CN107525926A - A kind of immunochromatography detection method and detecting system - Google Patents
A kind of immunochromatography detection method and detecting system Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
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Abstract
A kind of detection method and detecting system of immunochromatography detecting system, comprise the following steps：（1）The calibration color lump irradiated respectively by standard sources and detection light source in calibration region, according to photodetector in response to determining that the correction factor of detection light source；（2）Determine the need for carrying out the amendment of photodetector response coefficient according to the result of the amendment of detection light source；Enter step if necessary to correct（3）If it need not correct into step（4）；（3）It is modified for photodetector response coefficient；（4）By detecting light source irradiating and detecting region, revised signal intensity is obtained；（5）According to revised signal intensity, the target detection thing concentration of sample is determined.
Description
Technical field
The present invention relates to immuno-chromatographic assay technology field, more particularly, to immunochromatography detecting system.
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.Collaurum 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.Because collaurum is made
Had 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 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 chromatographs detecting system 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, collaurum 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, collaurum is trapped on p-wire, in particular color.Utilize entering to Test paper with light source and photodetector
The instant interpretation of row, can realize and target checking matter is quantitatively detected.
Conventional technique scheme, irradiation light is formed 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 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 establishes the optimal response curve corresponding with 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, pass through the correction factor amendment system
The result for components aging of uniting.But also there is following defect in the technical scheme 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, its amendment is only value revision,
Specific physical process is not considered, the result is that nonlinear, accuracy deficiency.
The content of the invention
The invention provides improved technical scheme, can solve the problem that the above mentioned problem of prior art, and improves amendment
Accuracy.
As one aspect of the present invention, there is provided a kind of immunochromatography detecting system, including：Light source is detected, photoelectricity is visited
Survey device, immunity chromatography detection test paper；The immunity chromatography detection test paper sets detection zone, and the detection zone sets detection
Line, when target detection thing in sample be present, nano gold mark composition granule therein occurs immunochromatography reaction and is gathered in detection
On line；The detection light source can launch detection light to the detection zone；The photodetector can receive the detection
The reflected light in region；It is characterized in that：Also include standard sources, it can be to the detection zone emission standard light；It is described to exempt from
Epidemic disease chromatography detecting test paper sets calibration region, and the calibration region sets calibration color lump, and the calibration color lump is located at the nanometer
The color range of gold mark composition granule, the detection irradiated by standard light and detection light for calibration color lump are entered for testing result
Row is corrected and determines the need for further correcting.
Preferably, the standard sources is that can provide the light source of stable emergent light intensity, and it is arranged to far below detection
The correction of light source and replacement cycle.
Preferably, in addition to colorimetric card, it identifies target detection thing concentration and the relation of color gradient.
Preferably, memory module, it stores the optimal response corresponding with photo detector signal intensity of colorimetric card colourity
Curve.
Preferably, the calibration color lump includes the color lump of color identical first and the second color lump, 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, the calibration color lump also includes the 3rd color lump and the 4th color lump, the 3rd color lump and the 4th color
Block is respectively different colours, wherein lower limit of the 3rd color lump color close to the color range of the nano gold mark composition granule, the
The upper limit of the four color lump colors close to the color range of the nano gold mark composition granule.
As another aspect of the present invention, there is provided the detection method of immunochromatography detecting system, comprise the following steps：
（1）The calibration color lump irradiated respectively by standard sources and detection light source in calibration region, it is true according to the response of photodetector
The correction factor in regular inspection light-metering source；（2）Determine the need for carrying out photodetector sound according to the result of the amendment of detection light source
Answer the amendment of coefficient；Enter step if necessary to correct（3）If it need not correct into step（4）；（3）Visited for photoelectricity
Device response coefficient is surveyed to be modified；（4）By detecting light source irradiating and detecting region, revised signal intensity is obtained；（5）According to
Revised signal intensity, determine the target detection thing concentration of sample.
Preferably, the step（1）In, irradiate first color lump and the by detecting light source and standard sources respectively
Two color lumps, according to photodetector in response to determining that the amendment of detection light source；The amendment comprises the following steps：1st, basisCalculate correction factor k and b, wherein LMark 1Photodetector is examined during to irradiate the first color lump using standard sources
The signal intensity of survey, LMark 2The signal intensity that photodetector detects during to irradiate the second color lump using standard sources；LInspection 1To use
Detect the signal intensity of photodetector detection when light source irradiates the first color lump, LInspection 2To irradiate the second color lump using detection light source
When photodetector detection signal intensity；2nd, when detecting light source irradiating and detecting region, photodetection corresponding to light source is detected
The signal intensity of device detection is modified to LRepair=kLSurvey+b。
Preferably, the step（2）In, by optimal response curve, calculate the expectation of the 3rd color lump and the 4th color lump
Signal intensity, LPhase 3=k1X1+b1And LPhase 4=k1X2+b1, wherein k1 and b1 are optimal response slope of a curve and intercept, X1 and X2
The respectively color range gradient magnitude of the 3rd color lump and the 4th color lump；The 3rd color lump and the 4th color lump are determined by detecting light source
Signal intensity LSurvey 3And LSurvey 4, revised signal intensity L is calculated respectivelyRepair 3=kLSurvey 3+ b and LRepair 4=kLSurvey 4+ b, if LPhase 3- LRepair 3With
LPhase 4- LRepair 4In the absolute value of any one be more than specific threshold, carry out the amendment of further photodetector response coefficient.
Preferably, the step（3）The correction value of middle photodetector response coefficient calculates as follows:(a) make
The data message of each gradient region of colorimetric card is obtained successively as light source, photodetector by the use of detection light source；（b）Detected
The amendment of light source, obtain the revised signal intensity L of each gradient regionRepair i；（c）By optimal response curve, calculate respectively each
The expectation signal intensity L of individual gradient regionPhase i；（d）Calculate the correction value ε of the response coefficient of each gradient regioni=LPhase i/LRepair i。
Preferably, in the step (4), the signal intensity for detecting photodetector detection corresponding to light source is modified to LRepair=
kLSurvey+b；According to step（2）Result judgement whether need the amendment of photodetector response coefficient, it is if necessary to correct, then right
In LRepairThe amendment step of photodetector response coefficient be：（a1）Obtain the response coefficient that its colourity corresponds to gradient region end points
Correction value ε1And ε2；（a2）By for ε1And ε2Carry out the correction value ε that interpolation calculation goes out the colourityRepair；（a3）Calculate the signal
Intensity LRepairFinal amendment Lf=LRepair ×εRepair。
Brief description of the drawings
Fig. 1 is the immunochromatography detecting system schematic diagram of the embodiment of the present invention.
Fig. 2 is the overhaul flow chart of the immunochromatography detecting system 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 schemes can also be obtained according to these embodiments, fallen within
Disclosure of the invention scope.
The immunochromatography detecting system of the embodiment of the present invention, referring to Fig. 1, including immunity chromatography detection test paper 10, detection light
Source 20, standard sources 30 and photodetector 40.
Detect light source 20 to be used to provide incident light to immunity chromatography detection test paper 10, received by using photodetector 40
The scattering light of immunity chromatography detection test paper 10, such as LED light source can be used as detection light source.Standard sources 30 is emergent light
Intensity keeps stable light source, and it is arranged to correction and replacement cycle far below detection light source 20, the single energy of standard sources 30
It is enough in multiple detecting systems.
Immunity chromatography detection test paper 10 includes filling opening 1, detection zone 2 and calibration region 3.Filling opening 1 sets water suction
Pad, for adding detection sample to test paper by filling opening 1, detection sample can be body fluids such as blood etc..Set in detection zone 2
Detection line is put, when target detection thing in sample be present, sample takes nano gold mark composition granule to absorption by capillarity
Have on the p-wire of antibody, if the antigen in sample is corresponding with this antibody, produce specific binding, collaurum is trapped in survey
Try on line, in particular color.The light irradiation sent by detecting light source is foretold in test strips, and test strips surface scattering light is visited by photoelectricity
Device is surveyed to receive.Luminous intensity reflects degree of absorption of the object to incident light, according to Lambert-Beer's law, luminous intensity and mesh in sample
It is linear to mark the concentration of checking matter, so according to the measurement result of photodetector, can determine detected material in sample
Concentration.
The calibration region 3 of immunity chromatography detection test paper is used for the aging effect of detecting system, including the first color lump 31, second
Color lump the 32, the 3rd calibrates color lump 33 and the 4th and calibrates color lump 34.Wherein the first color lump 31 and the second color lump 32 have identical
Color, for carrying out the determination of Light source correction coefficient.The surface of first color lump 31 covers specific light transmittance such as 80% light transmittance
Photic zone, the surface of the second color lump 32 do not cover photic zone.
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 the color of the 4th color lump 34 is close to nano gold mark composition granule
Color range the upper limit.
Immunochromatography detecting system also includes colorimetric card and memory module, and wherein colorimetric card is dense for identifying target detection thing
Degree and the relation of color gradient, memory module are used to storing optimal corresponding with photo detector signal intensity of colorimetric card colourity
Response curve.Wherein, optimal response curve negotiating following manner determines：Detected when system is dispatched from the factory by photodetectors register
The data message of colorimetric card, optimal corresponding with photo detector signal intensity of colorimetric card colourity is established according to this data message
Response curve.
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 light-source 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, cause the response coefficient change for colourity light source
Change, error caused by the change is nonlinear.In 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, cause 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 enters the aging amendment of line light source.
Referring to Fig. 2, the detection method of the immunochromatography detecting system of the embodiment of the present invention, comprise the following steps：（1）Respectively
The calibration color lump irradiated by standard sources 30 and detection light source 20 in calibration region 3, it is true according to the response of photodetector 40
The correction factor in regular inspection light-metering source 20；（2）Determine the need for carrying out photodetection according to the result of the amendment of detection light source 20
The amendment of the response coefficient of device 40；Enter step if necessary to correct（3）If it need not correct into step（4）；（3）For
The response coefficient of photodetector 40 is modified；（4）By detecting the irradiating and detecting region 2 of light source 20, revised signal is obtained
Intensity；（5）According to revised signal intensity, the target detection thing concentration of sample is determined.
The step（1）In, irradiate the first color lump 31 and the second color by detecting light source 20 and standard sources 30 respectively
Block 32, according to photodetector 40 in response to determining that the amendment of detection light source 20；The amendment comprises the following steps：1st, basisCalculate correction factor k and b, wherein LMark 1Photodetection during to irradiate the first color lump 31 using standard sources 30
The signal intensity that device 40 detects, LMark 2The signal that photodetector 40 detects during to irradiate the second color lump 32 using standard sources 30
Intensity；LInspection 1The signal intensity that photodetector 40 detects during to irradiate the first color lump 31 using detection light source 20, LInspection 2To use inspection
The signal intensity that photodetector 40 detects when the second color lump 32 is irradiated in light-metering source 20；2nd, in detection light source 20 irradiating and detecting region
When 2, detect the signal intensity that photodetector 40 corresponding to light source 20 detects and be modified to LRepair=kLSurvey+b。
The step（2）In, by optimal response curve, the expectation signal of the 3rd color lump 33 of calculating and the 4th color lump 34
Intensity, LPhase 3=k1X1+b1And LPhase 4=k1X2+b1, wherein k1 and b1 are optimal response slope of a curve and intercept, and X1 and X2 distinguish
For the 3rd color lump 33 and the color range gradient magnitude of the 4th color lump 34；The 3rd color lump 33 and the 4th color are determined by detecting light source 20
The signal intensity L of block 34Survey 3And LSurvey 4, revised signal intensity L is calculated respectivelyRepair 3=kLSurvey 3+ b and LRepair 4=kLSurvey 4+ b, if
LPhase 3- LRepair 3And LPhase 4- LRepair 4In the absolute value of any one be more than specific threshold, carry out further photodetector 40 and respond
The amendment of coefficient.
The step（3）The correction value of the middle response coefficient of photodetector 40 calculates as follows:(a) using detection
Light source 20 is used as light source, and photodetector 40 obtains the data message of each gradient region of colorimetric card successively；（b）Carry out detection light
The amendment in source 20, obtain the revised signal intensity L of each gradient regionRepair i；（c）By optimal response curve, calculate respectively each
The expectation signal intensity L of individual gradient regionPhase i；（d）Calculate the correction value ε of the response coefficient of each gradient regioni=LPhase i/LRepair i。
In the step (4), detect the signal intensity that photodetector 40 corresponding to light source 20 detects and be modified to LRepair=kLSurvey
+b；According to step（2）Result judgement whether need the amendment of the response coefficient of photodetector 40, it is if necessary to correct, then right
In LRepairThe amendment step of the response coefficient of photodetector 40 be：（a1）Obtain the response system that its colourity corresponds to gradient region end points
Several correction value ε1And ε2；（a2）By for ε1And ε2Carry out the correction value ε that interpolation calculation goes out the colourityRepair；（a3）Calculate the letter
Number intensity LRepairFinal amendment Lf=LRepair ×εRepair。
After light source modification and photodetector response coefficient amendment has been carried out, light source modification and photodetection are obtained
The revised signal intensity of device response coefficient, according to the corresponding relation of signal intensity and target detection thing concentration, it 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 is to 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 changes or modifications to the present invention, not depart from original of the invention
Under the premise of reason, these equivalent form of values equally fall within the application appended claims limited range.
Claims (3)
1. a kind of detection method of immunochromatography detecting system, comprises the following steps：（1）Pass through standard sources and detection light respectively
Calibration color lump in source irradiation calibration region, according to photodetector in response to determining that the correction factor of detection light source；（2）According to
The result for detecting the amendment of light source determines the need for carrying out the amendment of photodetector response coefficient；If necessary to correct entrance
Step（3）If it need not correct into step（4）；（3）It is modified for photodetector response coefficient；（4）Pass through inspection
Light-metering source irradiating and detecting region, obtains revised signal intensity；（5）According to revised signal intensity, the mesh of sample is determined
Mark detectable substance concentration；The step（2）In, by optimal response curve, the expectation for calculating the 3rd color lump and the 4th color lump is believed
Number intensity, LPhase 3=k1X1+b1And LPhase 4=k1X2+b1, wherein k1 and b1 are optimal response slope of a curve and intercept, and X1 and X2 divide
Not Wei the 3rd color lump and the 4th color lump color range gradient magnitude；The letter of the 3rd color lump and the 4th color lump is determined by detecting light source
Number intensity LSurvey 3And LSurvey 4, revised signal intensity L is calculated respectivelyRepair 3=kLSurvey 3+ b and LRepair 4=kLSurvey 4+ b, if LPhase 3- LRepair 3With
LPhase 4- LRepair 4In the absolute value of any one be more than specific threshold, carry out the amendment of further photodetector response coefficient.
2. the detection method of immunochromatography system according to claim 1, it is characterised in that：The step（3）Middle photoelectricity
The correction value of explorer response coefficient calculates as follows:(a) using detection light source, photodetector is successively as light source
Obtain the data message of each gradient region of colorimetric card；（b）Detect the amendment of light source, after obtaining each gradient region amendment
Signal intensity LRepair i；（c）By optimal response curve, the expectation signal intensity L of each gradient region is calculated respectivelyPhase i；（d）Meter
Calculate the correction value ε of the response coefficient of each gradient regioni=LPhase i/LRepair i。
3. a kind of immunochromatography detecting system, it is used for the detection method of one of the claims.
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| CN110579598B (en) * | 2018-06-07 | 2023-12-05 | 昆明联恩生物技术有限责任公司 | Optical signal detection device, system and method |
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|---|---|
| CN107525926B (en) | 2020-03-27 |
| CN105974108B (en) | 2018-05-01 |
| CN105974108A (en) | 2016-09-28 |
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