CN201535749U - Quantum dot mark test strip quantitative detection system based on CMOS picture sensor - Google Patents
Quantum dot mark test strip quantitative detection system based on CMOS picture sensor Download PDFInfo
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- CN201535749U CN201535749U CN2009200073261U CN200920007326U CN201535749U CN 201535749 U CN201535749 U CN 201535749U CN 2009200073261 U CN2009200073261 U CN 2009200073261U CN 200920007326 U CN200920007326 U CN 200920007326U CN 201535749 U CN201535749 U CN 201535749U
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Abstract
The utility model relates to a quantum dot mark test strip quantitative detection system based on a CMOS picture sensor, which comprises a quantum dot mark test strip, a test strip rack, a scanning system, the CMOS picture sensor, a data processing device, an output display device, a printer, a keyboard, and an IC card matched with the test strip, wherein the data processing device reads parameters of the IC card and then controls the movement of the test strip rack, so as to adapt to the automatic scanning of a fiber-optics probe of the scanning system to a detection band and a quality control band; the collected characteristic wavelength reflects a fluorescence signal; the fluorescence signal is transmitted to the data processing device via the CMOS picture sensor to recognize the optical density and calculate the concentration; and the output display device displays the detected results. The utility model can quickly and accurately realize the quantitative or qualitative detection of single components and multicomponents of samples, and can also dynamically monitor the immuno-chromatographic reaction process. The system has the characteristics of high detection sensitivity, objectivity of results, flexible use and the like.
Description
Technical field
The utility model belongs to the biologic medical instrument field, is specifically related to a kind of quantitative detection system of quantum dot mark test strip and method thereof based on cmos image sensor.This system can scan quanta dot mark test bar and detect and interpretation as a result, realizes the quantitative or qualitative detection to plurality of target checking matters such as pathogen, antigen, antibody, illegal drug, major disease (tumour, cancer, angiocardiopathy and diabetes etc.), persticide residue, food security biological detection.
Background technology
In immune detection, the main at present methods such as radioimmunoassay method, enzyme linked immunosorbent assay (ELISA) and colloid gold label that adopt realize the quantitative or qualitative detection of sample.There is defective in various degree in these methods.
Quantum dot (Quantum Dot, be called for short QD) be a kind of semiconductor nanocrystal of the development nineties in 20th century with good spectral signature and photochemical stability, have luminescence efficiency height, exciting line wide ranges, spectral line of emission narrow range, particle diameter and characteristics such as biomolecule is close, finishing multifunction.As biological fluorescent labelling thing of new generation, the interaction between the simultaneous determination of multiponents of quantum dot in the labelled immune analysis, immune tracer molecule location, nano biological sensor, detection in real time, cell imaging, in-vivo imaging, medical diagnosis on disease and the research biomacromolecule, component circulation and the aspect such as the mode of action in body has unique effect and wide application prospect.Bibliographical information, quantum dot have the far-reaching potentiality that replace traditional organic dyestuff as biological fluorescent labelling thing of new generation.Compare with organic dye molecule, the major advantage of QD has:
(1) good light stability.Under excitation light irradiation, the often very fast deepening of the fluorescence signal of organic dyestuff, quantum dot then can continue luminous, and photobleaching speed is 1/100 of rhodamine.
(2) luminous intensity height.Quantum dot is the polyelectron system, and luminescence efficiency is far above single organic molecule, and it is 10 in the absorptivity of visible and ultraviolet region
5L/ (mol.cm) order of magnitude makes its fluorescent emission intensity far above organic dyestuff.
(3) excitation spectrum wide ranges.Common organic fluorescence molecule usually has specific excitation wavelength and emission wavelength, must be with the light deexcitation of several wavelength when measuring several fluorescence spectrum, and this analyzes for multiple labeling simultaneously and has caused great trouble.And the excitation spectrum of quantum dot almost is continuous absorbing more than the threshold values, and spectrum peak broad only needs a light source activation than any wavelength of its wavelength of transmitted light weak point, can launch the fluorescence of high brightness, and can be detected simultaneously.
(4) the bigger Stokes displacement and the luminescent spectrum of narrow symmetry.The narrow symmetry of the relative organic dyestuff peak shape of the fluorescence spectra of quantum dot, the high width of half-peak has only 1/3 of fluorescein, peak width at half height usually has only 40nm or littler, and there is not conditions of streaking in the long wave direction, this allows to use simultaneously the quantum dot of different spectral signatures, overlapping does not appear in emission spectrum, makes the differentiation of mark biomolecule fluorescence Spectra, identification become and be easy to, and this detects simultaneously to biomedical sector realization sample and brings great convenience.
(5) fluorescence lifetime is longer, is about hundreds of nanoseconds.Fluorescent bleach speed has only 1/100 of rhodamine 6G, and higher photochemical stability can guarantee that it retains a couple of days and even several months in vivo.Bibliographical information is arranged, in the mouse body, inject quantum dot, after 8 weeks, still have fluorescence to detect.
Quantum dot has extremely wide application prospect as biological fluorescent labeling in immune detection.The quantum dot of different material and the different particle sizes of same raw material can produce the emitting fluorescence of different wave length, and the quantum dot potpourri does not produce variable spectrum fluorescence.This quantum dot optics characteristic that produces specific fluorescent according to the quantum dot of specific size, composition and structure is come that quantum dot-labeled immunity-chromatography test strip is carried out specific fluorescent and is measured, and can accurately detect the concentration of polycomponent checking matter.
Chinese patent application numbers 200610024086.7 has disclosed a kind of quantum dot-labeled fast immune chromatographic strip detection method, the antibody of the quantum dot-labeled different target thing of this application patent utilization different-grain diameter or kind, the testing result that can detect by an unaided eye under ultraviolet realizes that the sample polycomponent detects simultaneously.Weak point is: (1) this application patent only is described the making of quanta dot mark test bar, and do not disclose corresponding supporting detection system, its detection need just can be carried out by other Ultraviolet Detector, and is qualitative detection, can not realize detection by quantitative.(2) adopt ultraviolet detection harmful, operate dangerous.(3) the measurement result objectivity is poor.Because the tester observes the strong and weak degree of fluorescence that the strip detection is with and quality control band produces by naked eyes to judge testing result under ultraviolet, testing result is subject to the influence of tester's experience, subjectivity is strong, same strip might be made different or even wrong judged result under different conditions, particularly for the extremely low sample of those object content, the very weak judged result with the naked eye that just is difficult to of the fluorescence that ultraviolet is sent, sensitivity is low.(4) can not monitoring bio the dynamic process of reaction.Immunochromatography is a dynamic reaction process, being in siphon on the strip oozes the quantum dot-labeled particle of shifting state and produces fluorescence and belong to dynamic category, the ultraviolet visual method obviously can not be observed and analyze this dynamic reaction process, so can't more in depth study the immunochromatography course of reaction.Even biological respinse failure, the tester is the reason of analytical reactions failure well also.
Chinese patent application numbers 200610024566.3 has disclosed a kind of quantum dot fluorescence probe biochip for joint detection and has sought Chinese medicine target spot method, there is deficiency equally in this patent: the detection of " quantum dot-traditional Chinese medicine ingredients " fluorescence probe that it relates to not only needs other technology that the corresponding protein chip is provided, and its detection equally need be by other specialized instrument and equipment.
Chinese patent application numbers 200510059892.3 has disclosed and has been used for the optoelectronic rapid diagnostic test system that the quantum dot strip detects a kind of comprising, go out off-the-shelf but this patent is still untapped at present, and content is different fully with the utility model.
The utility model content
Technical solutions of the utility model are as follows:
The IC-card 19 that quantitative detection system of quantum dot mark test strip based on cmos image sensor described in the utility model comprises quanta dot mark test bar 1, is used to place the strip frame 2 of strip, scanning system 3, cmos image sensor 13, data processing equipment 15, output display device 16, printer 17, keyboard 18 and one and strip 1 are supporting.
The strip frame 2 of described system has the strip groove.The strip groove is used to place quanta dot mark test bar to be measured 1.Described strip 1 is provided with sample pad 20, the glass fibre membrane pad 21 that is coated with quantum dot-labeled thing that mutual overlap joint pastes in turn, have and detect with 26 and the analyzing film 22 of quality control band 27, strong adsorptive pads 23, strip reaction end indicating label 24 and strip identification label 25.
Described scanning system 3 comprises an excitation source 4, is followed successively by incident light coupling mechanism 5, incident optical 6,7, fibre-optical probe 8,9, outgoing optical fiber 10,11, emergent light coupling mechanism 12 on the output light path of this excitation source 4, until cmos image sensor 13.Described light source 4 comprises LED or laser diode.Described fibre-optical probe 8,9 closes Shu Gongyong, and its center is an incident optical 28, and periphery is an outgoing optical fiber 29.
The signal output part of described cmos image sensor 13 links to each other with the signal input part of data processing equipment 15, cmos image sensor 13 be used for detection with 26 and the quality control band 27 transmission quantum dot-labeled compound characteristic wavelength reflected fluorescence signal of coming carry out photoelectricity and mould/number conversion, thereby become digital signal.
Described data processing equipment 15 is gathered, the next digital signal of storage cmos image sensor 13 transmission, and digital signal is carried out data processing.Described data processing equipment 15 also is furnished with output display device 16, printer 17, keyboard 18 and an IC-card that matches with strip 19, and they link to each other with data processing equipment 15 respectively.Also has a Drive and Control Circuit 30 between described data processing equipment 15 and the strip frame 2,31, described data processing equipment 15 reads IC-card 19 parameters and sends instruction by this Drive and Control Circuit 30,31 automatically 2 motions of control strip framves with 8, the 9 pairs of strips of fibre-optical probe that are fit to scanning system 3 detect be with 26 and quality control band 27 carry out autoscan.Described data processing equipment 15 can corresponding data be handled and single-chip microcomputer, CPU or the PC of Control Software in order to have.
The output display device 16 of described detection by quantitative system can be host computer, alphanumeric LCD, LED, sound etc.Described detection by quantitative system cooperates different output display devices 16, realizes that sample single component or polycomponent are quantitatively or qualitative detection: when described detection by quantitative system was used for the sample detection by quantitative, output display device 16 can be host computer or alphanumeric LCD; When described detection by quantitative system was used for the sample qualitative detection, output display device 16 can be LED or sound etc.
Adopt the USB serial communication between described IC-card 19 and the data processing equipment 15.During sample detection, IC-card 19 can plug in real time.IC-card 19 is stored each checking matter respective detection standard working curve and strip quality control band optical density reference monitor value.Each checking matter respective detection standard working curve that IC-card 19 is stored can adopt one of following two kinds of curve modes: the ratio (OD of one detection band optical density value/quality control band optical density value that to be checking matter standard items series concentration record through detection by quantitative described in the utility model system with the serial concentration of this checking matter standard items
Detect band/ OD
Quality control band) between the corresponding relation curve; Also ratio (the OD of detection band optical density value/(detect and be with optical density value+quality control band optical density value) that can record through detection by quantitative described in the utility model system for checking matter standard items series concentration and this checking matter standard items series concentration
Detect band/ (OD
Detect band+ OD
Quality control band)) between the corresponding relation curve.The strip quality control band 26 optical density value (OD that obtain during standard curve making
Quality control band) be stored in simultaneously the reference monitor value of double as strip reaction success or failure on the IC-card 19 with standard working curve.In the sample actual detected, if the reaction failure, the OD that sample detection obtains
Quality control bandThe corresponding OD that stores on value and the IC-card 19
Quality control bandValue will produce very big statistical errors.IC-card 19 is stored parameter and is matched with each batch strip product.During sample detection, the operator inserts IC-card 19, data processing equipment 15 sends the fibre-optical probe 8 that 2 motions of the automatic control of instruction strip frame make scanning system 3 after reading IC-card 19 parameters, the 9 pairs of strips 1 detect be with 26 and quality control band 27 carry out autoscan, thereby obtain detecting with 26 and the characteristic frequency fluorescence optical density value sent of quality control band 27, and can realize that according to the corresponding checking matter standard working curve (simultaneously with reference to the suggested biological respinse success or failure of strip quality control band optical density reference monitor value) of IC-card 19 storages checking matter concentration accurately measures.IC-card 19 also can be made temporary storage storage testing result information, and each detection can be printed or preserve.
The method for making of each checking matter respective standard working curve of described IC-card 19 storages is: (1) preparation standard items series concentration; (2) each standard items series concentration is placed detection by quantitative described in the utility model system record corresponding OD
Detect bandAnd OD
Quality control bandAnd calculate OD respectively
Detect band/ OD
Quality control bandRatio or OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio; (3) make X-axis with each standard items series concentration, test result is with OD
Detect band/ OD
Quality control bandRatio is made Y-axis, the drawing standard working curve; Or: make X-axis with each standard items series concentration, test result is with OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio makes Y-axis, the drawing standard working curve; (4) write standard working curve software, and this Curve Software and strip quality control band optical density reference monitor value are stored on the IC-card 19.
One synchronous control circuit (showing in the Figure of description) can also be arranged to avoid 13 integral time of cmos image sensor long and cause noise level to improve between the light source 4 of described detection by quantitative system and the cmos image sensor 13.
Can send characteristic wavelength fluorescence after the optical excitation that the principle that detects the utility model is transmitted by incident optical 6,7 with the quantum dot-labeled compound on 26 based on: the detection of quanta dot mark test bar, this fluorescence signal becomes positive correlation with testing sample concentration.If testing sample concentration is big more in the sample, it is just strong more that strip detects the characteristic wavelength fluorescence signal that sends with the quantum dot-labeled compound on 26.Otherwise testing sample concentration is more little in the sample, and it is just weak more that strip detects the characteristic wavelength fluorescence signal that sends with the quantum dot-labeled compound on 26.According to this relation, the utility model is applied to testing sample in the strip one end well 14, corresponding strip sample pad 20 under the well 14, utilize the capillary siphoning effect of strip miillpore filter, target checking matter fluid sample is slowly oozed to strip miillpore filter rear end under the pulling of the strong adsorptive pads 23 of the strip other end to be moved, when target checking matter molecule (as checking matter specific antigen or antibody) is arranged in the fluid sample, they just the corresponding biomolecule that is labeled quantum dot on the glass fibre membrane pad 21 that is coated on the preceding stage casing of strip in conjunction with and ooze the detection that moves to strip analyzing film 22 to the strip rear end and be with 26, be with quantum dot-labeled compound, the quantum dot-labeled thing of a remaining part to continue to ooze to move to strip quality control band 27 and to combine the formation quality control band quantum dot-labeled multiple with the Quality Control thing (anti-as two) of the pre-bag quilt of quality control band 27 positions with being coated on to detect combine to form to detect with 26 special target checking matter correlation molecule (as the specific antibody or the antigen of respective objects checking matter molecule) ++-++ compound.Be combined in strip detect with 26 and the quantum dot-labeled compound of quality control band 27 because of being combined with quantum dot, they are at incident optical 6, under the exciting of 7 light that transmit, promptly launch corresponding long-life characteristic wavelength fluorescence, this characteristic wavelength fluorescence intensity can reflect checking matter concentration.By receiving, change, transmit, handle this characteristic wavelength fluorescence intensity information, therefore can accurately detect checking matter concentration.
The preparation method of quanta dot mark test bar described in the utility model may further comprise the steps:
(1) each assembly preparation of strip
1. the sample pad preparation selects for use cellulose membrane as the sample pad solid phase material, be cut into film piece with certain specification, sample pad film piece is put into sample pad confining liquid (pH=7.20.03mol/L phosphate buffer+5%BSA, + 0.1%Tween 20) the middle immersion, take out, oven dry makes its abundant drying for standby.
2. the preparation of glass fibre membrane pad selects for use glass fibre membrane as the pad solid phase material, be cut into film piece with certain specification, on this film piece, add the bond solution (being quantum dot-labeled thing solution) of quantum dot and target checking matter correlation molecule, the baked film piece makes its abundant drying for standby.
3. the analyzing film preparation selects for use nitrocellulose filter as the analyzing film solid phase material, be cut into film piece with certain specification, the relevant special molecular of the checking matter of specking target respectively separated by a distance is as detecting band and specking two anti-Quality Control things as quality control band on this film piece, dry this film piece, make its abundant drying for standby.
4. strong adsorptive pads preparation is selected cellulose membrane for use and is cut into the film piece with certain specification, fully drying for standby.
5. to select color change interval for use be the accurate pH test paper of 5.0-9.0 for strip reaction end indicating label preparation, is cut into the film piece with certain specification, fully drying for standby.
6. strip is debated and is known the label preparation and select for use the label paper that is printed with the test paper sign to be cut into the film piece with certain specification, fully drying for standby.
(2) strip assembling each component of strip that will prepare by sample pad 20, glass fibre membrane pad 21, analyzing film 22, strong adsorptive pads 23, strip reaction end indicating label 24, strip debate know label 25 in turn mutually overlap joint stick on the plastic back plate, and cut into the strip of certain specification.This strip can be packed in the shell, and is standby under drying condition.This shell can be plastics.
The quantum dot of quanta dot mark test bar 1 described in the utility model is a semiconductor nano material, comprising: ZnS, CdS, HgS, ZnSe, CdSe, HgSe, CdTe, ZnTe, ZnO, PbSe, HgTe, CaAs, InP, InAs, InCaAs, CdS/ZnS, CdS/Ag
2S, CdS/PbS, CdS/Cd (OH)
2CdS/HgS, CdS/HgS/CdS, ZnS/CdS, ZnS/CdS/ZnS, ZnS/HgS/ZnS/CdS, CdSe/CdS, CdSe/ZnS, CdSe/ZnSe, CdSe/CuSe, CdSe/HgTe, CdSe/HgSe, CdSe/HgSe/CdSe, CdTe/HgS, CdTe/HgTe, InAs/InP, InAs/CdSe, InAs/ZnSe, MgS, MgSe, MgTe, CaS, CaSe, CaTe, SrS, SrSe, SeTe, BaS, BaSe, BaTe, CdS:Mn, ZnS:Mn, CdS:Cu, ZnS:Cu, CdS:Tb, the combination of any one among the ZnS:Tb or any several nano particles, and be nuclear by above-mentioned any one quantum dot, silicon dioxide is the core-shell type nano-complex particle of shell.Above-mentioned quantum dot-labeled target checking matter comprises plurality of target checking matters such as pathogen, antigen, antibody, illegal drug, major disease (tumour, cancer, angiocardiopathy and diabetes etc.), persticide residue, food security biological detection.The utility model carries out single to above-mentioned quantum dot-labeled target checking matter or mixes quantitative or qualitative detection.
The course of work of detection by quantitative described in the utility model system is: the quanta dot mark test bar to be measured 1 that will be added with sample is put in the strip groove of strip frame 2; Data processing equipment 15 send instruction automatically 2 motions of control strip frame make the detection of 8,9 pairs of strips of fibre-optical probe of scanning system 3 be with 26 and quality control band 27 carry out autoscan; The light that light source sends enters incident optical 6 through incident light coupling mechanism 5,7, incident optical 6, bundle was through fibre-optical probe 8 in 7 minutes two, 9 shine respectively strip detect with 26 and quality control band 27 on the quantum dot-labeled compound of excitation-detection band 26 and quality control band 27 send the characteristic wavelength reflected fluorescent light, two bundle reflected fluorescent light are with corresponding outgoing optical fiber 10 with quality control band respectively after testing, 11, emergent light coupling mechanism 12 backs are received by cmos image sensor 13, after cmos image sensor 13 carries out photoelectricity and mould/number conversion with light signal, store and data processing for data processing equipment 15 digital data transmission after the conversion, obtain each checking matter detection and be with 26 optical density value (OD
Detect band) and quality control band 27 optical density value (OD
Quality control band), and then calculate OD
Detect band/ OD
Quality control bandRatio or OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio, and calculate checking matter concentration according to the corresponding checking matter standard working curve analysis meter of storage on the IC-card 19 automatically, testing result is transferred to output display device 16, finishes whole testing process.
Immunochromatography is a dynamic reaction process, and the siphon on strip of quantum dot-labeled particle is oozed and moved the process that produces reflected fluorescent light thereby also belong to dynamic category.The strip product of same batch of different branches of the strip product that different manufacturers is produced or the different batches strip product of same manufacturer production even same manufacturer production, because manufacture craft and raw material selection aspect may there are differences, the quality of the strip product that it is produced is may not can identical.When these strip products were used for the checking matter concentration determination, this will certainly have influence on the accurate mensuration of checking matter concentration.For example, make used cellulose membrane source difference of strip, membrane filtration hole size difference, strip thickness and reactant can not oozed with identical siphon move speed and move ahead on the strip of made, the reactant that has in addition might be trapped in strip detect with 26 and quality control band 27 before strip ooze and move on the way.In addition, environmental baseline of living in can not be just the same during tester's test sample, for example, differences such as the outdoor on-the-spot temperature that detects and exist during indoor detection, humidity also might have influence on the strip reactant and can not move speed and move ahead with identical oozing on strip, thereby have influence on the accurate mensuration of checking matter concentration.For addressing the above problem, the utility model adopts strip with joining the IC-card 19 corresponding checking matter standard working curves of storage and providing strip quality control band optical density reference monitor value prompting reaction success or failure way by IC-card 19.Typical curve on the IC-card 19 and strip quality control band optical density reference monitor value can adjust with the strip differences between batches, operates not only very easyly, and improved accuracy and the dirigibility that detects.Detection can obtain accurate testing result by looking into standard working curve on the IC-card 19; Simultaneously, in conjunction with the strip quality control band optical density reference monitor value of storage on the IC-card 19, can know whether the strip course of reaction is set up, thereby whether judge detected result authentic and valid.In addition, the utility model is also found through a large amount of strip reaction tests: strip added example reaction after 3 minutes, strip reaction end indicating label 24 on the strip can develop the color, the prompting strip is adding a certain particular moment of example reaction after 3 minutes, and the reactant on the strip fully oozes to move past to detect on strip is with 26 and quality control band 27 positions.Therefore, the utility model adopts in 3 minutes later particular moments of described detection by quantitative system boot and just formally makes described system begin sample detection, this relatively more suitable described system IC-card 19 standard working curves and the work of strip quality control band optical density reference monitor value.The utility model fits to System self-test sweep time with this strip reaction time, and this time parameter also can be identified on the IC-card 19 that matches with strip.
The detection method of detection by quantitative described in the utility model system, realize by following step:
1. the energized of starting shooting;
2. insert the IC-card 19 that storage has checking matter examination criteria working curve and strip quality control band optical density reference monitor value;
3.) quanta dot mark test bar 1 that will be added with testing sample is put in the strip groove of strip frame 2;
4. keyboard reads in detected parameters, comprise on the IC-card 19 that the checking matter examination criteria working curve of storage and quality control band optical density reference monitor value, System self-test sweep time, sample title, sample number into spectrum, strip lot number, strip imitate phase, tester's name, detect date, IC-card password etc., system begins the self check timing automatically afterwards;
5. System self-test finishes, data processing equipment 15 reads IC-card 19 parameters, and drive 2 motions of strip frame to be fit to fibre-optical probe 8, the 9 pairs of strips 1 detect be with 26 and quality control band 27 carry out autoscan, the light signal that scanning obtains is after cmos image sensor 13 carries out photoelectricity and mould/number conversion, be converted to digital signal, be transferred to data processing equipment 15;
6. the optical density value OD of checking matter characteristic wave strong point in the data processing equipment 15 automatic discriminating digit signals
Detect bandWith quality control band optical density value OD
Quality control band, calculate OD automatically
Detect band/ OD
Quality control bandRatio or OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio, and according in the IC-card 19 storage corresponding checking matter examination criteria working curve calculate checking matter concentration automatically, the quality control band optical density reference monitor value (OD that IC-card 19 provides simultaneously
Quality control band) can judge whether gained checking matter concentration is effective;
7. analysis result is transferred to output display device 16, the output result.
The technical problems to be solved in the utility model
First problem to be solved in the utility model provides a kind of quantitative detection system of quantum dot mark test strip based on cmos image sensor.This detection system can not only be carried out single or mixes quantitatively or qualitative detection the target checking matter, and can also realize immunochromatography reaction dynamic process is observed.Described detection system has detection sensitivity height, testing result objective and fair, uses advantages such as flexible.
Second problem to be solved in the utility model provides the detection using method of described detection by quantitative system.
The utility model beneficial effect
(1) can realize quick and precisely that sample is single or polycomponent quantitative, qualitative detection.Because different-grain diameter, the fluorescence excitation that the quantum dot of composition and structure produces is a specific wavelength fluorescence, and the quantum dot potpourri can not produce the variable fluorescence of spectrum, the utility model is selected different-grain diameter for use, the quantum dot-labeled different target of composition and structure detects thing and makes quanta dot mark test bar, with described system the individual features wavelength fluorescent that the quantum dot-labeled compound of strip produces is carried out interpretation, therefore can realize accurately that sample is single or polycomponent is quantitative, qualitative detection: output display device 16 can quantitatively show single or the accurate concentration of polycomponent checking matter by being used host computer or alphanumeric LCD; Output display device 16 can the single or polycomponent checking matter testing result of qualitative demonstration by being used LED or sound.
(2) can carry out dynamic monitoring to course of reaction.The utility model detects the OD of band and quality control band according to strip
Detect band/ OD
Quality Control BandRatio or OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio, utilize with joining on the IC-card 19 each target checking matter respective standard working curve cause data processing equipment 15 of storage and analyze automatically and determine target checking matter concentration.If the detection reaction failure, resulting quality control band optical density value OD in the testing process
Quality control bandUnderstand quality control band optical density reference monitor value OD with IC-card 19 storages
Quality control bandThere is very big statistical errors, prompting strip reaction failure.Typical curve on the IC-card 19 and quality control band optical density reference monitor value can be adjusted with the strip differences between batches, have overcome different batches, different manufacturers and have produced the checking matter concentration detection difference that the strip mass discrepancy may cause; The utility model has also carried out investigating repeatedly to the strip reaction time before the formal test sample of system, this strip reaction time is fitted to System self-test and also is identified on the IC-card 19 sweep time in the utility model, but this time parameter order set just began formally sample to be detected after this time, guaranteed the strip reactant before system formally begins to detect, fully ooze move to strip detect be with 26 and quality control band 27 positions to be fit to fibre-optical probe 8, the 9 pairs of detections be with 26 and quality control band 27 carry out autoscan, improved the accuracy that detects.
(3) can realize the online detection of sample.Formerly technology (number of patent application 200510059892.3) was used for the photoelectron test macro that the quantum dot strip detects though once disclose a kind of comprising, but this patent adopts photomultiplier to make photodetector, can not realize the online detection of sample, limited this formerly technology use at vast outdoor location.System described in the utility model adopts cmos image sensor, is not only applicable to indoor detection, but also can realizes the online detection of sample, and vast outdoor location all can conveniently use.
(4) can make things convenient for tester user that testing result information is carried out flexible real time access and taken outward.IC-card 19 described in the utility model links to each other with data processing equipment 15 by the USB serial ports, IC-card 19 is except that being used to store checking matter respective standard working curve and the quality control band optical density reference monitor value, also can be used for storing tester's sample detection object information temporarily, the tester can plug IC-card 19 in real time according to own needs, thus make things convenient for the tester realize flexibly to the access of testing result information with take outward.
(5) detect fast, easy to operate, detecting does not need other instrument and equipments and reagent, as long as after the tester will be added with the quanta dot mark test bar imbedding system of sample, a few minutes just can accurately obtain testing result.Testing result is objective, and is highly sensitive.
Description of drawings
Fig. 1: based on the quantitative detection system of quantum dot mark test strip structured flowchart of cmos image sensor.
Fig. 2: strip side-looking structural drawing.
Fig. 3: strip plan structure figure.
Fig. 4: hepatitis B virus surface antigen (HBsAg) examination criteria working curve.
Fig. 5: quantum dot-labeled HBsAg fluorescent spectrum curve.
Fig. 6: tumor markers AFP, CEA, PSA examination criteria working curve.
Fig. 7: quantum dot-labeled AFP, CEA, PSA fluorescent spectrum curve.
Sequence number is expressed as follows: 1, strip; 2, strip frame; 3, scanning system; 4, light source; 5, incident light coupling mechanism; 6, incident optical; 7, incident optical; 8, fibre-optical probe; 9, fibre-optical probe; 10, outgoing optical fiber; 11, outgoing optical fiber; 12, emergent light coupling mechanism; 13, cmos image sensor; 14, well (its down corresponding sample pad 20); 15, data processing equipment; 16, output display device (as host computer, alphanumeric LCD, LED, sound etc.); 17, printer; 18, keyboard; 19, IC-card; 20, sample pad; 21, glass fibre membrane pad; 22, analyzing film; 23, strong adsorptive pads; 24, strip reaction end indicating label; 25, strip identification label; 26, detect band; Outgoing optical fiber, 30, driver module that the incident optical that 27 quality control bands, 28 fibre-optical probes (cross section sight) are located, 29 fibre-optical probes (cross section sight) are located; 31, stepper motor
Embodiment
The utility model is described in further detail below in conjunction with drawings and Examples, but should not limit protection domain of the present utility model with this.
(1) quantitative detection system of quantum dot mark test strip structure described in the utility model
Fig. 1 is the quantitative detection system of quantum dot mark test strip structured flowchart based on cmos image sensor described in the utility model.Described system comprises quanta dot mark test bar 1, is used to place the supporting IC-card 19 of the strip frame 2, scanning system 3, cmos image sensor 13, data processing equipment 15, output display device 16, printer 17, keyboard 18 of strip and and strip 1.
The strip frame 2 of described system has the strip groove.The strip groove is used to place quanta dot mark test bar to be measured 1.Described strip 1 is provided with sample pad 20, the glass fibre membrane pad 21 that is coated with quantum dot-labeled thing that mutual overlap joint pastes in turn, have and detect with 26 and the analyzing film 22 of quality control band 27, strong adsorptive pads 23, strip reaction end indicating label 24 and strip identification label 25.
Described scanning system 3 comprises an excitation source 4, is followed successively by incident light coupling mechanism 5, incident optical 6,7, fibre-optical probe 8,9, outgoing optical fiber 10,11, emergent light coupling mechanism 12 on the output light path of this excitation source 4, until cmos image sensor 13.Light source 4 comprises LED or laser diode.Fibre-optical probe 8,9 closes Shu Gongyong, and its center is an incident optical 28, and periphery is an outgoing optical fiber 29.Incident optical 6, bundle was through fibre-optical probe 8 in 7 minutes two, 9 shine respectively strip detect with 26 and quality control band 27 on the quantum dot-labeled compound of excitation-detection band 26 and quality control band 27 send the characteristic wavelength reflected fluorescent light, the corresponding outgoing optical fiber 10,11 with quality control band of band, emergent light coupling mechanism 12 backs are received by cmos image sensor 13 two bundle reflected fluorescent light after testing respectively.
The signal output part of described cmos image sensor 13 links to each other with the signal input part of data processing equipment 15.Cmos image sensor 13 be used for detection with 26 and the quality control band 27 transmission quantum dot-labeled compound characteristic wavelength reflected fluorescence signal of coming carry out photoelectricity and mould/number conversion, thereby become digital signal.
Described data processing equipment 15 is gathered, the next digital signal of storage cmos image sensor 13 transmission, and digital signal is carried out data processing.Described data processing equipment 15 also is furnished with output display device 16, printer 17, keyboard 18 and an IC-card that matches with strip 19, and they link to each other with data processing equipment 15 respectively.Also has a Drive and Control Circuit 30 between described data processing equipment 15 and the strip frame 2,31, this circuit 30, thereby 31 can make described detection by quantitative system by data processing equipment 15 read send after the parameter on the IC-card 19 instruction control automatically 2 motions of strip frame with 8, the 9 pairs of strips of fibre-optical probe that are fit to scanning system 3 detect be with 26 and quality control band 27 carry out autoscan.Described data processing equipment 15 can corresponding data be handled and single-chip microcomputer, CPU or the PC of Control Software in order to have.
The output display device 16 of described system can be host computer, alphanumeric LCD, LED, sound etc.Described system cooperates different output display devices 16, realizes the quantitative or qualitative detection of sample single component or polycomponent.That is: when described system was used for the sample detection by quantitative, output display device 16 can be host computer or alphanumeric LCD; When described system was used for the sample qualitative detection, output display device 16 can be LED or sound etc.
Adopt the USB serial communication between described IC-card 19 and the data processing equipment 15.During sample detection, IC-card 19 can plug in real time.IC-card 19 is stored each checking matter respective detection standard working curve and strip quality control band optical density reference monitor value.Each checking matter respective detection standard working curve that IC-card 19 is stored can adopt one of following two kinds of curve modes: the ratio (OD of one detection band optical density value/quality control band optical density value that to be checking matter standard items series concentration record through quantitative system described in the utility model with the serial concentration of this checking matter standard items
Detect band/ OD
Quality control band) between the corresponding relation curve; Also ratio (the OD of detection band optical density value/(detect and be with optical density value+quality control band optical density value) that can record through detection by quantitative described in the utility model system for checking matter standard items series concentration and this checking matter standard items series concentration
Detect band/ (OD
Detect band+ OD
Quality control band)) between the corresponding relation curve.The strip quality control band 26 optical density value (OD that obtain during standard curve making
Quality control band) be stored in simultaneously the reference monitor value of double as strip biological respinse success or failure on the IC-card 19 with standard working curve.In the sample actual detected, if the reaction failure, the OD that sample detection obtains
Quality control bandThe corresponding OD that stores on value and the IC-card 19
Quality control bandValue will produce very big statistical errors.IC-card 19 is stored parameter and is matched with each batch strip product.During sample detection, the operator inserts IC-card 19, data processing equipment 15 sends the fibre-optical probe 8 that 2 motions of the automatic control of instruction strip frame make scanning system 3 after reading IC-card 19 parameters, the 9 pairs of strips 1 detect be with 26 and quality control band 27 carry out autoscan, thereby obtain detecting with 26 and the characteristic frequency fluorescence optical density value sent of quality control band 27, and can realize that according to the corresponding checking matter standard working curve (simultaneously with reference to the suggested biological respinse success or failure of strip quality control band optical density reference monitor value) of IC-card 19 storages checking matter concentration accurately measures.IC-card 19 also can be made temporary storage storage testing result information, and each detection can be printed or preserve.
The method for making of each checking matter respective standard working curve of described IC-card 19 storages is: (1) preparation standard items series concentration; (2) each standard items series concentration is placed detection by quantitative described in the utility model system record corresponding OD
Detect bandAnd OD
Quality control bandAnd calculate OD respectively
Detect band/ OD
Quality control bandRatio or OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio; (3) make X-axis with each standard items series concentration, test result is with OD
Detect band/ OD
Quality control bandRatio is made Y-axis, the drawing standard working curve; Or: make X-axis with each standard items series concentration, test result is with OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio makes Y-axis, the drawing standard working curve; (4) write standard working curve software, and this Curve Software and strip quality control band optical density reference monitor value are stored on the IC-card 19.
One synchronous control circuit (not shown) can also be arranged to avoid 13 integral time of cmos image sensor long and cause noise level to improve between the light source 4 of described system and the cmos image sensor 13.
The utility model adopts strip with joining the IC-card 19 corresponding checking matter standard working curves of storage and providing strip quality control band optical density reference monitor value prompting reaction success or failure way by IC-card 19.Typical curve on the IC-card 19 and quality control band optical density reference monitor value can be adjusted with the strip differences between batches.The utility model adopts in 3 minutes later particular moments of described detection by quantitative system boot and just formally makes described system begin sample detection, to be fit to the work of system's IC-card examination criteria working curve and strip quality control band optical density reference monitor value.This strip reaction time is fitted to System self-test sweep time in the utility model, its parameter still is identified on the IC-card 19 that matches with strip.
The quantum dot of quanta dot mark test bar 1 described in the utility model is a semiconductor nano material, comprising: ZnS, CdS, HgS, ZnSe, CdSe, HgSe, CdTe, ZnTe, ZnO, PbSe, HgTe, GaAs, InP, InAs, InCaAs, CdS/ZnS, CdS/Ag
2S, CdS/PbS, CdS/Cd (OH)
2CdS/HgS, CdS/HgS/CdS, ZnS/CdS, ZnS/CdS/ZnS, ZnS/HgS/ZnS/CdS, CdSe/CdS, CdSe/ZnS, CdSe/ZnSe, CdSe/CuSe, CdSe/HgTe, CdSe/HgSe, CdSe/HgSe/CdSe, CdTe/HgS, CdTe/HgTe, InAs/InP, InAs/CdSe, InAs/ZnSe, MgS, MgSe, MgTe, CaS, CaSe, CaTe, SrS, SrSe, SeTe, BaS, BaSe, BaTe, CdS:Mn, ZnS:Mn, CdS:Cu, ZnS:Cu, CdS:Tb, the combination of any one among the ZnS:Tb or any several nano particles, and be nuclear by above-mentioned any one quantum dot, silicon dioxide is the core-shell type nano-complex particle of shell.Above-mentioned quantum dot-labeled target checking matter comprises plurality of target checking matters such as pathogen, antigen, antibody, illegal drug, major disease (tumour, cancer, angiocardiopathy and diabetes etc.), persticide residue, food security biological detection.The utility model carries out single to above-mentioned quantum dot-labeled target checking matter or mixes quantitative or qualitative detection.
Compared with prior art, characteristics of the present utility model are: can realize the accurate detection by quantitative of sample polycomponent, also can carry out sample sxemiquantitative or qualitative detection as required; Utilize fibre-optical probe 8,9 scanning strips detect be with 26 and quality control band 27 to excite strip quantum dot-labeled deposits yields characteristic wavelength fluorescence and in conjunction with strip reaction time (i.e. the described System self-test in start back sweep time), can conveniently analyze oozing of strip reactant and move distribution, realization reaches the success or failure details of understanding this bioprocesses in depth to the dynamic monitoring of bioprocesses; Process software (as IC-card) can organically combine automatically quanta dot mark test bar detect be with 26 and quality control band 27 fluorescence strong and weak accurately quantitatively or sxemiquantitative target checking matter concentration or qualitative detection target checking matter have or not, overcome the testing result difference that the strip product of different batches, different manufacturers production causes because of its mass discrepancy, testing result is objective, easy to operate, do not need other any instrument and equipment and reagent, just can accurately obtain testing result, high efficiency in several minutes.
(2) course of work of detection by quantitative described in the utility model system
The course of work of detection by quantitative described in the utility model system is: the quanta dot mark test bar to be measured 1 that will be added with sample is put in the strip groove of strip frame 2, data processing equipment 15 sends instruction and controls the fibre-optical probe 8 that 2 motions of strip frame make scanning system 3 automatically, the detection of 9 pairs of strips be with 26 and quality control band 27 carry out autoscan, the light that light source sends enters incident optical 6 through incident light coupling mechanism 5,7, incident optical 6, bundle was through fibre-optical probe 8 in 7 minutes two, 9 shine respectively strip detect with 26 and quality control band 27 on quantum dot-labeled compound on excitation-detection band 26 and the quality control band 27 send the characteristic wavelength reflected fluorescent light, two bundle reflected fluorescent light are with corresponding outgoing optical fiber 10 with quality control band respectively after testing, 11 and emergent light coupling mechanism 12 back received by cmos image sensor 13, after cmos image sensor 13 carries out photoelectricity and mould/number conversion with light signal, store and data processing for data processing equipment 15 digital data transmission after the conversion, data processing equipment 15 automatically to detection with 26 and the optical density value of the characteristic frequency fluorescence that comes of quality control band 27 transmission discern, obtain each checking matter and detect and be with 26 optical density value (OD
Detect band) and quality control band 27 optical density value (OD
Quality control band), and then calculate OD
Detect band/ OD
Quality control bandRatio or OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio, and calculate checking matter concentration according to the corresponding checking matter standard working curve analysis meter of storage on the IC-card 19 automatically, and afterwards testing result is transferred to output display device 16, finish whole testing process.
(3) the detection using method of detection by quantitative described in the utility model system
The detection using method of detection by quantitative described in the utility model system realizes by following step:
1. the energized of starting shooting;
2. insert the IC-card 19 that storage has checking matter examination criteria working curve and strip quality control band optical density reference monitor value;
3.) quanta dot mark test bar 1 that will be added with testing sample is put in the strip groove of strip frame 2;
4. keyboard reads in detected parameters, comprise on the IC-card 19 that the checking matter examination criteria working curve of storage and quality control band optical density reference monitor value, System self-test sweep time, sample title, sample number into spectrum, strip lot number, strip imitate phase, tester's name, detect date, IC-card password etc., system begins the self check timing automatically afterwards;
5. System self-test finishes, data processing equipment 15 reads IC-card 19 parameters, and drive 2 motions of strip frame to be fit to fibre-optical probe 8, the 9 pairs of strips 1 detect be with 26 and quality control band 27 carry out autoscan, the light signal that scanning obtains is after cmos image sensor 13 carries out photoelectricity and mould/number conversion, be converted to digital signal, be transferred to data processing equipment 15;
6. the optical density value OD of checking matter characteristic wave strong point in the data processing equipment 15 automatic discriminating digit signals
Detect bandWith quality control band optical density value OD
Quality control band, calculate OD automatically
Detect band/ OD
Quality control bandRatio or OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio, and according in the IC-card 19 storage corresponding checking matter examination criteria working curve calculate checking matter concentration automatically, the quality control band optical density reference monitor value (OD that IC-card 19 provides simultaneously
Quality control band) can judge whether gained checking matter concentration is effective.
7. analysis result is transferred to output display device 16, the output result.
The embodiment 2 detection by quantitative detection hepatitis B surface antigen HBsAg (individual event detection) of system described in the utility model
(1) quantum dot-labeled hepatitis B surface antigen HBsAg strip is made
The quanta dot mark test bar that described system detects usefulness is provided with sample pad 20, the glass fibre membrane pad 21 that is coated with quantum dot-labeled HBsAg monoclonal antibody that mutual overlap joint pastes in turn, have and detect with 26 and the analyzing film 22 of quality control band 27, strong adsorptive pads 23, strip reaction end indicating label 24 and " HBsAg " strip identification label 25.Detection is with 26 to be coated with the HBsAg monoclonal antibody, and quality control band 27 is coated with Quality Control thing sheep anti mouse IgM antibody or sheep anti-mouse igg antibody or rabbit anti-mouse IgM antibody or rabbit anti-mouse igg antibody.
1, each component preparation of strip
(1) sample pad: select cellulose membrane for use, be cut into 297 * 15mm film piece, put in the elongate slots, add confining liquid (pH=7.20.03mol/L phosphate buffer+5%BSA ,+0.1%Tween 20) soak at room temperature 30min.Take out the film piece, 37 ℃ of oven dry, fully drying for standby.
(2) glass fibre membrane pad: select glass fibre membrane for use, be cut into 297 * 10mm film piece, put in the elongate slots, add preprepared quantum dot-HBsAg monoclonal antibody bond solution thereon, take out the film piece, 37 ℃ of oven dry, fully drying for standby.
(3) analyzing film: select nitrocellulose filter for use, be cut into 297 * 25mm film piece, put in the elongate slots, from film piece base by down from last separated by a distance with some film instrument respectively specking 0.5-5mg/mlHBsAg monoclonal antibody do to detect and be with and specking 0.5-5mg/ml sheep anti mouse IgM antibody or sheep anti-mouse igg antibody or rabbit anti-mouse IgM antibody or rabbit anti-mouse igg antibody are made quality control band, afterwards with made film piece in 37 ℃ of oven dry, abundant drying for standby.
(4) strong adsorptive pads: select cellulose membrane for use, be cut into 297 * 30mm film piece, fully drying for standby with strong water sorption.
(5) strip reaction end indicating label: selecting color change interval for use is the accurate pH test paper of 5.0-9.0, is cut into the film piece with 297 * 5mm, fully drying for standby.
(6) strip is debated and is known label and select for use the label paper that is printed with " HBsAg " sign to be cut into the film piece with 297 * 5mm, fully drying for standby.
2, strip assembling
Each component of strip for preparing is debated the mutual in turn overlap joint of knowledge label 25 by sample pad 20, glass fibre membrane pad 21, analyzing film 22, strong adsorptive pads 23, strip reaction end indicating label 24, strip stick on the plastic back plate, and cut into the strip of certain specification.This strip can be packed in the shell, and is standby under drying condition.This shell can be plastics.
(2) hepatitis B surface antigen HBsAg examination criteria working curve is drawn and storage
Fig. 4 is a HBsAg examination criteria working curve.Its method for making is as follows:
1, HBsAg standard items series concentration preparation
Normal human serum (with the dilution of pH=7.20.03mol/L PB damping fluid) with dilution in 1: 10 is made dilution, the HBsAg standard items is made into 20 parts of series concentration: 0pg/ml, 100pg/ml, 200pg/ml, 300pg/ml, 400pg/ml, 500pg/ml, 600pg/ml, 700pg/ml, 800pg/ml, 900pg/ml, 1000pg/ml, 1100pg/ml, 1200pg/ml, 1300pg/ml, 1400pg/ml, 1500pg/ml, 1600pg/ml, 1700pg/ml, 1800pg/ml, 1900pg/ml.
2, draw HBsAg examination criteria working curve
Above-mentioned each HBsAg standard items series concentration is provided with under the condition in identical systems with 10 quantum dot-labeled HBsAg strips respectively and detects 10 times, reads to such an extent that it detects band optical density value (OD respectively
Detect band) and quality control band optical density value (OD
Quality control band), obtain mean value and OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio.Make X-axis with HBsAg standard items series concentration, the OD that tries to achieve with each HBsAg standard items series concentration correspondence
Detect band/ (OD
Detect band+ OD
Quality control band) ratio makes Y-axis drawing standard working curve, the results are shown in Figure 4.
3, write HBsAg examination criteria working curve software, with this Curve Software together with OD
Quality control bandValue (in the sample actual detected, this OD
Quality control bandValue is as strip quality control band optical density reference monitor value) deposit IC-card in.
(3) hepatitis B patient blood serum sample HBsAg detects
1, sample source: the hepatitis B patient blood serum sample is provided by certain healthcare hospital for women ﹠ children, and blood serum sample is done 10 times of dilutions with pH=7.20.03mol/L PB damping fluid before detecting.
2, sample detection:
(1) system power supply is connected in start;
(2) insert the IC-card 19 that storage has HBsAg examination criteria working curve and strip quality control band optical density reference monitor value;
(3) quanta dot mark test bar 1 that will be added with hepatitis B patient test serum sample is put in the strip groove of strip frame 2;
(4) keyboard reads in detected parameters: comprise the HBsAg standard working curve of IC-card and quality control band optical density reference monitor value, inspection product title, inspection product numbering, strip lot number, strip effect phase, detection date, tester's name, IC-card password etc., System self-test was made as 5 minutes sweep time, and system begins the self check timing automatically afterwards;
(5) System self-test finishes, data processing equipment 15 reads IC-card 19 parameters, and drive 2 motions of strip frame to be fit to fibre-optical probe 8, the 9 pairs of strips 1 detect be with 26 and quality control band 27 carry out autoscan, the light signal that scanning obtains is after cmos image sensor 13 carries out photoelectricity and mould/number conversion, be converted to digital signal, be transferred to data processing equipment 15;
(6) detect the optical density value (OD that is with the characteristic wave strong point in the data processing equipment 15 automatic discriminating digit signals
Detect band) and quality control band optical density value (OD
Quality control band), calculate OD automatically
Detect band/ (OD
Detect band+ OD
Quality control band) ratio, and calculate HBsAg concentration in the blood serum sample automatically according to the HBsAg standard working curve that stores in the IC-card.As the present embodiment blood serum sample through as described in the detection by quantitative system to detect its HBsAg concentration be 205.9563pg/ml;
(7) output display device 16 shows testing result;
(8) system print test results report.
3, hepatitis B patient blood serum sample HBsAg ultimate density is calculated:
The HBsAg concentration x serum diluting multiple that HBsAg ultimate density (pg/ml)=system records in the blood serum sample
In the present embodiment, described blood serum sample HBsAg ultimate density=205.9563pg/ml * 10=2059.563pg/ml
Fig. 5 is quantum dot-labeled HBsAg fluorescent spectrum curve.Wherein, I is for detecting the HBsAg spectrum peak in the band, and II is the Quality Control thing spectrum peak in the quality control band.
The embodiment 3 detection by quantitative detection blood serum tumor markers AFP of system described in the utility model, CEA and PSA (inspection is multinomial)
(1) the quantum dot-labeled multinomial strip of tumor markers-inspection is made
The multinomial strip of described quantum dot-labeled tumor markers one inspection is provided with sample pad 20, glass fibre membrane pad 21 that mutual overlap joint pastes in turn, have and detect with 26 and the analyzing film 22 of quality control band 27, strong adsorptive pads 23, strip reaction end indicating label 24 and " AFP/CEA/PSA " strip identification label 25.Glass fibre membrane pad 21 is coated with the potpourri of quantum dot-labeled alpha-fetoprotein (AFP) monoclonal antibody, quantum dot-labeled carcinomebryonic antigen (CEA) monoclonal antibody and quantum dot-labeled prostate specific antigen (PSA) monoclonal antibody; Detection is with 26 potpourris that are coated with AFP antibody, CEA antibody and PSA antibody; Quality control band 27 is coated with Quality Control thing sheep anti mouse IgM antibody or sheep anti-mouse igg antibody or rabbit anti-mouse IgM antibody or rabbit anti-mouse igg antibody.
1, each component preparation of strip
(1) sample pad: select cellulose membrane for use, be cut into 297 * 15mm film piece, put in the elongate slots, add confining liquid (pH=7.20.03mol/L phosphate buffer+5%BSA ,+0.1%Tween 20) soak at room temperature 30min.Take out the film piece, 37 ℃ of oven dry, fully drying for standby.
(2) glass fibre membrane pad: select glass fibre membrane for use, be cut into 297 * 10mm film piece, put in the elongate slots, add preprepared quantum dot-labeled thing solution (for containing the mixture solution of quantum dot-labeled AFP monoclonal antibody, quantum dot-labeled CEA monoclonal antibody, quantum dot-labeled PSA monoclonal antibody) thereon, take out the film piece, 37 ℃ of oven dry, fully drying for standby.
(3) analyzing film: select nitrocellulose filter for use, be cut into 297 * 25mm film piece, put in the elongate slots, be with by following the detection from film piece base from last potpourri work with some film instrument specking AFP antibody (0.5-5mg/ml), CEA antibody (0.5-5mg/ml) and PSA antibody (0.5-5mg/ml) separated by a distance, make quality control band with a film instrument specking 0.5-5mg/ml sheep anti mouse IgM antibody or sheep anti-mouse igg antibody or rabbit anti-mouse IgM antibody or rabbit anti-mouse igg antibody, afterwards with made film piece in 37 ℃ of oven dry, abundant drying for standby.
(4) strong adsorptive pads: select cellulose membrane for use, be cut into 297 * 30mm film piece, fully drying for standby with strong water sorption.
(5) strip reaction end indicating label: selecting color change interval for use is the accurate pH test paper of 5.0-9.0, is cut into the film piece with 297 * 5mm, fully drying for standby.
(6) strip is debated and is known label and select for use the label paper that is printed with " AFP/CEA/PSA " sign to be cut into the film piece with 297 * 5mm, fully drying for standby.
2, strip assembling
Each component of strip for preparing is debated the mutual in turn overlap joint of knowledge label 25 by sample pad 20, glass fibre membrane pad 21, analyzing film 22, strong adsorptive pads 23, strip reaction end indicating label 24, strip stick on the plastic back plate, and cut into the strip of certain specification.This strip can be packed in the shell, and is standby under drying condition.This shell can be plastics.
(2) standard working curve is drawn and storage
Fig. 6 is AFP, CEA, PSA examination criteria working curve.Its method for making is as follows:
1, standard items series concentration preparation
Normal human serum (with the dilution of pH=7.2 0.03mol/L PB damping fluid) with dilution in 1: 10 is made dilution, and AFP, CEA and PSA standard items are made into each 20 parts of series concentration by 0pg/ml, 100pg/ml, 200pg/ml, 300pg/ml, 400pg/ml, 500pg/ml, 600pg/ml, 700pg/ml, 800pg/ml, 900pg/ml, 1000pg/ml, 1100pg/ml, 1200pg/ml, 1300pg/ml, 1400pg/ml, 1500pg/ml, 1600pg/ml, 1700pg/ml, 1800pg/ml, 1900pg/ml respectively.
2, draw tumor markers examination criteria working curve
Every part of tumor markers standard items series concentration is provided with under the condition in identical systems with the multinomial strip of 10 quantum dot-labeled tumor markerses, one inspection respectively and detects 10 times, reads to such an extent that it detects band optical density value (OD respectively
Detect band) and quality control band optical density value (OD
Quality control band), obtain mean value and OD
Detect band/ OD
Quality control bandRatio.Make X-axis with each tumor markers standard items series concentration, the OD that tries to achieve with each tumor markers standard items series concentration correspondence
Detect band/ OD
Quality control bandRatio is made Y-axis drawing standard working curve, the results are shown in Figure 6.
3, write tumor markers examination criteria working curve software, with this Curve Software together with OD
Quality control bandValue (in the sample actual detected, this OD
Quality control bandValue is as strip quality control band optical density reference monitor value) deposit IC-card in.
(3) tumor patient blood serum sample tumor markers detects
1, sample source: blood serum sample is provided by certain tumour hospital laboratory.Blood serum sample is done 10 times of dilutions with pH=7.2 0.03mol/LPB damping fluid before detecting.
2, sample detection:
(1) system power supply is connected in start;
(2) insert the IC-card 19 that storage has AFP, CEA, PSA examination criteria working curve and strip quality control band optical density reference monitor value;
(3) quanta dot mark test bar 1 that will be added with tumor patient test serum sample is put in the strip groove of strip frame 2;
(4) keyboard reads in detected parameters: AFP, CEA, PSA standard working curve and the quality control band optical density reference monitor value, inspection product title, inspection product numbering, strip lot number, the strip that comprise IC-card are imitated the phase, are detected date, tester's name, IC-card password etc., System self-test was made as 5 minutes sweep time, and system begins the self check timing automatically afterwards;
(5) System self-test finishes, data processing equipment 15 reads IC-card 19 parameters, and 2 motions of driving strip frame make fibre-optical probe 8, the 9 pairs of strips 1 detect be with 26 and quality control band 27 carry out autoscan, the light signal that scanning obtains is after cmos image sensor 13 carries out photoelectricity and mould/number conversion, be converted to digital signal, be transferred to data processing equipment 15;
(6) detect the optical density value (OD that is with the characteristic wave strong point in the data processing equipment 15 automatic discriminating digit signals
Detect Band) and quality control band optical density value (OD
Quality control band), calculate OD automatically
Detect band/ OD
Quality control bandRatio, and calculate AFP concentration in the blood serum sample automatically according to the AFP that stores in the IC-card, CEA, PSA standard working curve.As the present embodiment blood serum sample through as described in the detection by quantitative system to detect its AFP concentration be that 1753.4526pg/ml, CEA concentration are that 878.3892pg/ml, PSA concentration are 983.4257pg/ml;
(7) output display device 16 shows testing result;
(8) system print test results report.
3, tumor patient blood serum sample AFP, CEA, PSA ultimate density are calculated:
Formula is: blood serum sample tumor markers ultimate density (pg/ml)=system records concentration * serum diluting multiple
Thus: in the present embodiment,
Blood serum sample AFP ultimate density=1753.4526pg/ml * 10=17534.526pg/ml
Blood serum sample CEA ultimate density=878.3892pg/ml * 10=8783.892pg/ml
Blood serum sample PSA ultimate density=983.4257pg/ml * 10=9834.257pg/ml
Fig. 7 is quantum dot-labeled AFP, CEA, PSA fluorescent spectrum curve.Wherein, I is for detecting the AFP spectrum peak in the band, and II is for detecting the CEA spectrum peak in the band, and III is for detecting the PSA spectrum peak in the band, and IV is the Quality Control thing spectrum peak in the quality control band.
Claims (6)
1. quantitative detection system of quantum dot mark test strip based on cmos image sensor is characterized in that: it comprises quanta dot mark test bar (1), is used to place the strip frame (2) of strip, scanning system (3), cmos image sensor (13), data processing equipment (15), output display device (16), printer (17), keyboard (18) and one and the supporting IC-card (19) of strip:
Described scanning system (3) comprises an excitation source (4), on the output light path of this excitation source (4), be followed successively by incident light coupling mechanism (5), incident optical (6,7), fibre-optical probe (8,9), outgoing optical fiber (10,11), emergent light coupling mechanism (12), until cmos image sensor (13);
The signal output part of cmos image sensor (13) links to each other with the signal input part of data processing equipment (15), and data processing equipment (15) links to each other with output display device (16), printer (17), keyboard (18) and IC-card (19) respectively;
Between described data processing equipment (15) and the strip frame (2) Drive and Control Circuit (30 is arranged, 31), described data processing equipment (15) reads IC-card (19) parameter and sends instruction by this Drive and Control Circuit (30,31) control strip frame (2) motion automatically and the detection band (26) and the quality control band (27) of strip (1) carried out autoscan with the fibre-optical probe (8,9) that is fit to scanning system (3);
Described IC-card (19) is stored checking matter examination criteria working curve and strip quality control band optical density reference monitor value;
The light that light source (4) sends enters incident optical (6 through incident light coupling mechanism (5), 7), incident optical (6,7) divide two bundles through fibre-optical probe (8,9) the quantum dot-labeled compound that shines last excitation-detection band (26) of strip detection band (26) and quality control band (27) and quality control band (27) respectively sends the characteristic wavelength reflected fluorescent light, two bundle reflected fluorescence signal are with corresponding outgoing optical fiber (10 with quality control band respectively after testing, 11), emergent light coupling mechanism (12) is received by cmos image sensor (13), cmos image sensor (13) carries out photoelectricity and mould/number conversion with light signal, digital data transmission after the conversion stores and data processing for data processing equipment (15), data processing equipment (15) is discerned automatically to the characteristic frequency fluorescence optical density that detects band (26) and quality control band (27), and carry out checking matter concentration according to the checking matter standard working curve that IC-card (19) stores and calculate and technical Analysis, give output display device (16) with result transmission.
2. the quantitative detection system of quantum dot mark test strip based on cmos image sensor according to claim 1 is characterized in that: described excitation source (4) comprises LED or laser diode.
3. the quantitative detection system of quantum dot mark test strip based on cmos image sensor according to claim 1 is characterized in that: described data processing equipment (15) can corresponding data be handled and single-chip microcomputer, CPU or the PC of Control Software in order to have.
4. the quantitative detection system of quantum dot mark test strip based on cmos image sensor according to claim 1 is characterized in that: described output display device (16) can be host computer, alphanumeric LCD, LED or sound.
5. the quantitative detection system of quantum dot mark test strip based on cmos image sensor according to claim 1, it is characterized in that: described quanta dot mark test bar is the quanta dot mark test bar that described detection by quantitative system matches, be provided with sample pad (20), glass fibre membrane pad (21), analyzing film (22), strong adsorptive pads (23), strip reaction end indicating label (24) and strip identification label (25) that mutual overlap joint is pasted on it in turn with detection band (26) and quality control band (27).
6. the quantitative detection system of quantum dot mark test strip based on cmos image sensor according to claim 5, it is characterized in that: the glass fibre membrane pad (21) of described quanta dot mark test bar is coated with the quantum dot-labeled associated biomolecule molecule that the target checking matter detects that is used for, detect band (26) and be coated with the relevant special biomolecule that is used for the detection of target checking matter, quality control band (27) is coated with and comprises two anti-Quality Control things.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009200073261U CN201535749U (en) | 2008-07-14 | 2009-02-11 | Quantum dot mark test strip quantitative detection system based on CMOS picture sensor |
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| CN200820064223.4 | 2008-07-14 | ||
| CN2009200073261U CN201535749U (en) | 2008-07-14 | 2009-02-11 | Quantum dot mark test strip quantitative detection system based on CMOS picture sensor |
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| CN201535749U true CN201535749U (en) | 2010-07-28 |
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|---|---|
| CN (1) | CN201535749U (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102565386A (en) * | 2011-12-29 | 2012-07-11 | 北京康美天鸿生物科技有限公司 | Magnetic fluorescent microsphere immunochromatography quantitative detection method |
| WO2014176753A1 (en) * | 2013-04-30 | 2014-11-06 | 成都领御生物技术有限公司 | Quantum-dot immunochromatographic test strip detection system and use thereof |
| WO2015070704A1 (en) * | 2013-11-13 | 2015-05-21 | 成都领御生物技术有限公司 | Test bar card |
| WO2015070700A1 (en) * | 2013-11-12 | 2015-05-21 | 成都领御生物技术有限公司 | Quantum dot-labeled test strip card |
| WO2015070699A1 (en) * | 2013-11-12 | 2015-05-21 | 成都领御生物技术有限公司 | Quantum dot-labeled test strip card |
| CN106153899A (en) * | 2015-03-24 | 2016-11-23 | 北京万泰生物药业股份有限公司 | For detecting the device of lateral flow Multiple detection reagent |
| CN107064022A (en) * | 2017-04-20 | 2017-08-18 | 陕西科技大学 | A kind of luminescent spectrum for measuring quantum dot varies with temperature the device and method of rule |
| CN107861387A (en) * | 2017-11-13 | 2018-03-30 | 浙江大学 | A kind of internal thermally coupled air separation column control device based on concentration curve optimized algorithm |
-
2009
- 2009-02-11 CN CN2009200073261U patent/CN201535749U/en not_active Expired - Lifetime
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102565386A (en) * | 2011-12-29 | 2012-07-11 | 北京康美天鸿生物科技有限公司 | Magnetic fluorescent microsphere immunochromatography quantitative detection method |
| CN102565386B (en) * | 2011-12-29 | 2014-11-05 | 北京康美天鸿生物科技有限公司 | Magnetic fluorescent microsphere immunochromatography quantitative detection method |
| WO2014176753A1 (en) * | 2013-04-30 | 2014-11-06 | 成都领御生物技术有限公司 | Quantum-dot immunochromatographic test strip detection system and use thereof |
| WO2015070700A1 (en) * | 2013-11-12 | 2015-05-21 | 成都领御生物技术有限公司 | Quantum dot-labeled test strip card |
| WO2015070699A1 (en) * | 2013-11-12 | 2015-05-21 | 成都领御生物技术有限公司 | Quantum dot-labeled test strip card |
| WO2015070704A1 (en) * | 2013-11-13 | 2015-05-21 | 成都领御生物技术有限公司 | Test bar card |
| CN106153899A (en) * | 2015-03-24 | 2016-11-23 | 北京万泰生物药业股份有限公司 | For detecting the device of lateral flow Multiple detection reagent |
| CN107064022A (en) * | 2017-04-20 | 2017-08-18 | 陕西科技大学 | A kind of luminescent spectrum for measuring quantum dot varies with temperature the device and method of rule |
| CN107861387A (en) * | 2017-11-13 | 2018-03-30 | 浙江大学 | A kind of internal thermally coupled air separation column control device based on concentration curve optimized algorithm |
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