CN207248894U - Bladder chalone C time resolution detection card and kit - Google Patents
Bladder chalone C time resolution detection card and kit Download PDFInfo
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- CN207248894U CN207248894U CN201721358604.9U CN201721358604U CN207248894U CN 207248894 U CN207248894 U CN 207248894U CN 201721358604 U CN201721358604 U CN 201721358604U CN 207248894 U CN207248894 U CN 207248894U
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Abstract
A kind of sensitivity using time-resolved fluoroimmunoassay chromatographic technique is the utility model is related to, realizes high sensitivity with reference to dry type immunofluorescence analysis instrument, can accurate quantitative analysis, simple and efficient bladder chalone C time-resolved fluoroimmunoassay chromatography quantitative test card and kit.The bladder chalone C time-resolved fluorescence detection card includes the interior test strip that gets stuck that gets stuck and be arranged on, the test strip is made of the end liner and glass fiber sample pad being pasted onto on end liner, nitrocellulose filter and blotting paper, and the sample pad, nitrocellulose filter and blotting paper, which sequentially overlap, to be pasted onto on end liner.The utility model detection card and kit can in accurate quantitative analysis detection human serum, blood plasma and whole blood sample bladder chalone C content, utilize time-resolved fluorescence microballoon detection technique, sample fluorescence itself can be avoided to disturb, microballoon is connected by mark by covalent bond with antibody, marked product is stablized, and has the characteristics that wide detection range, high sensitivity, accuracy is high, detection is fast and convenient.
Description
(1) technical field
It the utility model is related to a kind of bladder chalone C time-resolved fluorescence detection card and kit.
(2) background technology
Bladder chalone C (cystatin c) is a kind of cystatin, also referred to as γ-trace of albumin and
γ-postglobulin, is widely present in the karyocyte and body fluid of various tissues, is a kind of low molecular weight, alkaline both the non-glycated egg
White matter, molecular weight 13.3KD, is made of 122 amino acid residues, can be produced by all karyocytes of body, generation rate is permanent
It is fixed.Bladder chalone C in circulation is only eliminated through glomerular filtration, is a kind of endogenous mark for reflecting glomerular filtration rate(GFR change
Will thing, and in proximal convoluted tubule reabsorption, but blood is not returned to by complete metabolic breakdown after reabsorption, therefore, its blood level by
Glomerular filtration determines, and does not depend on any foeign element, is a kind of reflection glomerulus filter such as gender, the influence at age, diet
Cross the preferable homology marker of rate change.
Under normal circumstances, concentration of the CysC in serum and blood plasma is 0.51-1.09mg/L (term of reference).Work as renal function
When impaired, the concentration of CysC in blood changes and changes with glomerular filtration rate(GFR.During renal failure, glomerular filtration rate(GFR declines,
Concentration can increase more than 10 times to CysC in blood;If glomerular filtration rate(GFR is normal, and when renal tubular function is not normal, CysC can be hindered
Absorb in renal tubule and decompose rapidly, the concentration in urine is increased more than 100 times.Research data is it has been shown that CysC is compared with serum
BUN, Cr have the Sensitivity and Specificity of higher, have very important value for evaluation glomerular filtration rate(GFR, CysC is so far
Substantially meet the endogenous material of preferable endogenous GFR marker requirements.It is the one kind for the assessment renal function that newly-developed gets up
The index that sensitiveness is good, specificity is high.
The method for measuring bladder chalone C in the prior art mainly has immunoturbidimetry, fluorescence immune chromatography method, colloidal gold method
Deng.Wherein colloidal gold method has the advantages that easy to operate quick, but sensitivity is low and quantitative inaccurate;Immunoturbidimetry is more quick
It is sense, accurate, full automatic biochemical apparatus is can be applied to, but instrument and time-consuming longer is needed, it is adapted to processing great amount of samples, can not meets fast
The purpose of speed detection;Immunofluorescence technique is that bladder chalone C antibody is covalently bonded on fluorescent microsphere surface active groups, by swashing
Whether detection line produces fluorescence and carrys out judging result after hair, rapid and convenient can accurate quantitative analysis have the high sensitivity, label steady at the same time
The advantages that determining, in medical immunology detection field extensive use.
But many compounds and albumen in biofluid and serum can inherently fluoresce, therefore use tradition
Fluorescein when chromophore carries out fluoroscopic examination sensitivity will degradation, but most of background fluorescence signal is to deposit in short-term
, come labelled antibody or antigen as label using rare-earth fluorescent microballoon (lanthanide chelate).Because of lanthanide series
Chelate stoke displacements are easy to differentiate exciting light greatly and launch light, so that exciting light interference is excluded, and decay time length can lead to
Cross time resolution and eliminate background fluorescence interference, in addition lanthanide chelate exciting light spectrum width and emission spectrum is narrow further to carry
High sensitivity and reduction background fluorescence.Therefore time-resolved fluoroimmunoassay technology, while two ginsengs of Detection wavelength and time are used
Number carries out signal resolution, can effectively exclude the interference of non-specific fluorescence, drastically increase sensitivity for analysis.
(3) content of the invention
The utility model aim is to provide a kind of sensitivity using time-resolved fluoroimmunoassay chromatographic technique, with reference to dry type
Immunofluorescence analysis instrument realize high sensitivity, can accurate quantitative analysis, simple and efficient bladder chalone C time-resolved fluoroimmunoassay chromatography it is fixed
Amount detection card and kit.
The technical solution adopted in the utility model is:
A kind of bladder chalone C time-resolved fluorescence detection card, including get stuck and be arranged on the interior test strip that gets stuck:
The test strip is by the end liner and glass fiber sample pad being pasted onto on end liner, nitrocellulose filter and suction
Water paper forms, and the sample pad, nitrocellulose filter and blotting paper, which sequentially overlap, to be pasted onto on end liner;
The sample pad is provided with microballoon line, is coated with the bladder chalone C monoclonal antibody of time-resolved fluorescence microballoon mark
CysC1 (content is the μ l fluorescent microspheres of 50~200 μ g antibody/200), the rare earth doped europium element of time-resolved fluorescence microballoon
(solid content 1%, w/w), particle diameter are 100nm~300nm;The time-resolved fluorescence microballoon is stablized under ground state, 350~
Launch fluorescence of the wave-length coverage in 600~620nm under the excitation source effect of 380nm;
Detection line and nature controlling line are disposed with the nitrocellulose filter, detection line and nature controlling line are parallel to each other,
Gauge is from for 3~5mm, and wherein detection line is close to sample pad, and nature controlling line is away from sample pad;Bladder chalone C monoclonal resists in detection line
Body CysC2 coating concentration is 0.3~2mg/ml, dosage is 0.5~1.5 μ l coating liquid measure/cm films, sheep anti-mouse igg in nature controlling line
Antibody coating concentration is 0.3~2mg/ml, dosage is 0.5~1.5 μ l coating liquid measure/cm films;
It is described to get stuck including upper casing and lower casing, well and observation window are provided with upper casing, the well corresponds to detection
The sample pad of test strips, the observation window correspond to detection line and nature controlling line on nitrocellulose filter, and the upper casing and lower casing can
Detachable is spliced, and the bayonet of fixed test test strips is provided with lower casing, and test strip can be replaced as needed, card
Shell repeats utilization.
The test strip is prepared by the following method:
(1) activation of time-resolved fluorescence microballoon
After ultrasonication fluorescent microsphere 2min, after taking 200 μ l fluorescent microspheres with 5~15min of 14000rpm high speed centrifugations,
The MES solution that sediment 10~100mM, pH are 5.0~7.0 is washed to 1ml, ultrasonication 2min;Add 10~100 μ l
20~100mg/ml carbodiimides, mix 5~10min, add 20~100mg/ml N- hydroxies of 50~200 μ l
Succinimide, mixes 14000rpm 5~15min of high speed centrifugation after 10~20min, and precipitation is molten with the MES that pH is 5.0~7.0
Liquid is washed to 1ml;
(2) preparation of time-resolved fluorescence microballoon mark bladder chalone C monoclonal antibody CysC1
After fluorescent microsphere ultrasonication 2min after above-mentioned activation bladder chalone C is added according to 50~200 μ g/200 μ l
Monoclonal antibody CysC1, mix 1~3 it is small when, with containing 0.5%BSA, 10~50mM of 0.1% glycine, pH 7.5~
14000rpm 5~15min of high speed centrifugation after when 8.5Tris-HCl confining liquids closing 0.5~1 is small, with containing 1%Nacl, 0.5%
BSA, 0.5% sucrose, 10~50mM of 0.1%Tween20, pH7.5~8.5Tris-Hcl preserve liquid and wash and be resuspended to 200 μ
L is kept in dark place in 4 DEG C.
(3) preparation of sample pad
With sample pad treatment fluid (Tris-HCl of 20mM, pH8.0 containing 0.5%Nacl, 0.5%Tween, 0.1%BSA)
In the parallel even application two lines in side close to sample pad, dosage is 2~4 μ l liquid measures/cm sample pads, in sample pad close to bag
The bladder chalone C monoclonal antibody CysC1 that time-resolved fluorescence microballoon marks microballoon dilutions (are contained 0.5% by envelope side
The 20mM Tris-Hcl buffer solutions of BSA, 25% sucrose) dilution 8~30 times of even applications, one line, dosage for 2~4 μ l liquid measures/
Cm sample pads.It is placed in baking oven, 37 DEG C of drying are overnight.
(4) preparation of coated film
Coating buffer solution (the 10mM PBS buffer for containing 2.5% sucrose) is used respectively by bladder chalone C monoclonal antibody CysC2
Concentration is adjusted to 0.3~2mg/ml with sheep anti-mouse igg antibody, and dosage is coated with liquid measure/cm films for 0.5~1.5 μ l, respectively as inspection
In being coated with nitrocellulose filter, nature controlling line, detection line and microballoon line 3~7mm of interval, put parallel with nature controlling line stroke of survey line
In baking oven, 37 DEG C of drying are overnight.
(5) sequentially mutually pasting sample pad, coated film and blotting paper obtains test paper plate overlap joint on end liner, and cutting obtains
The test strips.
The test strip end liner size is 80*4mm, and sample pad size is 30*4mm, and coated film size is 25*4mm,
Water suction paper size is 28*4mm.
The utility model further relates to a kind of bladder chalone C time-resolved fluorescence detection kit, including:
(1) the bladder chalone C time-resolved fluorescence detection card described in;
(2) sample diluting liquid;
(3) SD card containing calibration curve.
The testing principle of this kit is double antibody sandwich method, bladder chalone C in detection human serum, blood plasma and whole blood sample
Content.Blood sample dilution containing bladder chalone C is added dropwise in sample application zone, is chromatographed through capillary action to sample pad, with when
Between resolved fluorometric microballoon mark bladder chalone C monoclonal antibody CysC1 combine, formed microballoon Antibody-antigen complex, chromatography extremely
Detection zone on nitrocellulose filter, reaction compound are detected the coated bladder chalone C monoclonal antibody CysC2 captures of survey line, and more
Remaining Fluorescent microsphere marker continues to chromatograph forward, is combined with being fixed on nature controlling line sheep anti mouse.After reaction, ultraviolet source is used
(365nm) to detection zone Scanning Detction, time-resolved fluorescence microballoon sends fluorescence (615nm) in detection line and nature controlling line, it declines
It is also longer to become the time.Using detection time is delayed, treat that abiogenous short life fluorescence (1~10ns) is all declined in sample substrate
After change, then the specificity fluorescent of rare earth element is measured, can thus exclude the interference of special background fluorescence completely.Pass through detection
The power and its ratio of line and nature controlling line fluorescence intensity, you can analyze the concentration of determinand in sample.
The sample diluting liquid composition is as follows:NaCl 0.5%, Tween 1%, solvent for 10~50mM, pH7.5~
8.5Tris-HCl buffer solutions, are sub-packed in centrifuge tube with 495 μ l/ pipes.This sample diluting liquid is used to chromatograph sample.
The SD card containing standard curve, is the quality-control product of passage time resolved fluorometric detection card measure various concentrations,
Using quality-control product concentration as abscissa, using fluorescence signal ratio as ordinate, standard curve is depicted as, writes and generates corresponding two
Dimension code information, which is stored in SD card, to be obtained.Corresponding 2 D code information on reagent card can be read by dry type fluorescence immunity analyzer
And measure respective concentration.
The beneficial effects of the utility model are mainly reflected in:The utility model detection card and kit being capable of accurate quantitative analysis inspections
The content of bladder chalone C, using time-resolved fluorescence microballoon detection technique, can avoid sample in survey human serum, blood plasma and whole blood sample
Microballoon is connected by this fluorescence interference itself, mark by covalent bond with antibody, and marked product is stablized, wide by (0.2 with detection range
~10mg/L), high sensitivity (detection limit 0.1mg/L), the features such as accuracy is high, detection is fast and convenient.
(4) illustrate
Fig. 1 is that the time-resolved fluorescence of the utility model quantitatively detects the knot of test strip in the kit of bladder chalone C
Structure schematic diagram;
Wherein:1st, sample pad;2nd, coated film;3rd, blotting paper;4th, microballoon line;5th, detection line;6th, nature controlling line;7th, end liner.
Fig. 2 is that the time-resolved fluorescence of the utility model quantitatively detects and the structure of card is detected in the kit of bladder chalone C shows
It is intended to;
Wherein:8th, Quick Response Code;9th, well;10th, observation window.
Fig. 3 is the kit standard curve prepared by the utility model embodiment 1.
Fig. 4 is that the kit prepared by the utility model embodiment 1 detects bladder chalone C kit with Roche immunoturbidimetry
Testing result correlation compares.
(5) embodiment
The utility model is described further with reference to specific embodiment, but the scope of protection of the utility model is simultaneously
It is not limited only to this:
Embodiment 1:Bladder chalone C time-resolved fluoroimmunoassay chromatographs the preparation of immue quantitative detection reagent box
Bladder chalone C time-resolved fluoroimmunoassay chromatographs immue quantitative detection reagent box, former using double antibody sandwich method immunochromatography
Manage, bladder chalone C in detection human serum, blood plasma and whole blood sample.By time-resolved fluorescence detection card, Sample dilution and containing fixed
The SD card composition of mark song line.
As shown in Figure 1, sequentially overlapping sample pad 1, coated film 2 and blotting paper 3 on end liner 7, sample pad 1 is sample application zone,
For drawing detection sample to be measured, equipped with microballoon line, coated film 2 is equipped with detection line and nature controlling line.
It is glimmering using the rare-earth europium of specific exciting light (365nm)/transmitting light (615nm) wavelength on microballoon line in sample pad 1
Light microballoon (diameter about 200nm, europium solid content 1%) mark bladder chalone C monoclonal antibody CysC1 (the μ l fluorescence of 100 μ g antibody/200
Microballoon), detection line coating bladder chalone C monoclonal antibody CysC2 (0.5mg/ml), the goat-anti that nature controlling line coating concentration is 1mg/ml
(wherein the buying of time-resolved fluorescence microballoon is from Bangs Lab companies of the U.S., and the buying of bladder chalone C monoclonal antibody is certainly for mouse IgG antibody
Guangzhou Growth hormone secretagogue bio tech ltd, sheep anti-mouse igg antibody come from Changsha Bo You bio tech ltd).Microballoon
Line dosage is that 4 μ l coatings liquid measure/cm sample pads, detection line and nature controlling line dosage are 1 μ l coating liquid measure/cm films.
In this embodiment, time-resolved fluorescence quantitatively detects the preparation of the kit of bladder chalone C and comprises the following steps:
1. the activation of time-resolved fluorescence microballoon
After ultrasonication fluorescent microsphere 2min, after taking 200 μ l fluorescent microspheres with 14000rpm high speed centrifugations 15min, sink
The MES solution that starch 100mM, pH are 6.0 is washed to 1ml, ultrasonication 2min;The 100mg/ml carbon two for adding 50 μ l is sub-
Amine, mixes 5min, adds the 100mg/ml N- hydroxy thiosuccinimides of 100 μ l, mixes 14000rpm high after 15min
Speed centrifugation 15min, precipitates and is washed with the MES solution that pH is 6.0 to 1ml.
2. the preparation of time-resolved fluorescence microballoon mark bladder chalone C monoclonal antibody CysC1
After fluorescent microsphere ultrasonication 2min after above-mentioned activation bladder chalone C Dan Ke is added according to 100 μ g/200 μ l
Grand antibody, mix 2 it is small when, with containing 0.5%BSA, 0.1% glycine 50mM, pH8.0Tris-Hcl confining liquid closing 1 it is small when
14000rpm high speed centrifugations 15min afterwards, with containing 1%Nacl, 0.5%BSA, 0.5% sucrose, 0.1%Tween20 50mM,
Buffer solution washes twice and is ultrasonically treated resuspension in the Tris-HCl preservation liquid of pH8.0 is kept in dark place to 200 μ l in 4 DEG C.
3. the preparation of sample pad
With sample pad treatment fluid (Tris-HCl of 20mM, pH8.0 containing 0.5%Nacl, 0.5%Tween, 0.1%BSA)
In the parallel even application two lines in side close to sample pad, dosage is 4 μ l liquid measures/cm sample pads, in sample pad close to coating
Film side by the bladder chalone C monoclonal antibody CysC1 that time-resolved fluorescence microballoon marks with microballoon dilution (containing 0.5%BSA,
The 20mM Tris-Hcl buffer solutions of 25% sucrose) dilution 15 times of even applications, one line (i.e. microballoon line), dosage be 4 μ l liquid measures/
Cm sample pads.It is placed in baking oven, 37 DEG C of drying are overnight.
4. the preparation of coated film
Coating buffer solution (the 10mM PBS buffer for containing 2.5% sucrose) is used respectively by bladder chalone C monoclonal antibody CysC2
Adjust concentration respectively with sheep anti-mouse igg antibody and be coated with liquid measure/cm films to 0.5mg/ml, 1mg/ml, dosage for 1 μ l, respectively as
Parallel with nature controlling line stroke of detection line on nitrocellulose filter in being coated with, nature controlling line, detection line interval 4mm, in humidity<
30%, dry in 37 DEG C of baking ovens of temperature 10 it is small when, envelope is spare;
5. sequentially sample pad (size 30*300mm, glass are pasted in mutually overlap joint ground on end liner (size 80*300mm)
Glass cellucotton material), coated film (size 25*300mm, nitrocellulose material) and blotting paper (size 28*300mm)
To test paper plate, the test strips of 4mm width are cut into as requested.
The utility model test strip, when in use, test strips be assembled in fastened by plastics upper casing and plastics lower casing and
Into plastic shell in, plastics upper casing is equipped with two perforates, well 9 and observation window 10, and well 9 corresponds to the detection
The sample pad 1 of the test strips of bladder chalone C, the bayonet of fixed test test strips is provided with plastics lower casing, and observation window 10 corresponds to
The detection line 5 and nature controlling line 6 of the test strips of the detection bladder chalone C, the test strips of the detection bladder chalone C can be outside the plastics
Taken out in shell.
In the utility model kit, further include Sample dilution, for the 50mM containing 0.5%NaCl, 1%Tween,
PH8.0Tris-Hcl buffer solutions, are sub-packed in centrifuge tube with 495 μ l/ pipes.Sample dilution is used for chromatography samples.
In the utility model kit, per SD card (with batch standard curve identical) of the box containing standard curve, pass through
The quality-control product of time-resolved fluorescence detection card measure various concentrations, using quality-control product concentration as abscissa, is made with fluorescence signal ratio
For ordinate, standard curve is depicted as, SD card is write and generates Quick Response Code, reagent can be read by dry type fluorescence immunity analyzer
Corresponding 8 information of Quick Response Code and respective concentration is measured on card.
Embodiment 2:Bladder chalone C time-resolved fluoroimmunoassay chromatographs the quantitative detection of immue quantitative detection reagent box
1. the drafting of standard curve
The time-resolved fluorescence detection card prepared by embodiment 1 adds various concentrations bladder chalone C antigen quality-control product
(totally 7 concentration, each concentration set three weights by 10mg/L, 7.5mg/L, 5mg/L, 2mg/L, 1mg/L, 0.5mg/L, 0.2mg/L
Multiple, bladder chalone C antigen is diluted to obtain the quality-control product mother liquor of 10mg/L with calf serum, then is diluted to different Quality Controls with calf serum
Product concentration), Sample dilution is added, after chromatographing 5min, passes through the AFS-1000 of Guangzhou Lan Bo bio tech ltd production
Type dry type fluorescence immunity analyzer reads C, T line fluorescence signal and C/T values, experimental result and analysis in table 1:
Standard curve is drawn with bladder chalone C quality-control product concentration and sample signal T/C average values, curve data is shown in Table 1, standard
Curve is as shown in Figure 3.Wherein R values are 0.9984, and concentration quantitative survey is carried out to contained bladder chalone C concentration in sample by the graticule
It is fixed.
2. time-resolved fluorescence test card performance test.
Minimum detectability:Replication is carried out with null value sample 20 times, calculates the average M and standard deviation SD of 20 results,
Add the test limit (M+2SD) of twice of standard deviation method for reporting with blank average, be as a result 0.08mg/L, meet sensitivity 0.1mg/
L。
The range of linearity:7 concentration values between 0.2~10mg/L are taken, three times, measured concentration is put down for each concentration duplicate measurements
Average carries out linear analysis with theoretical concentration, obtains linear equation y=1.0614x-0.1343, r=0.9977, shows this practicality
Novel agent box correlation in 0.2~10mg/L ranges of linearity is fine.
Precision:Take made time-resolved fluorescence in three batches of embodiments 1 quantitatively to detect bladder chalone C kit, detect respectively
7.5th, 1mg/L repeatability quality-control product, every batch of kit use repeatability quality-control product Parallel testing 10 times, 7.5mg/L concentration three batches batches
Interior CV is respectively 6.06%, 5.24%, 6.28%, and CV is respectively for CV in 7.65%, 1mg/L concentration three batches batches between three batches batches
7.06%th, 8.21%, 9.37%, CV is 9.57% between three batches batches, within 10%.
Accuracy:Selection concentration is detection sample for the Quality Control thing of 0.5mg/L, and it is 3 parts identical to be divided into volume, 2 parts wherein
The accuracy quality-control product of 7.5mg/L and 5mg/L is separately added into sample, 2 different recycling samples for adding concentration are made, calculate
The concentration of the determinand of addition.The solution without measured object of same amount is added in another sample, basic sample is made.To sample
3 replicate analysis of this progress, take its average to be calculated.The rate of recovery=(analysis sample concentration-basis sample concentration)/is calculated to add
Enter concentration × 100%.The rate of recovery that the rate of recovery for recycling sample 7.5mg/L is 99.73%, 5mg/L is 104.87%, average to return
Yield is 102.30%.Deviation is within 10%.
3. clinical sample detects
200 parts of the plasma sample of hospital's detection bladder chalone C is gathered, with the kit and Roche immunoturbidimetry of the utility model
Two methods of method detection bladder chalone C kit, which are detected, to be compared.In the utility model kit, 5 μ l of plasma sample are taken to add
Sample dilution, takes 80 μ l to be added in detection card well after mixing, have after chromatographing 5min by the blue vigorous biotechnology in Guangzhou
The AFS-1000 type dry types fluorescence immunity analyzer of limit company production reads concentration, and same sample is immunized using comparison system Roche
Turbidimetry detection bladder chalone C kit carries out Concentration Testing.Two methods testing result carries out linear analysis, as shown in figure 4, its
Correlation fine r=0.9812, P>0.05, mean relative deviation is less than 10%, as a result meets clinical analysis requirement, is suitable for
Clinical detection.
Claims (3)
1. a kind of bladder chalone C time-resolved fluorescence detection card, including get stuck and be arranged on the interior test strip that gets stuck:
The test strip is by the end liner and glass fiber sample pad being pasted onto on end liner, nitrocellulose filter and blotting paper
Composition, the sample pad, nitrocellulose filter and blotting paper, which sequentially overlap, to be pasted onto on end liner;
The sample pad is provided with microballoon line, is coated with the bladder chalone C monoclonal antibody of time-resolved fluorescence microballoon mark
CysC1, the rare earth doped europium element of time-resolved fluorescence microballoon, particle diameter is 100nm~300nm;
Detection line and nature controlling line are disposed with the nitrocellulose filter, detection line and nature controlling line are parallel to each other, spacer
From for 3~5mm, wherein detection line is close to sample pad, and nature controlling line is away from sample pad;
It is described to get stuck including upper casing and lower casing, well and observation window are provided with upper casing, the well corresponds to Test paper
The sample pad of bar, the observation window correspond to detection line and nature controlling line on nitrocellulose filter, and the upper casing and lower casing are detachable
Formula is spliced, and the bayonet of fixed test test strips is provided with lower casing.
2. bladder chalone C time-resolved fluorescence detection card as claimed in claim 1, it is characterised in that the test strip bottom
Lining size is 80*4mm, and sample pad size is 30*4mm, and coated film size is 25*4mm, and water suction paper size is 28*4mm.
3. a kind of bladder chalone C time-resolved fluorescence detection kit, including:
(1) the bladder chalone C time-resolved fluorescence detection card described in claim 1 or 2;
(2) sample diluting liquid;
(3) SD card containing calibration curve.
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