 | -0.59 |
| 0.2 | 141.1 | 140.5 | 141.9 | 141.17 | 135.51 | 4.17 |
| 0.1 | 65.7 | 68.6 | 69.7 | 68 | 63.57 | 6.97 |
| 0.05 | 25.5 | 25.6 | 25.4 | 25.5 | 27.60 | -7.60 |
| 0 | 0.02 | -0.01 | 0.01 | 0.01 | 0 | - |
Table 5: the low value scope of contrast agents is linear: unit ng/ml
| Extension rate | Measure 1 | Measure 1 | Measure 1 | Measure average | Theoretical value | Deviation from linearity % |
| 1 | 740.3 | 740.5 | 740.6 | 740.4667 | 731.066 | -1.29 |
| 0.9 | 650.5 | 652.4 | 658.7 | 653.8667 | 659.122 | 0.797 |
| 0.8 | 588.4 | 585.2 | 582.6 | 585.4 | 587.178 | 0.303 |
| 0.7 | 524.5 | 515.1 | 512.5 | 517.3667 | 515.234 | -0.41 |
| 0.6 | 439.6 | 435.8 | 440.6 | 438.6667 | 443.29 | 1.043 |
| 0.5 | 366 | 366.5 | 369.3 | 367.2667 | 371.346 | 1.099 |
| 0.4 | 298.6 | 291.5 | 300.5 | 296.8667 | 299.402 | 0.847 |
| 0.3 | 226.2 | 229.1 | 223.4 | 226.2333 | 227.458 | 0.538 |
| 0.2 | 161.1 | 160.5 | 161.9 | 161.1667 | 155.514 | -3.63 |
| 0.1 | 85.7 | 88.6 | 89.7 | 88 | 83.57 | -5.3 |
| 0.05 | 45.5 | 45.6 | 45.4 | 45.5 | 47.598 | 4.408 |
| 0 | 740.3 | 740.5 | 740.6 | 740.4667 | 731.066 | -1.29 |
The range of linearity of the application's kit 2 is 0-7000ng/ml as can be seen from the above table, meanwhile, the test result of two other kit of the present invention and the result of kit 2 similar.The range of linearity of contrast agents box is only 0-3800ng/ml.
Lowest detectable limit: adopt method known to those skilled in the art, blank solution and same portion physiological saline are carried out the several low concentration samples diluted, with the kit 2 shown in embodiment 1-(2) for detection kit measures, replication 15 times, reads absorbance variable quantity.Then the absorbance of each sample after deduction blank absorbency is calculated, computation of mean values, standard deviation.The possibility of 99.7% is adopted to calculate lowest detectable limit.The average of each sample is deducted the respective standard deviation of 3 times, then compare with the standard deviation of 3 times of blank solution, if the former is higher than the latter, just assert that the minimum absorbance having the possibility of 99.7% to occur necessarily is greater than blank absorbency, can quantitative reporting the result.Measurement result is as shown in table 6-9.
Table 6: the lowest detectable limit of the application's kit 2:
Table 7: the absorbance that said determination result is corresponding
| (A-A0)*10000 | 1 | 2 | 3 | Water is blank | |
| 1 | 41 | 24 | 15 | 8 | 1 |
| 2 | 38 | 22 | 15 | 5 | 6 |
| 3 | 35 | 22 | 12 | 9 | 4 |
| 4 | 29 | 23 | 10 | 5 | 0 |
| 5 | 34 | 19 | 13 | 6 | 2 |
| 6 | 42 | 19 | 14 | 3 | 3 |
| 7 | 36 | 22 | 14 | 4 | 1 |
| 8 | 39 | 22 | 15 | 1 | 1 |
| 9 | 40 | 23 | 14 | 3 | 3 |
| 10 | 34 | 24 | 16 | 7 | -1 |
| 11 | 42 | 22 | 14 | 4 | 2 |
| 12 | 37 | 21 | 17 | 5 | 6 |
| 13 | 39 | 22 | 11 | 8 | 0 |
| 14 | 44 | 21 | 12 | 4 | -1 |
| 15 | 41 | 23 | 16 | 4 | 5 |
| Average | 38.06667 | 21.93333 | 13.86667 | 5.066667 | 2.133333 |
| SD | 3.954503 | 1.486447 | 1.959106 | 2.186539 | 2.325838 |
| CV％ | 10.38836 | 6.777113 | 14.12817 | 43.15537 | 109.0237 |
| 3SD | 11.86351 | 4.45934 | 5.877317 | 6.559617 | 6.977515 |
| A-3SD | 26.20316 | 17.47399 | 7.989349 | -1.49295 | 9.110848 |
| A-3SD&3S0 | 17.09231 | 8.363145 | -1.1215 | -10.6038 |
Table 8: the lowest detectable limit of contrast agents
Table 9: the absorbance that said determination result is corresponding
Learnt by above table, the lowest detection of the application's kit is limited to 10.39ng/ml, simultaneously the test result of two other kit of the present invention and the result of kit 2 similar.Contrast agents is 14.67ng/ml.
Embodiment 4: the correlativity of the application NGAL kit and enzyme-linked immunosorbent assay (Elisa) measured value
Carry out two kinds of sample correlations to investigate: EDTA anticoagulate plasma, urine specimen.
The EDTA anticoagulate plasma NGAL measured value obtained with embodiment 1-(2) with compared by the measured value of enzyme-linked immunosorbent assay gained, as shown in Figure 2, to go forward side by side line retrace analysis, know correlativity R=0.9982 (y=0.9646x+4.7736); Demonstrate the present embodiment and enzyme-linked immunosorbent assay has good correlativity in EDTA anticoagulate plasma NGAL mensuration.
The urine NGAL measured value obtained with the described kit of embodiment 1-(2) with compared by the measured value of enzyme-linked immunosorbent assay gained, as shown in Figure 3, to go forward side by side line retrace analysis, know correlativity R=0.9998 (y=0.9794x-9.0372), this demonstrates this kit 2 of the present invention and in urine NGAL mensuration, also has good correlativity with enzyme-linked immunosorbent assay.Meanwhile, the test result of two other kit of the present invention and the result of kit 2 similar.
Claims (6)
1. a NGAL detection kit, comprises:
Reagent R1, reagent R2 and working calibration product;
Wherein reagent R1 is NGAL and anti-human NGAL antibody response damping fluid; Reagent R2 is the Nano microsphere solution being combined with anti-human NGAL antibody; Working calibration product are made up of damping fluid, stabilizing agent, antiseptic and NGAL normative reference product,
Comprise the Nano microsphere being combined with anti-human NGAL antibody of size two kinds of particle diameters in wherein said reagent R2, Large stone Nano microsphere is 220nm, and small particle diameter Nano microsphere is 74nm;
Wherein said NGAL detection kit is selected from as follows:
Kit 1:
Reagent R1 consists of:
pH7.3，
Glycocoll 50mM,
NaCl0.15M，
NaN
30.1％，
EDTA10mM，
BSA0.1％，
Rabbit igg 1%;
R2 consists of:
pH7.0，
HEPES20mM，
NaN
30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%,
Working calibration product are:
Buffer matrix, it is by HEPES50mM, NaCl0.15M, NaN
30.1%, EDTA10mM, BSA0.5%, sucrose 5%, all the other are deionized water composition, are dissolved in the NGAL normative reference product of the 5000ng/ml concentration of described matrix;
Kit 2:
Reagent R1 consists of:
pH7.4，
Glycocoll 50mM,
NaCl0.25M，
NaN
30.1％，
EDTA10mM，
BSA0.1％，
R2 consists of:
pH7.0
HEPES20mM，
NaN
30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%,
Working calibration product are:
Buffer matrix, it is by HEPES50mM, NaCl0.15M, NaN
30.1%, EDTA10mM, BSA0.5%, polysorbas20 0.1%, all the other are deionized water composition, are dissolved in the NGAL normative reference product of the 5000ng/ml concentration of described matrix;
Kit 3:
Reagent R1 consists of:
pH7.4，
Glycocoll 50mM,
NaCl0.15M，
NaN
30.1％，
EDTA10mM，
BSA0.1％，
Mouse IgG1%,
R2 consists of:
pH7.0
HEPES20mM，
NaN
30.1％，
Small grain size latex content is: 2%,
Large grain size latex content is: 0.5%,
Working calibration product are:
Buffer matrix, it is by HEPES50mM, NaCl0.15M, NaN
30.1%, EDTA10mM, BSA0.5%, sucrose 5%, all the other are deionized water composition, are dissolved in the NGAL normative reference product of the 5000ng/ml concentration of described matrix.
2. NGAL protein detection kit according to claim 1, is characterized in that, described anti-human NGAL antibody is polyclonal antibody or monoclonal antibody.
3. NGAL detection kit according to claim 2, is characterized in that,
Described polyclonal antibody is the one of rabbit anti-human NGAL antibody, goat-anti human neutrophil genatinase associated lipocalin antibody or the anti-human NGAL antibody of chicken;
Monoclonal antibody is mouse-anti human neutrophil genatinase associated lipocalin antibody.
4. the NGAL detection kit according to any one of claims 1 to 3, is characterized in that, the Nano microsphere in reagent R2 is by one or more latex be polymerized of polystyrene, acrylic acid, acrylic acid related esters class.
5. the NGAL detection kit according to any one of Claims 1-4, is characterized in that, in reagent R2, the combination of anti-human NGAL antibody and Nano microsphere is chemical crosslinking or physisorption.
6. the preparation method of the NGAL detection kit according to any one of claim 1 to 5, comprises step:
Be blended in pH7.0 damping fluid by Large stone Nano microsphere and anti-human NGAL antibody-solutions, stirring at room temperature 3 hours, antibody is combined with Nano microsphere fully, centrifugally removes supernatant, and after adding damping fluid, ultrasonic disperse is even;
Be blended in pH7.0 damping fluid by small particle diameter Nano microsphere and anti-human NGAL antibody-solutions, stirring at room temperature 3 hours, antibody is combined with Nano microsphere and centrifugally fully removes supernatant, and after adding damping fluid, ultrasonic disperse is even;
Size particles solution is mixed according to a certain percentage;
Wherein Large stone Nano microsphere is 220nm, and small particle diameter Nano microsphere is 74nm, and Large stone Nano microsphere content is 0.5%, and small particle diameter Nano microsphere content is 2%.
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| CN201110454991.7A CN102590524B (en) | 2011-12-30 | 2011-12-30 | Neutrophil gelatinase-associated lipocalin detection kit |
| PCT/CN2012/086508 WO2013097606A1 (en) | 2011-12-30 | 2012-12-13 | Assay kit for neutrophil gelatinase-associated lipocalin |
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| CN201110454991.7A CN102590524B (en) | 2011-12-30 | 2011-12-30 | Neutrophil gelatinase-associated lipocalin detection kit |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0898169A2 (en) * | 1997-08-11 | 1999-02-24 | F. Hoffmann-La Roche Ag | Microparticle enhanced light scattering assay and microparticle reagents therefor |
| CN1769894A (en) * | 2004-11-01 | 2006-05-10 | 上海申索佑福医学诊断用品有限公司 | Kit for determination of high-sensitive C-reactive protein |
| CN101566633A (en) * | 2009-04-17 | 2009-10-28 | 武汉华美生物工程有限公司 | Method for diagnosing, evaluating or testing cancer and foreseeing cancer severity |
| CN101680891A (en) * | 2007-11-21 | 2010-03-24 | 爱科来株式会社 | Measurement reagent, immune nephelometry using the same, and analyte analysis tool |
| CN102175871A (en) * | 2010-12-30 | 2011-09-07 | 北京九强生物技术股份有限公司 | Liquid double-reagent kit for measuring free light chains in serum or urine by double-latex method |
| CN102253217A (en) * | 2011-04-07 | 2011-11-23 | 武汉生之源生物科技有限公司 | Detection kit of latex particle enhanced neutrophil gelatinase-associated lipid transfer protein |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9182391B2 (en) * | 2008-05-09 | 2015-11-10 | Arkray, Inc. | Method of producing insoluble carrier particles, insoluble carrier particles, measurement reagent, specimen analyzing tool, and immunoturbidimetric assay |
| US7977110B2 (en) * | 2008-06-02 | 2011-07-12 | Children's Hospital Medical Center | Method for distinguishing between kidney dysfunctions |
| CN102072960A (en) * | 2010-03-29 | 2011-05-25 | 武汉生之源生物科技有限公司 | Method for detecting neutrophil gelatinase-associated lipocalin (NGAL) in sample |
| CN102243241A (en) * | 2011-04-07 | 2011-11-16 | 武汉生之源生物科技有限公司 | Homogeneous phase aerosol particle-type neutrophile granulocyte gelatinase-related lipid carrier protein determination kit and preparation method thereof |
| CN102680698B (en) * | 2011-07-29 | 2016-03-30 | 南京诺尔曼生物技术有限公司 | Neutrophil gelatinase-associated lipocalin (NGAL) measures kit (latex enhancing immune turbidimetry) |
| CN102590524B (en) * | 2011-12-30 | 2016-01-06 | 北京九强生物技术股份有限公司 | Neutrophil gelatinase-associated lipocalin detection kit |
| CN102662059B (en) * | 2012-05-11 | 2014-08-13 | 北京美康生物技术研究中心 | Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof |
-
2011
- 2011-12-30 CN CN201110454991.7A patent/CN102590524B/en active Active
-
2012
- 2012-12-13 WO PCT/CN2012/086508 patent/WO2013097606A1/en active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0898169A2 (en) * | 1997-08-11 | 1999-02-24 | F. Hoffmann-La Roche Ag | Microparticle enhanced light scattering assay and microparticle reagents therefor |
| CN1769894A (en) * | 2004-11-01 | 2006-05-10 | 上海申索佑福医学诊断用品有限公司 | Kit for determination of high-sensitive C-reactive protein |
| CN101680891A (en) * | 2007-11-21 | 2010-03-24 | 爱科来株式会社 | Measurement reagent, immune nephelometry using the same, and analyte analysis tool |
| CN101566633A (en) * | 2009-04-17 | 2009-10-28 | 武汉华美生物工程有限公司 | Method for diagnosing, evaluating or testing cancer and foreseeing cancer severity |
| CN102175871A (en) * | 2010-12-30 | 2011-09-07 | 北京九强生物技术股份有限公司 | Liquid double-reagent kit for measuring free light chains in serum or urine by double-latex method |
| CN102253217A (en) * | 2011-04-07 | 2011-11-23 | 武汉生之源生物科技有限公司 | Detection kit of latex particle enhanced neutrophil gelatinase-associated lipid transfer protein |
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