CN108761070A - A kind of urine transferrins detection kit of width detection range - Google Patents
A kind of urine transferrins detection kit of width detection range Download PDFInfo
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Abstract
The invention discloses a kind of urine transferrins detection kit of wide detection range, the kit includes reagent one and reagent two, and the reagent one includes metal ion additive, and the metal ion additive is zinc salt, a concentration of 6-19mM of the zinc salt；The reagent two is the latex particle solution for being marked with UTRF antibody, and the latex particle particle size range is 172-200nm.The kit range of linearity upper limit in the present invention is more than 80mg/L, can cover 90% or more sufferer, and reproducible near 2mg/L reference values, is conducive to gray area sample and judges；In addition, the kit in the present invention can substitute import reagent, sufferer spending is saved.
Description
Technical field
The present invention relates to biotechnologies, more particularly to a kind of urine transferrins detection reagent of wide detection range
Box.
Background technology
Transferrins (UTRF) is negatively charged small molecule circulating protein, transferrins and albumin under physiological status
All it is difficult to by glomerulus filter membrane, but when lesion occurs for kidney, transferrins and albumin can be leaked out from glomerulus, therefore
Urinary metabolites are the sensitive indications of a reflection glomerulus filter membrane damage.The uniqueness of transferrins not only makes it have detection
Conventional renal function characteristic, and more early stage, the detection of slighter injury of kidney can also be applied more broadly in.
Normal adult's urine transfer Ferritin Levels are relatively low, usually<2mg/L, but transferrins in normal adult's blood
Content is up to 2200-4000mg/L, and when slight damage occurs in glomerulus, transferrins will drain in urine in blood so that
UTRF in urine is significantly raised；Wherein, middle and later periods nephrosis sufferer, UTRF leak out more so that the UTRF concentration in urine is high
Up to hundreds of even 1000mg/L.The immunoturbidimetry reagent of common detection blood transferrin clinical at present, because sensitivity is asked
Topic can not detect urine transferrins.Now You Duo companies possess the reagent for measuring urine UTRF on the market, remove Siemens
Outside the import urine transferrins detection reagent of Beckman Kurt；Meikang biology, hundred Ao Taikang of Beijing biologies and this public affairs
Department is all made of the Monitoring lower-cut of latex immunoturbidimetry, wherein Meikang biology, hundred Ao Taikang of Beijing biologies and our company's reagent
Can 0.6mg/L be reached hereinafter, still in these reagents, the range of linearity is highest to only reach 30mg/L, and most producers
Antigen excess range is not marked also.
In clinical application, the problem that existing latex immunoturbidimetry reagent has the range of linearity narrow, the study found that
50% or more early stage glomerulonephritis sufferer, the UTRF in urine can be more than 30mg/L, only less than 10% early stage kidney
It can be more than 80mg/L that bead nephrosis, which suffers from the UTRF in urine,.Therefore, existing to utilize latex immunoturbidimetry reagent, detect urine
The kit of middle UTRF cannot be satisfied the detection of UTRF contents in most of early stage glomerulonephritis sufferer urines at all, and a large amount of
Sample using the dilution reinspection of artificial or instrument, not only increased the labor intensity of hospital staff but increase reagent loss at
This.
Currently, the mainstream reagent of UTRF contents is Siemens BNII (2.2-35mg/L) system of import in detection urine
UTRF reagents in UTRF reagents and Beckman Kurt (BC) IM800 (2.0-40mg/L) system, both using scattering
Immunoturbidimetry can reach 35-40mg/L although its detection range is relatively wide, cover 60% sufferer, it is examined
Lower bound is surveyed to be only capable of reaching 2.2 and 2.0mg/L.Since the sample less than minimum detection limit can not be reported as a result, and this purpose is joined
It is 2mg/L to examine value then, causes clinical application relatively awkward, gray area sample can not detect as a result, weakly positive sample is (in urine
UTRF contents are 2.0-5mg/L) measure poor repeatability.In addition, import reagent is expensive, and in existing detection urine
The kit of UTRF, detection speed is extremely slow, respectively less than 250t/h, can not adapt to that large hospital sufferer is more and physical examination magnanimity
The demand of sample.
Detection range is expanded, will be reduced by instrument pre-dilution mode than turbid reagent based on existing UTRF latex
The biochemical instruments detection speed of 50%-70%, relatively existing Siemens and BC reagents, the promotion of the method detection speed are simultaneously unknown
It is aobvious, and the ability of which expansion detection range is limited, is only capable of reaching 50-60mg/L, it is super can not to reach the range of linearity upper limit
Cross the requirement of 80mg/L.In addition, antibody dosage is increased substantially, it is simple although can be used for widening the reagent range of linearity
Antibody dosage is improved, antigen excess range can not only be adversely affected, but also can significantly promote testing cost,
Country keep under strict control medical expense background under, the method that increases substantially antibody dosage to widen the reagent range of linearity, it is clear that be nothing
What method was promoted the use of.
Therefore it provides a kind of range of linearity upper limit is more than 80mg/L, 90% or more sufferer can be covered, and join in 2mg/L
It is neighbouring reproducible to examine value, is conducive to the domestic urine transferrins detection kit that gray area sample judges, is people in the art
The problem of member's urgent need to resolve.
Invention content
In view of this, the present invention provides a kind of urine transferrins detection kit of wide detection range, linear model
Limit is placed more than 80mg/L, 90% or more sufferer can be covered, and reproducible near 2mg/L reference values, is conducive to gray area
Sample judges；In addition, the kit in the present invention can substitute import reagent, sufferer spending is saved.
To achieve the goals above, the present invention adopts the following technical scheme that：
A kind of urine transferrins detection kit of width detection range, which is characterized in that the kit includes reagent
One and reagent two, the reagent one includes metal ion additive.
Urine transferrins detection kit in the present invention, in the case where antigen excess range meets 1000mg/L,
The range of linearity upper limit is more than 80mg/L, and gray area sample meets the detection of 2mg/L concentration, and its precision CV<2%, it disclosure satisfy that
90% or more sufferer is detected；In addition, the kit in the present invention can substitute import reagent completely, disease is significantly saved
Suffer from spending.
Further, the metal ion in the metal ion additive includes iron ion, copper ion or zinc ion.
Iron ion, copper ion or zinc ion can significantly improve as the metal ion in metal ion additive and turn iron
The low side sensitivity of albumen test.The degree of saturation of transferrins coupled ion may have an impact its protein conformation, that is, influence
It forms the ability of antigen antibody complex in immunoturbidimetry measurement, and saturation degree is higher, and it is stronger to form turbidity ability.In reagent
A certain amount of metal ion is added in one, low value sample transferrin content is low, and the saturation degree of coupled ion is high, and turbidity is brought to believe
Number value is high, conversely, high concentration sample transferrin content is high, relative ion combination saturation degree is low, so the change of turbidity Forming ability
Change unobvious.
Further, the metal ion additive can be combined and soluble easily in water with transferrins；The metal ion adds
It includes zinc salt to add agent, a concentration of 6-19mM of the zinc salt, a concentration of 10-15mM of the preferred zinc salt.
Further, the zinc salt is zinc sulfate, the one or several kinds in zinc chloride or zinc acetate
Wherein, the metal ion additive further includes molysite and mantoquita, and the molysite is ferrous sulfate；The mantoquita is
Copper sulphate.
Although molysite, mantoquita or zinc salt can be used as the metal ion additive in reagent one, ferrous sulfate and sulphur
Sour copper is larger to the Color influences of one solution of reagent, and relative cost is higher；And to the face of one solution of reagent after zinc salt dissolving
Color influences less, and zinc salt Application in Chemical Engineering is more, and cost is relatively low, therefore it is preferred that using zinc salt as metal ion additive.
Further, the reagent two is the latex particle solution for being marked with UTRF antibody；The latex particle grain size model
It encloses for 172-200nm；The preferred latex particle particle size range is 185-200nm.
Using the smaller latex of grain size, reagent sensitivity can be integrally reduced, utilizes sensitivity and linear and antigen excess model
Contradictory theoretical foundation is enclosed, the range of linearity can be expanded, but directly uses the smaller latex of grain size, low side sensitivity will be caused
It is low, it leads to the problem of and gray area sample cannot be distinguished.Even if adding sensitizer, whole raising reagent sensitivity is carried out, but promoted low
It holds sensitivity simultaneously, high level sensitivity signal value can be caused excessively high, reduce the range of linearity and antigen excess range.
Therefore, in the detection of urine UTRF, when using low value loss of sensitivity caused by the smaller latex of grain size, with
The proper metal ion that reagent one is added, the promotion of the low value sensitivity brought, when cancelling out each other, and the promotion is smaller
High level sensitivity is influenced, could can not only expand the range of linearity of urine UTRF kits, but also low value sensitivity can be maintained, to
Ensure the accuracy of gray area sample measures.
Further, the reagent one further includes the sodium chloride or potassium chloride of a concentration of 400mM/L；Concentration is not higher than 20g/
The dispersant of the sensitizer of L and a concentration of 5ml/L.Wherein, although effect phase of the potassium chloride with sodium chloride in this kit
Together, but potassium chloride is with high costs compared to sodium chloride, therefore the preferred sodium chloride of the present invention.
Preferably, in reagent one further include the reagents such as preservative and buffer solution；Wherein preservative include Sodium azide or
Proclin series；The pH value of buffer solution is 5.5-8.0, including citric acid, and Good ' s, phosphate, glycine or Tris are buffered
Liquid.The present invention can appropriately add preservative and buffer solution according to actually detected situation, wherein UTRF to the variation of pH value simultaneously
It is insensitive, therefore the addition of the reagents such as preservative and buffer solution has no effect on the detection performance of kit of the present invention.
Further, the sensitizer be polyethylene glycol PEG4000, polyethylene glycol PEG6000, polyethylene glycol PEG8000,
One or more of polyethylene glycol PEG12000 or polyethylene glycol PEG20000.
It is a kind of " immune ratio of raising latex that the present invention also utilizes Patent No. 201710986970.7, patent name simultaneously
Quadratic convergence polymeric latex renovation technique, feeds intake not improving antibody disclosed in the method for turbid method antigen excess and the range of linearity "
Volume, that is, cost is constant, does not reduce detection speed without using instrument pre-dilution, antigen excess range meets the feelings of 1000mg/L
Under condition, the raising of the urine UTRF detection kit ranges of linearity in the present invention is realized.Wherein Patent No. 201710986970.7
The patent of invention of (a method of improve latex immunoturbidimetry antigen excess and the range of linearity), although quadratic convergence is utilized
Polymeric latex renovation technique feeds intake volume not improving antibody, does not use instrument pre-dilution, and antigen excess range meets
In the case of 1000mg/L, the range of linearity of UTRF detection reagents can be extended to 0.2-40mg/L, reach import reagent
The range of linearity, and low value sensitivity is better than import reagent, but clinical demand can not be fully met.Kit phase in the present invention
Compared with Patent No. 201710986970.7 (a method of improve latex immunoturbidimetry antigen excess and the range of linearity)
Patent of invention, in the case where antigen excess range meets 1000mg/L, it can be achieved that the range of linearity upper limit of kit is more than
80mg/L, gray area sample meet the detection of 2mg/L concentration, and its precision CV<2%, disclosure satisfy that 90% or more sufferer into
Row detection.
It can be seen via above technical scheme that compared with prior art, the present invention provides a kind of urine of wide detection range
Liquid transferrins detection kit has following technological merit：
(1) metal ion additive is added in the kit in the present invention in reagent one, and has selected suitable latex
Grain particle size range, low value sensitivity caused by using the smaller latex of grain size loss and keep the low value that metal ion is brought sensitive
The promotion of degree is cancelled out each other, and the promotion less influences on high level sensitivity, can could both expand the line of kit of the present invention
Property range, and low value sensitivity can be maintained, to ensure the accuracy of gray area sample measures.
(2) kit in the present invention, in the case where antigen excess range meets 1000mg/L, the range of linearity upper limit is super
80mg/L is crossed, gray area sample meets the detection of 2mg/L concentration, and its precision CV<2%, it disclosure satisfy that 90% or more sufferer
It is detected；
(3) kit using the present invention is detected, alternative import reagent, saves sufferer spending.
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technology description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 attached drawings are influence of the various concentration zinc acetate provided by the invention to measurement various concentration UTRF sensitivity.
Fig. 2 attached drawings are influence of the various concentration ferrous sulfate provided by the invention to measurement various concentration UTRF sensitivity.
Fig. 3 attached drawings are influence of the various concentration copper sulphate provided by the invention to measurement various concentration UTRF sensitivity.
Specific implementation mode
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation describes, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Embodiment 1
It is right using the method described in Patent No. 201710986970.7, i.e. quadratic convergence polymeric latex renovation technique
Our company's first generation UTRF reagents expand its range of linearity, obtain reagent A, and verify the property of first generation UTRF reagents and reagent A
Energy.
First generation UTRF reagents：Registration certificate Shanghai tool note standard 20162400121, range of linearity 0.2-24mg/L, antigen
Surplus ranging from 1000mg/L；Main constituents are：
Reagent one：Sodium chloride, a concentration of 400mM/L；Polyethylene glycol PEG6000, a concentration of 1g/L；Tween-20, it is a concentration of
5ml/L；
Reagent two：Latex particle, 3g/L, latex particle size 297nm；UTRF antibody (article No. Q0327), 10ml/L, brand
Dako。
Wherein, the progress quadratic convergence polymeric latex transformation of first generation UTRF reagents is as follows：
(1) use is identical with first generation UTRF reagents, and the Polymicrospheres companies article No. of 500ml is
The latex solution of CB0297C, wherein including 10% mass concentration, grain size is the latex of 297nm, passes through Millipore companies
50KD retention aperture ultrafiltration membrane packet be added distilled water carry out cross-flow filtration, complete 8 times change liquid.
(2) it by the latex solution after displacement distilled water, is added in four mouthfuls of reaction kettles that capacity is 1000ml, 1.5g is added
Lithium chloride and 0.4g neopelexes, reaction kettle are placed in thermostat water bath, keep 50 DEG C of constant temperature, 260rpm machineries
Stir 10min.After then passing to argon gas removal oxygen 5min, 0.15g potassium peroxydisulfates, the secondary convergence caused inside microballoon is added
Polymerisation continues in 50 DEG C of constant temperature, reacts 10h under 260rpm mechanical agitations.
(3) by complete to restrain the latex solution after polymerisation be added 50mM/L concentration, PH5.0 MES-NaOH solution,
Aperture ultrafiltration membrane packet is retained by the 50KD of Millipore companies and carries out cross-flow filtration, is completed 16 times and is changed liquid.
Restrain the grain size D of latex in latex solution before polymerisation1For 297nm, it is transformed by quadratic convergence polymeric latex
Afterwards, the latex particle size in latex solution is D2=0.78D1=231nm.
The main constituents of reagent A are：
Reagent one：Sodium chloride, a concentration of 400mM/L；Polyethylene glycol PEG6000, a concentration of 1g/L；Tween-20, it is a concentration of
5ml/L；Preservative PC-300, a concentration of 0.35ml/L；
Reagent two：Latex particle after quadratic convergence polymerization, a concentration of 3g/L, latex particle size D2(D2=0.78D1)
231nm；UTRF antibody (article No. Q0327), 10ml/L, brand Dako use conventional chemical coupling method reagent preparation two.
Simultaneously first generation UTRF reagents and reagent A, use 2ul samples, 105ul reagents one, 35ul reagents two,
570nm wavelength verifies performance, specific data such as table 1-3 in the case of 18-34 read points on Hitachi7180.
1 first generation UTRF reagent calibration data of table
| mg/L | 0 | 1.5 | 3 | 6 | 12 | 24 |
| ΔABS | 6 | 433 | 912 | 1990 | 4161 | 9374 |
2 A reagent calibration data of table
| mg/L | 0 | 2 | 10 | 20 | 40 |
| ΔABS | 2 | 275 | 1533 | 3471 | 7592 |
The comparison of table 3 first generation UTRF reagents and the reagent A range of linearity and antigen excess range
By table 1-3 it is found that reagent A is with respect to first generation UTRF reagents, the range of linearity upper limit of reagent A, which can be extended to, is more than
40mg/L, antigen excess range reach 1000mg/L.
Embodiment 2
Verify the effect of metallic addition
Using reagent A, the zinc acetate of various concentration, copper sulphate and ferrous sulfate are added respectively in the reagent one of reagent A,
Use 2ul samples, 105ul reagents one, 35ul reagents two, in Hitachi7180 in the case of 570nm wavelength, 18-34 read points
On, Δ ABS is measured using the UTRF standard items of same batch various concentration, as a result as shown in table 4-6 and Fig. 1-3.
Influence of the zinc acetate of 4 various concentration of table to measurement various concentration UTRF sensitivity
Influence of the ferrous sulfate of 5 various concentration of table to measurement various concentration UTRF sensitivity
Influence of the copper sulphate of 6 various concentration of table to measurement various concentration UTRF sensitivity
From table 4-6 and Fig. 1-3 it is found that adding zinc acetate, copper sulphate or ferrous sulfate in the reagent one of reagent A, on a small quantity
Metal ion can significantly improve the sensitivity (about promoting 120%) that low concentration UTRF is measured, as concentration of metal ions increases,
The sensitivity enhancement of low concentration (2mg/L) UTRF has tended towards stability (variation of 6 third example 2mg/L concentration sensitivities of table), middle height
Concentration (such as 10mg/L) UTRF measurement sensitivities increase with metal ion additive amount and start to gradually increase, but metallic addition
Excessive concentration when, such as when 24mM/L, ABS break bounds when the range of linearity is 40mg/L, cannot meet the range of linearity upper limit is more than
The requirement of 40mg/L.In addition, in all cases, metallic addition, the range of linearity are added in the reagent of reagent A one merely
It is unable to reach 80mg/L, the equal break bounds of ABS.
Wherein, it is light blue that the reagent one of copper sulphate, which is added, and the reagent one that ferrous sulfate is added is light green color, is not met
The most of reagents one in this field are the requirement of colourless or faint yellow clear solution, and cost is slightly higher, not as preferred, subsequent implementation
Example is colourless using solution and industrializes low-cost zinc salt.
Embodiment 3
Using this field routine thinking, reduce latex particle size in a certain range, expands the reagent range of linearity.
By the latex that Polymicrospheres companies grain size in 1 reagent A of embodiment is 297nm, it is changed to Bangslab
Grain size is the latex of 200nm, the quadratic convergence polymeric latex renovation technique described in Patent No. 201710986970.7,
Obtain D3The latex of=0.78 × 200nm=156nm, obtains reagent B, and the main composition of reagent B is divided into：
Reagent one：Sodium chloride, a concentration of 400mM/L；Polyethylene glycol PEG6000, a concentration of 1g/L；Tween-20, it is a concentration of
5ml/L；Preservative PC-300, a concentration of 0.35ml/L；
Reagent two：Latex particle after quadratic convergence polymerization, a concentration of 3g/L, latex particle size D3For 156nm；UTRF antibody
(article No. Q0327), a concentration of 10ml/L, brand Dako use conventional chemical coupling method reagent preparation two.
In the reagent one of reagent B, different strength acetic acid zinc is added, the reagent two of reagents B, using 2ul samples,
105ul reagents one, 35ul reagents two use same batch in the case of 570nm wavelength, 18-34 read points on Hitachi7180
The UTRF standard items of secondary various concentration measure Δ ABS, the neighbouring precision of reference value, the range of linearity and antigen excess range, as a result
As shown in table 7-9.
Influence of the zinc acetate of 7 various concentration of table to Δ ABS in various concentration UTRF
Influence of the zinc acetate of 8 various concentration of table to measurement reference value concentration precision
| Zinc acetate concentration | mean(mg/L) | SD | CV |
| 0mM | 2.03 | 0.173 | 8.52% |
| 6mM | 2.02 | 0.032 | 1.58% |
| 12mM | 2.03 | 0.033 | 1.63% |
| 19mM | 2.01 | 0.03 | 1.49% |
| 24mM | 2.06 | 0.031 | 1.50% |
Influence of the zinc acetate of 9 various concentration of table to antigen excess range and the range of linearity
The present embodiment reduction latex particle size is to 200nm, the secondary receipts described in using Patent No. 201710986970.7
Polymeric latex renovation technique is held back, and the zinc acetate of 6-19mM is added in the reagent of reagent B one.From table 8 it is found that the present embodiment
In reagent can realize, in the precision CV of 2mg/L<2%；From table 7 and table 9 it is found that reagent in the present embodiment, linearly
Range limit can be more than 80mg/L, and antigen excess range meets the requirement of 1000mg/L, for the UTRF reagents of the wide range of linearity.Together
When, as zinc acetate concentration improves, sensitivity is also gradually promoted when reagent measures high level, but antigen excess range is under gradually
Drop；When zinc acetate concentration is 6-12mM, it is even more that antigen excess range is up to 1200mg/L；But when zinc acetate concentration is
When 19mM, theoretical antigen excess ranging from 1000mg/L, but the range of linearity of concentration mensuration only has 80.16mg/L at this time,
Antigen excess volume edges in the 80mg/L ranges of linearity；If continue to improve zinc acetate concentration to 24mM, it is in 80mg/
The ABS break bounds of the L ranges of linearity can no longer meet the requirement that the range of linearity reaches 80mg/L.
Embodiment 4
Verify the lower limit of latex particle size
Latex in 3 reagent B of embodiment is changed to latex (the Polymicrospheres companies goods that grain size is 172nm
Number it is CB0172E), because overall sensitivity declines more, low value sensitivity requirement is cannot be satisfied by zinc acetate merely, therefore adjust increasing
Quick dose is polyethylene glycol PEG8000, concentration 20g/L, and the quadratic convergence described in Patent No. 201710986970.7 is poly-
Latex renovation technique is closed, D is obtained4The latex of=0.78 × 172nm=134nm, further obtains reagent C, main group of reagent C
Ingredient is：
Reagent one：Sodium chloride, a concentration of 400mM/L；Polyethylene glycol PEG8000, a concentration of 20g/L；Tween-20, concentration
For 5ml/L；Preservative PC-300, a concentration of 0.35ml/L；
Reagent two：Latex particle after quadratic convergence polymerization, a concentration of 3g/L, latex particle size D4For 134nm；UTRF antibody
(article No. Q0327), a concentration of 10ml/L, brand Dako use conventional chemical coupling method reagent preparation two.
In the reagent one of reagent C, the zinc acetate of various concentration is added, the reagent two in reagents C uses 2ul samples
This, 105ul reagents one, 35ul reagents two, in the case of 570nm wavelength, 18-34 read points on Hitachi7180, use is identical
The UTRF standard items of batch various concentration measure Δ ABS, the neighbouring precision of reference value, the range of linearity and antigen excess range, knot
Fruit is as shown in table 10-12.
Influence of the zinc acetate of 10 various concentration of table to Δ ABS in various concentration UTRF
Influence of the zinc acetate of 11 various concentration of table to measurement reference value concentration precision
| Zinc acetate concentration | mean(mg/L) | SD | CV |
| 0mM | 1.92 | 0.221 | 11.51% |
| 6mM | 2.01 | 0.039 | 1.94% |
| 12mM | 1.99 | 0.038 | 1.91% |
| 19mM | 2.02 | 0.04 | 1.98% |
| 24mM | 1.99 | 0.039 | 1.96% |
Influence of the zinc acetate of 12 various concentration of table to antigen excess range and the range of linearity
By table 10-12 it is found that the present embodiment is reducing latex particle size to 172nm, Patent No. is used
In 201710986970.7, in the case of the quadratic convergence polymerization of record, and in reagent 1 be added 6-19mM zinc acetate
When, 2mg/L precision CV may be implemented<2%, it can be more than 80mg/L that the range of linearity, which is reached the standard grade, and antigen excess range meets 1000mg/
The wide range of linearity UTRF reagents that L is required.It can be seen that when latex particle size is reduced to 172nm, polyethylene glycol PEG8000
As conventional sensitizer, it is already added to higher concentration i.e. 20g/L, this concentration is close to be easy to bring PEG protein precipitations non-specific
The concentration 25g/L of reaction, then latex particle size is reduced, precision CV has been difficult to ensure.In the second of 6-19mM in the present embodiment
Under sour zinc concentration, the latex particle size scope of application is 172-200nm.
In addition it can see from the comparison of embodiment 3 and 4, latex particle size improves, and the range of linearity and antigen excess range will be big
Amplitude declines.The latex particle size of 200nm has been located under the conditions of 19mM zinc acetate concentrations and 1g/L PEG6000 in embodiment 3
In antigen excess volume edges.
The present disclosure provides a kind of urine transferrins detection kits of wide detection range, by being tried in kit
Being added in agent one can be combined with transferrins, and metal ion additive soluble easily in water, and select suitable latex particle
Particle size range, make the promotion of the low value sensitivity that metal ion brings with using low value sensitivity caused by the smaller latex of grain size
Loss cancel out each other, and the promotion of low value sensitivity less influences on high level sensitivity, meets in antigen excess range
In the case of 1000mg/L, the range of linearity for having expanded kit in the present invention is reached the standard grade more than 80mg/L, and can maintain low value
Sensitivity, to ensure that gray area sample meets the detection of 2mg/L concentration, and its precision CV<2%, it disclosure satisfy that 90% or more
Sufferer be detected, using the present invention kit substitute import reagent, can significantly save sufferer spending.
Each embodiment is described by the way of progressive in this specification, the highlights of each of the examples are with other
The difference of embodiment, just to refer each other for identical similar portion between each embodiment.For device disclosed in embodiment
For, since it is corresponded to the methods disclosed in the examples, so description is fairly simple, related place is said referring to method part
It is bright.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest range caused.
Claims (7)
1. a kind of urine transferrins detection kit of width detection range, which is characterized in that the kit includes reagent one
With reagent two, the reagent one includes metal ion additive.
2. a kind of urine transferrins detection kit of wide detection range according to claim 1, which is characterized in that institute
It includes iron ion, copper ion or zinc ion to state the metal ion in metal ion additive.
3. a kind of urine transferrins detection kit of wide detection range according to claim 1, which is characterized in that institute
Stating metal ion additive can be combined and soluble easily in water with transferrins；The metal ion additive includes zinc salt, the zinc
A concentration of 6-19mM of salt.
4. a kind of urine transferrins detection kit of wide detection range according to claim 3, which is characterized in that institute
It is zinc sulfate, the one or several kinds in zinc chloride or zinc acetate to state zinc salt.
5. a kind of urine transferrins detection kit of wide detection range according to claim 1, which is characterized in that institute
It is the latex particle solution for being marked with transferrin antibodies to state reagent two；The particle size range of the latex particle is 172-200nm.
6. a kind of urine transferrins detection kit of wide detection range according to claim 1, which is characterized in that institute
State the sodium chloride or potassium chloride that reagent one further includes a concentration of 400mM/L；Concentration is not higher than the sensitizer of 20g/L and a concentration of
The dispersant of 5ml/L.
7. a kind of urine transferrins detection kit of wide detection range according to claim 1, which is characterized in that institute
It is Tween-20 to state dispersant；The sensitizer is polyethylene glycol PEG4000, polyethylene glycol PEG6000, polyethylene glycol
One or more of PEG8000, polyethylene glycol PEG12000 or polyethylene glycol PEG20000.
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| CN114047338A (en) * | 2021-11-10 | 2022-02-15 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
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| CN114047338A (en) * | 2021-11-10 | 2022-02-15 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
| CN114047338B (en) * | 2021-11-10 | 2024-03-12 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
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