CN208752087U - A kind of fluorescence immune chromatography detection kit of three kinds of cardiac markers of quantitative detection - Google Patents
A kind of fluorescence immune chromatography detection kit of three kinds of cardiac markers of quantitative detection Download PDFInfo
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Abstract
Disclose a kind of compound quantitative detection hs-CRP (Hs-CRP), myeloperoxidase (MPO), lipoprotein phospholipase A2 (Lp-PLA2) fluorescence immune chromatography detection kit, including desiccant, hermetic bag and detection card.The detection card includes triplet buckle and reagent strip；The reagent strip includes sample pad, bonding pad, blood filter membrane, nitrocellulose filter, blotting paper and PVC bottom plate, and the bonding pad is coated with hsCRP antibody, MPO antibody and Lp-PLA2 antibody.This method has reached minimum, detection efficiency height with high, quick, easy, reproducible, the easy automation of special, sensitive, yield, false positive, and testing cost is low to wait outstanding advantages, there is important Practical significance.
Description
Technical field
The utility model relates to clinical medical inspection fields, specifically, being related to a kind of super sensitive C-reactive of joint quantitative detection
Albumen, myeloperoxidase and lipoprotein phospholipase A2 fluorescence immune chromatography detection kit.
Background technique
Cardiovascular disease is clinically common Severe acute disease, and case fatality rate is high, the state of an illness is dangerous, seriously threatens the mankind's
Health and life.Data show that cardiovascular disease basic pathology variation be atherosclerosis, in recent years researches show that inflammation
Disease reaction is that the basis of atherosclerosis and cardiovascular disease, especially chronic inflammation are considered to increase arterial injury disease
Risk makes atherosclerotic plaque be easy to rupture, and has played key effect in the formation of atherosclerosis.Arterial injury is
A kind of chronic inflammatory reaction result of whole body cardiovascular system.But since arterial injury early stage lacks apparent clinical manifestation, institute
Many places are in the middle and advanced stage of disease when making a definite diagnosis, cause patient poor prognosis and excessive health care costs.Then effective prison is found
Survey early stage arterial disease, the cardiovascular and cerebrovascular marker of the serious cardiovascular and cerebrovascular diseases risk of prediction has important clinical meaning.
Super quick c reactive protein (Hs-CRP) is the Sensitive mark of inflammatory disorders, can accurately detect low concentration C reaction egg
It is white.Clinical test shows serum Hs-CRP level as the coronary heart disease state of an illness is in gradual raising trend.Hs-CRP is reflection inflammatory
Sensitive, the accurate index of process, slight inflammation can cause Hs-CRP to increase in vivo, and epidemiological study, which is shown, increases
Hs-CRP can indicate the CHD in 10 years occur.Lipoprotein phospholipase A2 (Lp-PLA2) is point of calcium ion dependent/non-dependent
The secreted protein that son amount is 45KDa.Lp-PLA2 is generated by macrophage, is the marker of blood vessel endothelium inflammation, not by other
Diseases associated with inflammation influences.It can be with the generation of specific detection early stage vascular inflammation.Multinomial clinical experimental study shows basic Lp-
The horizontal junior of the more basic Lp-PLA2 of ratio that cardiovascular event occurs after the horizontal the higher person of PLA2 significantly increases.Marrow peroxide
Compound enzyme (MPO) is a kind of Neprilysin body enzyme, is generated, is participated in by the neutrophil leucocyte, monocyte and macrophage that activate
Various inflammatory reactions and oxidative stress process in human body.MPO increases prompt patient, and there may be coronary inflammations, but blood vessel is not
It is completely plugged, do not cause myocardial necrosis, can be used for predicting a possibility that early stage adverse cardiac events occur.This is practical new
A kind of compound quantitative detection hs-CRP of type, myeloperoxidase and the detection of lipoprotein phospholipase A2 fluorescence immune chromatography
The above triplicity is passed through joint-detection serum high-sensitive c reactive protein, myeloperoxidase and lipoprotein phosphatide by kit
Enzyme A2 is horizontal, can more accurately and effectively predict the possibility of the serious cardiovascular and cerebrovascular diseases risk of patient, realizes serious cardiovascular thing
Part early diagnosis, risk profile and seriousness assessment.
Chinese invention patent CN201010102136.5 discloses one kind albumen containing vascular peroxidase and antibody ELISA
Kit, this method judge the severity of the coronary heart disease state of an illness, single index by single vascular peroxidase concentration
Diagnosis accuracy is not high, and ELISA method accuracy of judgement degree is not high, false positive easily occurs, and time-consuming for detection, test operation is multiple
It is miscellaneous.Chinese invention patent CN201410623721.8 discloses a kind of blood serum designated object Samd3 albumen for diagnosis of coronary heart disease
ELISA kit equally applies ELISA method, and accuracy is not high, false positive easily occurs, and time-consuming for detection, test operation is multiple
It is miscellaneous, major cardiovascular events can not accurately be early diagnosed, the assessment of risk profile and seriousness.
In recent years, cardiovascular event has become China human mortality and dies of illness first of reason, therefore EARLY RECOGNITION diagnosis and to it
Carrying out risk stratification has important clinical meaning.Therefore, a kind of pair of cardiac and cerebral vascular diseases diagnosis and risk stratification point are invented
It is necessary to analyse the high detection kit of accuracy.The utility model use in conjunction angiocarpy marker Hs-CRP, MPO, Lp-
The fluorescence immune chromatography method of PLA2 is predicted and is diagnosed to cardiovascular disease, compared with traditional index, Hs-CRP, MPO, Lp-
The use in conjunction of PLA2 accuracy in terms of major cardiovascular events early diagnosis and risk profile is higher, with the obvious advantage.And this
Utility model uses fluorescence immune chromatography method, have special, sensitive, yield it is high, quick, easy, reproducible, easy automate,
Positive rate has reached the outstanding advantages such as minimum, detection efficiency are high, testing cost is low up to 90% or more, false positive, there is weight
The Practical significance wanted.
Summary of the invention
Utility model aims to solve the deficiencies in the prior art, provide that a kind of high sensitivity, accuracy be strong, operation
Easy joint quantitative detection hs-CRP, myeloperoxidase and the detection examination of lipoprotein phospholipase A2 fluorescence immune chromatography
Agent box.
Realizing the technical solution of the utility model aim is: a kind of joint quantitative detection hs-CRP, marrow peroxidating
Enzyme and lipoprotein phospholipase A2 fluorescence immune chromatography detection kit, which is characterized in that the detection kit includes detection
Card, desiccant and hermetic bag, the detection card includes buckle and reagent strip；The reagent strip includes sample pad, bonding pad, nitric acid
Cellulose membrane, blotting paper and PVC bottom plate, the bonding pad are coated with hsCRP antibody, MPO antibody and Lp-PLA2 antibody；It is described
HsCRP antibody is one or more combinations of pairs of hsCRP monoclonal antibody and hsCRP polyclonal antibody；The MPO antibody is
One or more combinations of pairs of MPO monoclonal antibody and MPO polyclonal antibody；The Lp-PLA2 antibody is Lp-PLA2 Dan Ke
One or more combinations of pairs of grand antibody and Lp-PLA2 polyclonal antibody.
Further, the reagent strip further includes blood filter membrane.
Further, the preparation method of the detection kit includes the following steps:
(1) preparation of sample pad: sample pad selects glass fibre element film, with salt ions such as the dissolution such as buffer NaCl, BSA
And albumen, a small amount of surfactant is then added, adjusts pH to 7-8, is uniformly laid on glass fibre by glass fibre water absorption
On plain film, it is placed in 37 DEG C of dryings 8 hours.
(2) it is (anti-that the sample pad handled well uniformly the preparation of bonding pad: is spread to the fluorescent microsphere labelled antibody diluted
Hs-CRP antibody, anti-myeloperoxidase antibody or anti-grease protein, phospholipid enzyme A2 antibody) solution, after vacuum freeze drying
Sealing, room temperature preservation.
(3) blood filter membrane the preparation of blood filter membrane: is cut into 10cm × 4mm every.
(4) processing of nitrocellulose filter:
A. prepared by detection line: goat-anti labelled antibody is diluted to the concentration of 1.0mg/mL, 0.8ul/cm with dilution coating buffer
Scribing rates cross on nitrocellulose filter and obtain detection line, nitrocellulose filter is placed in 37 DEG C of dryings after scribing line
8 hours to obtain the final product；
B. prepared by nature controlling line: sheep anti-mouse igg antibody is diluted to the dense of 1.0mg/mL with the PB buffer of 50mM/LpH7.2
Degree, the scribing rates of 0.8ul/cm cross on nitrocellulose filter and obtain nature controlling line, by nitrocellulose filter after scribing line
It is placed in 37 DEG C of dryings 8 hours to obtain the final product.
(5) blotting paper the preparation of blotting paper: is cut into 30cm × 2.7cm every.
(6) it assembles: by the above-mentioned sample pad handled well, microballoon label bonding pad, blood filter membrane, nitrocellulose filter and water suction
Paper is successively pasted on PVC bottom plate, and the test strips of wide 4mm are cut into after posting, and is placed in triplet buckle, and drying is added
Agent encapsulates to obtain the final product.
Further, the buffer of the preparation of the sample pad is selected from one of Tris, PBS or glycine, described
Surfactant is selected from one of Tween20 or TritonX-100.
Further, the fluorescent microsphere of the preparation of the bonding pad is selected from fluorescein cy5, fluorescein cy7, polyphenyl containing europium
One of ethylene microballoon, 2- methoxyl group fluorescence enhancement element, rhodamine, phycoerythrin.
The good effect that the utility model has: the fluorescence immune chromatography kit of the utility model is in blood sample
Hs-CRP, MPO, Lp-PLA2 carry out quantitative detection, and compared with traditional index, the use in conjunction of Hs-CRP, MPO, Lp-PLA2 exist
Accuracy is higher in terms of major cardiovascular events early diagnosis and risk profile, with the obvious advantage.Another the utility model uses fluorescence
Immunochromatographic method is compared with the traditional method, and has special, sensitive, high, quick, easy, reproducible, the easy automation of yield, sun
Outstanding advantages of property recall rate has reached minimum up to 90% or more, false positive, there is important Practical significance.
Detailed description of the invention
Fig. 1 is the schematic diagram of the utility model reagent strip structure.
Fig. 2 is combination buckle structural schematic diagram.
Fig. 3 is triplet buckle structural schematic diagram.
Fig. 4 is hs-CRP examination criteria working curve diagram.
Fig. 5 is myeloperoxidase examination criteria working curve diagram.
Fig. 6 is lipoprotein phospholipase A2 examination criteria working curve diagram.
Fig. 7 is hs-CRP linear criterion working curve diagram.
Fig. 8 is myeloperoxidase linear criterion working curve diagram.
Fig. 9 is lipoprotein phospholipase A2 linear criterion working curve diagram.
Specific embodiment
The utility model is further described in following embodiment combination attached drawings, but the utility model be not limited to
Lower explanation.
As shown in Figure 1, Figure 2, Figure 3 shows, the utility model detection kit is in addition to the desiccant and hermetic bag, primary structure
Detection card includes buckle 7 or buckle 10 and reagent strip 1, and wherein reagent strip 1 includes sample pad 5, bonding pad 3, nitrocellulose filter
2, blotting paper 4 and PVC bottom plate 6；Buckle includes combination buckle 7 and three buckles 10, and buckle includes well 8 and observation window 9,
Reagent strip 1 is placed in buckle 1 or buckle 10.
Embodiment 1:
(1) preparation of sample pad
Sample pad selects glass fibre element film, dissolves salt ions and the albumen such as Nacl, BSA with buffer Tris etc., then
A small amount of surfactant Tween20 is added, adjusts pH to 7-8, is uniformly laid on glass by glass water absorption, is placed in 37 DEG C and does
Dry 8 hours to obtain the final product.
(2) preparation of bonding pad
The monoclonal antibody that fluorescent microsphere is marked, is diluted with microballoon dilution, is uniformly layered on the sample handled well
It on pad, is sealed after vacuum freeze drying, room temperature preservation, wherein fluorescent microsphere labelled antibody object preparation process is as follows:
A. the coupling of fluorescent microsphere and anti-hs-CRP monoclonal antibody
The fluorescent microsphere that particle size is 300nm is first activated with EDC and NHS activator and stabilizer, has been activated
After cleaned, remove activator, then mix again with the hs-CRP monoclonal antibody of 250ug/mL, room temperature reaction
2h, the extra antibody of cleaning removal supernatant, then carries out 1 hour of closing with 2% casein solution, then delays after completion of the reaction
Fliud flushing is cleaned several times, is finally saved liquid with microballoon and is saved, obtains the hs-CRP monoclonal antibody of fluorescent microsphere label,
The launch wavelength of fluorescent microsphere is 615nm.
B. the coupling of fluorescent microsphere and anti-myeloperoxidase monoclonal antibody
The fluorescent microsphere that particle size is 300nm is first activated with EDC and NHS activator and stabilizer, has been activated
After cleaned, remove activator, then mix again with the myeloperoxidase protein monoclonal antibody of 250ug/mL, room temperature reaction
2h, the extra antibody of cleaning removal supernatant, then carries out 1 hour of closing with 2% casein solution, then delays after completion of the reaction
Fliud flushing is cleaned several times, is finally saved liquid with microballoon and is saved, obtains the myeloperoxidase monoclonal antibody of fluorescent microsphere label, fluorescence
The launch wavelength of microballoon is 615nm.
C. the coupling of fluorescent microsphere and anti-grease protein, phospholipid enzyme A2 monoclonal antibody
The fluorescent microsphere that particle size is 300nm is first activated with EDC and NHS activator and stabilizer, has been activated
After cleaned, remove activator, then mix again with the lipoprotein phospholipase A2 monoclonal antibody of 250ug/mL, room temperature reaction
2h, the extra antibody of cleaning removal supernatant, then carries out 1 hour of closing with 2% casein solution, then delays after completion of the reaction
Fliud flushing is cleaned several times, is finally saved liquid with microballoon and is saved, obtains the lipoprotein phospholipase A2 monoclonal antibody of fluorescent microsphere label,
The launch wavelength of fluorescent microsphere is 615nm.
(3) blood filter membrane the preparation of blood filter membrane: is cut into 10cm × 4mm every.
(4) processing of nitrocellulose filter
A. the processing of hs-CRP nitrocellulose filter: the sheep anti-mouse igg antibody PB of 50mM/L pH7.2 is delayed
Fliud flushing is diluted to the concentration of 1.0mg/mL, and the scribing rates of 0.8ul/cm cross to obtain Quality Control in NC film upper end C line labeling position
Line；Goat-anti hs-CRP polyclonal antibody is also diluted to dilution coating buffer to the concentration of 1.0mg/mL, 0.8ul/cm
Scribing rates on NC film T line labeling position cross to obtain quantitative detection line, 5mm is divided between detection line and nature controlling line, cross
It finishes and is placed on 37 DEG C of dryings 8 hours to obtain the final product.
B. the processing of myeloperoxidase test strips NC film: by the PB buffer of sheep anti-mouse igg antibody 50mM/L pH7.2
It is diluted to the concentration of 1.0mg/mL, the scribing rates of 0.8ul/cm cross to obtain nature controlling line in NC film upper end C line labeling position,
Goat-anti people's myeloperoxidase polyclonal antibody is also diluted to the concentration of 1.0mg/mL with dilution coating buffer, 0.8ul/cm's draws
Line rate T line labeling position on NC film crosses to obtain quantitative detection line, 5mm is divided between detection line and nature controlling line, scribing line finishes
It is placed on 37 DEG C of dryings 8 hours to obtain the final product.
C. the processing of lipoprotein phospholipase A2 test strips NC film: the sheep anti-mouse igg antibody PB of 50mM/L pH7.2 is delayed
Fliud flushing is diluted to the concentration of 1.0mg/mL, and the scribing rates of 0.8ul/cm cross to obtain Quality Control in NC film upper end C line labeling position
Goat-anti human lipoprotein phospholipase A2 polyclonal antibody is diluted to the concentration of 1.0mg/mL, 0.8ul/cm by line with dilution coating buffer
Scribing rates on NC film T line labeling position cross to obtain quantitative detection line, 5mm is divided between detection line and nature controlling line, cross
It finishes and is placed on 37 DEG C of dryings 8 hours to obtain the final product.
(5) blotting paper the preparation of blotting paper: is cut into 30cm × 2.7cm every.
(6) it assembles: by the above-mentioned sample pad handled well, microballoon label bonding pad, blood filter membrane, nitrocellulose filter and water suction
Pad is successively pasted on PVC bottom plate, is cut into the test strips of wide 4mm after posting with cutting machine, is placed in triplet buckle, adds
Enter desiccant encapsulation, it is glimmering to be configured to hs-CRP, myeloperoxidase and lipoprotein phospholipase A2 jointly with dilution buffer
Light immunochromatographytest test kit.
Embodiment 2:
(1) preparation of sample pad
Sample pad selects glass fibre element film, with dissolution salt ions and the albumen such as Nacl, BSA such as buffer PBS, then plus
Enter a small amount of surfactant TritonX-100, adjusts pH to 7-8, be uniformly laid on glass by glass water absorption, be placed in 37 DEG C
Dry 8 hours to obtain the final product.
(2) preparation of bonding pad
The monoclonal antibody that fluorescent microsphere is marked, is diluted with microballoon dilution, is uniformly layered on the sample handled well
It on pad, is sealed after vacuum freeze drying, room temperature preservation, wherein fluorescent microsphere labelled antibody object preparation process is as follows:
A. the coupling of fluorescent microsphere and anti-hs-CRP monoclonal antibody
The fluorescent microsphere that particle size is 300nm is first activated with EDC and NHS activator and stabilizer, has been activated
After cleaned, remove activator, then mix again with the hs-CRP monoclonal antibody of 250ug/mL, room temperature reaction
2h, the extra antibody of cleaning removal supernatant, then carries out 1 hour of closing with 2% casein solution, then delays after completion of the reaction
Fliud flushing is cleaned several times, is finally saved liquid with microballoon and is saved, obtains the hs-CRP monoclonal antibody of fluorescent microsphere label,
The launch wavelength of fluorescent microsphere is 615nm.
B. the coupling of fluorescent microsphere and anti-myeloperoxidase monoclonal antibody
The fluorescent microsphere that particle size is 300nm is first activated with EDC and NHS activator and stabilizer, has been activated
After cleaned, remove activator, then mix again with the myeloperoxidase protein monoclonal antibody of 250ug/mL, room temperature reaction
2h, the extra antibody of cleaning removal supernatant, then carries out 1 hour of closing with 2% casein solution, then delays after completion of the reaction
Fliud flushing is cleaned several times, is finally saved liquid with microballoon and is saved, obtains the myeloperoxidase monoclonal antibody of fluorescent microsphere label, fluorescence
The launch wavelength of microballoon is 615nm.
C. the coupling of fluorescent microsphere and anti-grease protein, phospholipid enzyme A2 monoclonal antibody
The fluorescent microsphere that particle size is 300nm is first activated with EDC and NHS activator and stabilizer, has been activated
After cleaned, remove activator, then mix again with the lipoprotein phospholipase A2 monoclonal antibody of 250ug/mL, room temperature reaction
2h, the extra antibody of cleaning removal supernatant, then carries out 1 hour of closing with 2% casein solution, then delays after completion of the reaction
Fliud flushing is cleaned several times, is finally saved liquid with microballoon and is saved, obtains the lipoprotein phospholipase A2 monoclonal antibody of fluorescent microsphere label,
The launch wavelength of fluorescent microsphere is 615nm.
(3) blood filter membrane the preparation of blood filter membrane: is cut into 10cm × 4mm every.
(4) processing of nitrocellulose filter
A. the processing of hs-CRP nitrocellulose filter: the sheep anti-mouse igg antibody PB of 50mM/L pH7.2 is delayed
Fliud flushing is diluted to the concentration of 1.0mg/mL, and the scribing rates of 0.8ul/cm cross to obtain Quality Control in NC film upper end C line labeling position
Line；Goat-anti hs-CRP polyclonal antibody is also diluted to dilution coating buffer to the concentration of 1.0mg/mL, 0.8ul/cm
Scribing rates on NC film T line labeling position cross to obtain quantitative detection line, 5mm is divided between detection line and nature controlling line, cross
It finishes and is placed on 37 DEG C of dryings 8 hours to obtain the final product.
B. the processing of myeloperoxidase test strips NC film: by the PB buffer of sheep anti-mouse igg antibody 50mM/L pH7.2
It is diluted to the concentration of 1.0mg/mL, the scribing rates of 0.8ul/cm cross to obtain nature controlling line in NC film upper end C line labeling position,
Goat-anti people's myeloperoxidase polyclonal antibody is also diluted to the concentration of 1.0mg/mL with dilution coating buffer, 0.8ul/cm's draws
Line rate T line labeling position on NC film crosses to obtain quantitative detection line, 5mm is divided between detection line and nature controlling line, scribing line finishes
It is placed on 37 DEG C of dryings 8 hours to obtain the final product.
C. the processing of lipoprotein phospholipase A2 test strips NC film: the sheep anti-mouse igg antibody PB of 50mM/L pH7.2 is delayed
Fliud flushing is diluted to the concentration of 1.0mg/mL, and the scribing rates of 0.8ul/cm cross to obtain Quality Control in NC film upper end C line labeling position
Goat-anti human lipoprotein phospholipase A2 polyclonal antibody is diluted to the concentration of 1.0mg/mL, 0.8ul/cm by line with dilution coating buffer
Scribing rates on NC film T line labeling position cross to obtain quantitative detection line, 5mm is divided between detection line and nature controlling line, cross
It finishes and is placed on 37 DEG C of dryings 8 hours to obtain the final product.
(5) blotting paper the preparation of blotting paper: is cut into 30cm × 2.7cm every.
(6) it assembles: by the above-mentioned sample pad handled well, microballoon label bonding pad, blood filter membrane, nitrocellulose filter and water suction
Pad is successively pasted on PVC bottom plate, is cut into the test strips of wide 4mm after posting with cutting machine, is placed in triplet buckle, adds
Enter desiccant encapsulation, it is glimmering to be configured to hs-CRP, myeloperoxidase and lipoprotein phospholipase A2 jointly with dilution buffer
Light immunochromatographytest test kit.
Embodiment 3:
The detection of hs-CRP, myeloperoxidase and lipoprotein phospholipase A2 fluorescence immune chromatography detection kit.
(1) standard curve is drawn
Various concentration hs-CRP, marrow peroxide are separately added on the kit sample pad prepared by embodiment 1
Change enzyme, lipoprotein phospholipase A2 antigen standard (respectively take 7 various concentrations, hs-CRP antigen standard is respectively 0,
0.5,5,10,40,100,150ng/mL, myeloperoxidase antigen standard is respectively 0,10,50,100,200,500,
1000ng/mL, lipoprotein phospholipase A2 antigen standard are respectively 0,50,100,200,500,1000 and 1500ng/mL, each
Concentration sets 3 repetitions), after 15 minutes, be placed in fluorescent quantitation immunochromatography instrument and obtain fluorescence signal intensity, according to detection line with
Nature controlling line fluorescence signal ratio, the concentration of institute's sample product is sought by standard curve.Experimental result and analysis are shown in Table 1~3:
1 hs-CRP standard items testing result of table
2 myeloperoxidase standard items testing result of table
3 lipoprotein phospholipase A2 standard items testing result of table
Respectively with hs-CRP, myeloperoxidase, lipoprotein phospholipase A2 antigen concentration of standard solution and measurement letter
Number average of relatives value draws standard curve, as shown in attached drawing 1~3.
(2) range of linearity is detected
The kit prepared using the utility model embodiment 1 does the linear range test of detection.
Hs-CRP standard items are taken, are diluted to 6 concentration, hs-CRP concentration model respectively with physiological saline
Enclosing is 0.5ng/mL-150ng/mL, each concentration replication 3 times, and the average value for measuring concentration and theoretical concentration are carried out line
Property regression analysis, calculate regression equation y=0.9957X+0.4789, correlation coefficient r=0.999 shows that this kit exists
Good relationship in the 0.5ng/mL-150ng/mL range of linearity (see attached drawing 4).
Myeloperoxidase standard items are taken, with normal saline dilution at 6 concentration, concentration range is 10-1000ng/mL,
The average value for measuring concentration and theoretical concentration are carried out linear regression analysis, calculate recurrence side by each concentration replication 3 times
Journey y=0.9986X+5.2168, correlation coefficient r=0.999 show this kit phase in the 10-1000ng/mL range of linearity
Closing property is preferably (see attached drawing 5).
Lipoprotein phospholipase A2 standard items are taken, with normal saline dilution at 6 concentration, concentration range is 50-1500ng/
The average value for measuring concentration and theoretical concentration are carried out linear regression analysis, calculate and return by mL, each concentration replication 3 times
Equation y=1.0346X-9.3357, correlation coefficient r=0.999 show this kit phase in the 50-1500ng/mL range of linearity
Closing property is preferably (see attached drawing 6).
(3) sensitivity (minimum detection limit)
Using human plasma as blank sample, it is measured, is repeated 20 times with test strips prepared by the utility model embodiment 1,
Calculating hs-CRP result mean value is 0.0208, standard deviation 0.0024, adds twice of standard deviation report according to blank mean value
It is 0.025 that method, which calculates fluorescence immunoassay quantitative analysis instrument measurement degree variable quantity, substitutes into standard curve and obtains minimum detectability
For 0.12ng/mL；Calculating myeloperoxidase result mean value is 0.0306, and standard deviation 0.0035 adds twice according to blank mean value
It is 0.0375 that standard deviation method for reporting, which calculates fluorescence immunoassay quantitative analysis instrument measurement degree variable quantity, substitutes into standard curve and obtains
Minimum detectability is 5.13ng/mL；Calculating lipoprotein phospholipase A2 result mean value is 0.015, standard deviation 0.0096, according to sky
It is 0.0342 that white mean value, which adds twice of standard deviation method for reporting to calculate fluorescence immunoassay quantitative analysis instrument measurement degree variable quantity, substitutes into mark
It is 20.5ng/mL that minimum detectability is obtained in directrix curve.
(4) repeatability and accuracy
The hs-CRP standard solution for preparing 1.0ng/mL and 10ng/mL, using the utility model embodiment 1
The kit of preparation is measured, and each concentration is distinguished replication 5 times, calculates separately measurement mean value and standard deviation.Calculate variation
Coefficient carries out repeated investigation, and the coefficient of variation is respectively 5.6% and 0.9% as the result is shown；With (1- mean value/standard value) ×
100%, which calculates relative deviation, carries out accuracy investigation, and relative deviation is respectively 4% and 6%.
The myeloperoxidase standard solution for preparing 50ng/mL and 200ng/mL is prepared using the utility model embodiment 1
Kit be measured, each concentration is distinguished replication 5 times, and measurement mean value and standard deviation are calculated separately.Calculate the coefficient of variation
Repeated investigation is carried out, the coefficient of variation is respectively 9.5% and 6.3% as the result is shown；In terms of (1- mean value/standard value) × 100%
It calculates relative deviation and carries out accuracy investigation, relative deviation is respectively 8.3% and 4.6%.
The lipoprotein phospholipase A2 standard solution for preparing 100ng/mL and 500ng/mL, using the utility model embodiment
The kit of 1 preparation is measured, and each concentration is distinguished replication 5 times, calculates separately measurement mean value and standard deviation.Calculate variation
Coefficient carries out repeated investigation, and the coefficient of variation is respectively 10.6% and 3.6% as the result is shown；With (1- mean value/standard value) ×
100%, which calculates relative deviation, carries out accuracy investigation, and relative deviation is respectively 8.6% and 10.6%.
Claims (2)
1. a kind of fluorescence immune chromatography detection kit of three kinds of cardiac markers of quantitative detection, which is characterized in that the detection
Kit includes detection card, desiccant and hermetic bag, and the detection card includes buckle and reagent strip；The reagent strip includes sample
Pad, bonding pad, nitrocellulose filter, blotting paper and PVC bottom plate, the bonding pad are coated with hsCRP antibody, MPO antibody and Lp-
PLA2 antibody；The hsCRP antibody is one or more combinations of pairs of hsCRP monoclonal antibody and hsCRP polyclonal antibody；
The MPO antibody is one or more combinations of pairs of MPO monoclonal antibody and MPO polyclonal antibody；The Lp-PLA2 antibody
For one or more combinations of pairs of Lp-PLA2 monoclonal antibody and Lp-PLA2 polyclonal antibody.
2. detection kit according to claim 1, which is characterized in that the reagent strip further includes blood filter membrane.
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