Immunity chromatography detection test paper and kit
Technical field
The present invention relates to immuno-chromatographic assay technology field, more particularly, to immunity chromatography detection test paper and kit.
Background technology
Colloidal gold solution refers to aurosol of the dispersed phase particles diameter between l -150nm, belongs to multiphase heterogeneous body
System, color is in salmon pink to aubergine.Colloidal gold can be used as probe to carry out cell surface and intracellular polysaccharide, protein, more
The large biological molecules such as peptide, antigen, hormone, nucleic acid are accurately positioned, and can be used for daily immunodiagnosis.Since colloidal gold is made
Have many good qualities for label, therefore from having obtained rapid hair in many research fields at home and abroad since the technology comes out
Exhibition.It has been widely used in Electronic Speculum, flow cytometer, Western blotting, protein staining, the field such as manufacture of external diagnosis reagent.Mesh
The preceding application in medical test is mainly immunochromatographic method.
Gold-marking immunity chromatographic technique (gold-labeled immunochromatography assay, GICA) is a kind of
The solid phase labelling that the multiple technologies such as colloidal gold-labeled method, immunoassay technology and Chromatographic techniques are combined together is immunized
Detection technique.Because its is easy to operate, quick, and there is very high specificity and sensitiveness, in recent years in clinical diagnosis, bedside
The application development of context of detection is rapid, is widely applied in body fluid detection.
Gold-marking immunity chromatography detecting test paper application colloidal gold technique and antibody, antigentic specificity rule are detected,
Work as test solution(Different according to the project of inspection, sample can use patients serum, saliva or urine)When instilling test paper, test solution passes through capillary
Effect, colloidal gold is taken on the p-wire for being adsorbed with antibody, if the antigen in test solution is corresponding with this antibody, is produced special
Property combine, colloidal gold is trapped on p-wire, in particular color.Using with light source and photodetector to Test paper into
The instant interpretation of row, can realize and target checking matter is quantitatively detected.
Conventional technique scheme, forms irradiation light on golden immunity chromatography detection test paper surface by LED light source, uses photoelectricity
Diode array receives the scattering light of golden immunity chromatography detection test paper, by the quantitative measurment to nanogold particle, realizes to mesh
Mark the Quantitative detection of gold label test strip.But shadow caused by system unit aging is not accounted in conventional technique scheme
Ring, in order to overcome the drawbacks described above of the prior art, 201210185414.7 patent of invention proposes a kind of gold-marking immunity chromatography
The performance correction method of ELISA test strip system, the data message of colloidal-gold strip colorimetric card when being dispatched from the factory by detecting system, according to this
Data message is established and the corresponding optimal response curve of detectable substance concentration;In the specific time cycle, colorimetric card is detected again
Corresponding data message, by it compared with calibration standard, so as to calculate correction factor, passes through the correction factor amendment system
The result for components aging of uniting.But also there are following defect for the technical solution of the invention:On the one hand, it needs to be spaced specific week
Phase can be modified, can not real-time judge whether need to be modified;On the other hand, it is only value revision that it, which is corrected,
Without considering specific physical process, the result is that nonlinear, accuracy deficiency.
The content of the invention
The present invention provides improved technical solution, can solve the above problem of the prior art, and improves modified
Accuracy.
As one aspect of the present invention, there is provided a kind of immunity chromatography detection test paper, including detection zone and calibration
Region;The detection zone sets detection line, when, there are during target detection thing, nano gold mark composition granule therein is sent out in sample
Raw immunochromatography reaction is gathered in detection line;The calibration region sets calibration color block, and the calibration color lump is positioned at described
The color range of nano gold mark composition granule, by for it is described calibration color lump detection be modified for testing result and
Determine the need for further correcting.
Preferably, the calibration color lump includes identical the first color lump and the second color lump of color, the first color lump table
Face covers the photic zone of specific light transmittance, and the second color lump surface does not cover photic zone.
Preferably, the euphotic light transmittance is 70%~90%.
Preferably, the euphotic light transmittance is 80%.
Preferably, first color lump and the second color are irradiated by detecting light source and standard sources respectively before detection
Block, according to photodetector in response to determining that the amendment of detection light source.
Preferably, the amendment includes the following steps:1st, basisCalculate correction factor k and b, wherein LMark 1
The signal strength that photodetector detects during to irradiate the first color lump using standard sources, LMark 2To use standard sources irradiation the
The signal strength that photodetector detects during two color lumps;LInspection 1Photodetector is examined during to irradiate the first color lump using detection light source
The signal strength of survey, LInspection 2The signal strength that photodetector detects during to irradiate the second color lump using detection light source;When the 2nd, detecting
By detecting light source irradiating and detecting region, the signal strength that photodetector detects is modified to LRepair=kLSurvey+b。
Preferably, the 3rd color lump and the 4th color lump are further included, the 3rd color lump and the 4th color lump are respectively difference
Color, wherein lower limit of the 3rd color lump color close to the color range of the nano gold mark composition granule, the 4th color lump color are leaned on
The upper limit of the color range of the nearly nano gold mark composition granule.
Preferably, by optimal response curve, the expectation signal strength of the 3rd color lump and the 4th color lump, L are calculatedPhase 3=
k1X1+b1And LPhase 4=k1X2+b1, wherein k1 and b1 are optimal response slope of a curve and intercept, and X1 and X2 are respectively the 3rd color lump
With the color range gradient magnitude of the 4th color lump;The signal strength L of the 3rd color lump and the 4th color lump is measured by detecting light sourceSurvey 3With
LSurvey 4, revised signal strength L is calculated respectivelyRepair 3=kLSurvey 3+ b and LRepair 4=kLSurvey 4+ b, then according to LPhase 3- LRepair 3And LPhase 4- LRepair 4
Absolute value judge whether to need further to correct.
Preferably, the data message for detecting colorimetric card by photodetectors register when system is dispatched from the factory, according to this
Data message establishes colorimetric card colourity and the corresponding optimal response curve of photo detector signal intensity.
Preferably, if LPhase 3- LRepair 3And LPhase 4- LRepair 4In the absolute value of any one be more than specific threshold, carry out further
Photodetector response coefficient amendment.
Preferably, the correction value of photodetector response coefficient calculates as follows:(1) detection light source conduct is used
Light source, photodetector obtain the data message of each gradient region of colorimetric card successively;(2)The amendment of light source is detected, is obtained
To the revised signal strength L of each gradient regionRepair i;(3)By optimal response curve, each gradient region is calculated respectively
Expect signal strength LPhase i;(4)Calculate the correction value ε of the response coefficient of each gradient regioni=LPhase i/LRepair i。
Preferably, for the signal strength L after the light source modification of detection zone measurementRepair, its correct step be:(1) obtain
Its colourity corresponds to the correction value ε of the response coefficient of gradient region endpoint1And ε2;(2)By for ε1And ε2Interpolation calculation is carried out to go out
The correction value ε of the colourityRepair;(3)Calculate signal strength LRepairFinal correct Lf=LRepair ×εRepair。
Preferably, according to light source modification and the revised signal strength of photodetector response coefficient, sample is determined
Target detection thing concentration.
As another aspect of the present invention, there is provided a kind of immunochromatographytest test kit, including the claims
One of described in immunity chromatography detection test paper, and colorimetric card, the colorimetric card mark target detection thing concentration and color gradient
Relation.
Brief description of the drawings
Fig. 1 is the immunity chromatography detection test paper schematic diagram of the embodiment of the present invention.
Embodiment
In order to illustrate more clearly of technical scheme, embodiment will be used simply to be situated between the present invention below
Continue, it should be apparent that, in describing below is only one embodiment of the present of invention, is come for those of ordinary skill in the art
Say, without having to pay creative labor, other technical solutions can also be obtained according to these embodiments, are fallen within
Disclosure of the invention scope.
The immunity chromatography detection test paper of the embodiment of the present invention, referring to Fig. 1, including filling opening 1, detection zone 2 and calibration
Region 3.Wherein, filling opening 1 sets water absorption pad, and for adding detection sample to test paper by filling opening 1, detection sample can be
Body fluids such as blood etc..Detection line is set in detection zone 2, when in sample there are during target detection thing, sample through capillary action,
Nano gold mark composition granule is taken on the p-wire for being adsorbed with antibody, if the antigen in sample is corresponding with this antibody, is produced
Raw specific binding, colloidal gold is trapped on p-wire, in particular color.The light sent by detecting light source is radiated at test strips
Foretell, test strips surface scattering light is received by photodetector.Luminous intensity reflects degree of absorption of the object to incident light, according to bright
Primary-Beer law, luminous intensity and the concentration of target checking matter in sample are linear, so the survey according to photodetector
Amount is as a result, it is possible to determine the concentration of detected material in sample.
Technical scheme is based on following principle, and the aging of detecting system includes aging and the photoelectricity of light-source system
The aging of detector.The aging of wherein electro-optical system is due to that the luminescent properties of light source such as light emitting diode are degenerated, and is caused same
The input parameter of sample, its luminous intensity weaken, and thus cause the error of testing result, its caused error is linear.And light
During electric explorer such as photodiode aging, the change of its photoelectric response performance, causes the response coefficient change for colourity light source
Change, error caused by the change is nonlinear.In the prior art such as 201210185414.7 patents of invention, by using single
Correction factor the aging of light-source system and the aging of photodetector are subjected to simple value revision together, can not react
The physical process of aging, causes correction result undesirable.Also, the digestion period of light source and photodetector be also it is different,
The possible light source ages cycle is shorter and the digestion period of photodetector is longer, it is only necessary to which the aging into line light source is corrected.
The calibration region 3 of immunity chromatography detection test paper in the embodiment of the present invention is used for the aging effect of detecting system, bag
Include the first color lump 31, the second color lump 32, the 3rd color lump 33 and the 4th color lump 34.Wherein first calibration 31 and the second color lump 32 have
There is identical color, for carrying out determining for Light source correction coefficient.First color lump, 31 surface covers specific light transmittance such as 80%
The photic zone of light transmittance, 32 surface of the second color lump do not cover photic zone.
The first color lump 31 and the second color lump 32 are irradiated by detecting light source and standard sources respectively before detection, according to light
Electric explorer in response to determining that detection light source aging amendment.Wherein, the standard sources keeps stabilization for exiting light beam intensity
Light source, it is arranged to can be used in multiple detecting systems far below the correction and replacement cycle, single standard sources that detect light source.
Specifically, the aging amendment of detection light source includes the following steps:1st, basisCorrection factor k and b are calculated, wherein
LMark 1The signal strength that photodetector detects during to irradiate the first color lump 31 using standard sources, LMark 2To be shone using standard sources
The signal strength that photodetector detects when penetrating the second color lump 32;LInspection 1Photoelectricity during to irradiate the first color lump 31 using detection light source
The signal strength of detector detection, LInspection 2The signal that photodetector detects during to irradiate the second color lump 32 using detection light source is strong
Degree;2nd, by correction factor k and b, L is modified to for detecting the signal strength of light source aging effect when detecting sampleRepair=
kLSurvey+ b, wherein L survey the signal strength of photodetector detection during to detect sample.Can be from thing by the above-mentioned light source modification
The degree of aging of light source is reacted during reason completely, realizes the physical process amendment for light source ages, rather than be only numerical value
Fitting.
3rd color lump 33 and the 4th color lump 34 are used for the degree of aging for judging photodetector, determine whether to need
Carry out the aging amendment of photodetector.3rd color lump 33 and the 4th color lump 34 are respectively different colours, wherein the 3rd color lump
33 colors are close to the lower limit of the color range of nano gold mark composition granule, and 34 color of the 4th color lump is close to nano gold mark composition granule
Color range the upper limit.Specifically,(1)It is strong by the signal for detecting light source the 3rd color lump 33 of measure and the 4th color lump 34
Spend LSurvey 3And LSurvey 4, the signal strength L after light source modification is calculated respectivelyRepair 3=kLSurvey 3+ b and LRepair 4=kLSurvey 4+b;(2)Pass through optimal response
Curve, calculates the expectation signal strength of the 3rd color lump 33 and the 4th color lump 34, LPhase 3=k1X1+b1And LPhase 4=k1X2+b1, wherein
K1 and b1 is optimal response slope of a curve and intercept, and X1 and X2 are respectively the color range gradient number of the 3rd color lump and the 4th color lump
Value;(3)The signal strength L of the 3rd color lump 33 and the 4th color lump 34 is measured by detecting light sourceSurvey 3And LSurvey 4, calculate correct respectively
Signal strength L afterwardsRepair 3=kLSurvey 3+ b and LRepair 4=kLSurvey 4+ b, then according to LPhase 3- LRepair 3And LPhase 4- LRepair 4Absolute value judge whether
Need further to correct;(4)If LPhase 3- LRepair 3And LPhase 4- LRepair 4In the absolute value of any one be more than specific threshold, represent
Differing greatly for its response coefficient and expected value, carries out the amendment of further photodetector response coefficient;Otherwise, without
The amendment of photodetector response coefficient.Wherein, optimal response curve negotiating following manner determines:Pass through light when system is dispatched from the factory
The data message of electric explorer record detection colorimetric card, colorimetric card colourity and photo detector signal are established according to this data message
The corresponding optimal response curve of intensity.
If according to the testing result of the 3rd color lump 33 and the 4th color lump 34, determine to need to carry out the old of photodetector
Change and correct, then the correction value of photodetector response coefficient calculates as follows:(1) detection light source is used as light source,
Photodetector obtains the data message of each gradient region of colorimetric card successively;(2)The amendment of light source is detected, is obtained each
The revised signal strength L of gradient regionRepair i;(3)By optimal response curve, the expectation for calculating each gradient region respectively is believed
Number intensity LPhase i;(4)Calculate the correction value ε of the response coefficient of each gradient regioni=LPhase i/LRepair i。
According to the correction value of the response coefficient of each gradient region of acquisition, after the light source modification of detection zone measurement
Signal strength LRepair, its correct step be:(1) the correction value ε that its colourity corresponds to the response coefficient of gradient region endpoint is obtained1With
ε2;(2)By for ε1And ε2Carry out the correction value ε that interpolation calculation goes out the colourityRepair;(3)Calculate signal strength LRepairFinal repair
Positive Lf=LRepair ×εRepair。
After light source modification and the amendment of photodetector response coefficient has been carried out, light source modification and photodetection are obtained
The revised signal strength of device response coefficient, according to the correspondence of signal strength and target detection thing concentration, can determine sample
The concentration of target detection thing in product.
All documents that the present invention refers to all incorporated by reference in this application, work is individually recited just as each document
For with reference to such.In addition, it should also be understood that, after the above disclosure of the present invention has been read, protection scope of the present invention and not only
Above-described embodiment is confined to, those skilled in the art can make various modifications or changes to the present invention, is not departing from original of the invention
Under the premise of reason, these equivalent forms also fall within the scope of the appended claims of the present application.