CN101592659B - System and method for quantitative detection of test strips on basis of continuous fluorescent-substance markers - Google Patents
System and method for quantitative detection of test strips on basis of continuous fluorescent-substance markers Download PDFInfo
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Abstract
The invention belongs to the field of bio-medical instruments, and in particular relates to a system and a method for quantitative detection of test strips on the basis of continuous fluorescent-substance markers. The system comprises a continuous fluorescent-substance marker test strip, a test strip frame, a lighting system, an imaging system, a fluorescent image receiver, a signal amplifier, an analog/digital converter, a data processing-controlling system, an output display device, a printer, a keyboard and an IC card matched with the test strip. The data processing-controlling system reads parameters of the IC card and then controls the test strip frame to move so as to ensure that the light emitted by the lighting system is reflected via a dichroic mirror and then automatically scans the test strip; acquired characteristic wavelength reflection fluorescence is transmitted to the data processing-controlling system for optical density identification and concentration calculation via the fluorescent image receiver, the signal amplifier and the analog/digital converter; and the output display device displays results. The invention can quickly and accurately realize the quantitative or qualitative detection of single-component and multi-component samples. The system has the characteristics of high detection sensitivity, objective results, flexible use and the like.
Description
Technical field
The present invention relates to the biologic medical instrument field, be specifically related to a kind of based on the test strip quantitative detection system and the method thereof that continue fluorescent-substance markers.This system can carry out autoscan to the strip that continues fluorescent-substance markers 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 (RIA), 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.The laser-Induced Fluorescence Detection technology is considered to one of the sensitiveest at present detection method.This method has plurality of advantages such as highly sensitive, that selectivity good, response speed is fast, and its range of application spreads all over each field of biomedical check.
The fluorescent marker that the laser-Induced Fluorescence Detection technology is the most often used at present is organic fluorescent dye (like luciferin, rhodamine etc.).Organic fluorescence label fluorescence lifetime is short, is difficult to overcome back of the body amount fluorescence interference, and detection sensitivity is low.Stablize by force though the green fluorescent protein that occurs thereafter is luminous, big owing to the protein molecular volume, relatively more responsive to environment, practical operation is inconvenient.The quantum dot that occurs in recent years can be avoided solvent with the nano particle that contains a large amount of fluorescent reporter molecules (comprising quantum dot, rare earth chelate, organic dyestuff etc.) and disturb, and fluorescent emission is stablized by force, has become the new focus in immune labeled field at present.
Rare earth ion; Mainly be the lanthanide chelate of Eu (III), Tb (III), Sm (III), Dy (III), its excitation wavelength can be regulated the fluorescent emission high specificity through part; The Stokes displacement is big; The emitting fluorescence life-span is long, utilizes the time-resolved fluorescence technology can significantly reduce at the bottom of the biomedical sample copy and interference of stray light, and sensitivity can improve several magnitude.Utilization has rare earth ion Eu, Sm, Dy, the lanthanide chelate of Tb and the rare earth nanometer particle that their potpourri is formed of long-life fluorescence and thinks up-and-coming ultramicro-analysis method as fluorescent marker at present.
Quantum dot (Quantum Dot; Be called for short QD) be that a kind of of 20th century new development nineties has more the semiconductor nanocrystal of good spectral signature and photochemical stability with respect to conventional fluorescent dyestuff and lanthanide chelate, its fluorescence radiation efficient height, exciting line wide ranges, spectral line of emission narrow range, particle diameter and biomolecule is close, finishing multifunction, fluorescence lifetime are longer.As biological fluorescent labelling thing of new generation, aspects such as the interaction between its simultaneous determination of multiponents in immunoassay, immune tracer molecule location, nano biological sensor, detection in real time, cell imaging, in-vivo imaging, medical diagnosis on disease and research biomacromolecule, component circulation and the mode of action in body have unique effect and broad prospect of application.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 quantum dot major advantage 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 is high.Quantum dot is the polyelectron system, and luminescence efficiency is far above single organic molecule, and it is 10 in visible absorptivity with the 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 use the light deexcitation of several kinds of wavelength when measuring several kinds of fluorescence spectrums, and this analyzes for multiple labeling simultaneously and causes the deuce to pay.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) luminescent spectrum of big Stokes displacement of tool and 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 luciferin, and peak width at half height usually has only 40nm or littler, and does not have 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.
The architectural characteristic of quantum dot, rare earth chelate, Fluorescence Mechanism and long-life fluorescence characteristics; Make their when detecting, can effectively eliminate background fluorescence and disturb (comprise biosome autofluorescence disturb and analyze with the background fluorescence interference of material production etc.), thereby reach accurate test sample checking matter concentration purpose through a time delay as continuing fluorescent marker mark immunity-chromatography strip.And; Quantum dot is as continuing fluorescent marker; Owing to 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; Thereby can come that quantum dot-labeled target checking matter is carried out specific fluorescent according to the optical characteristics that the quantum dot of specific size, composition and structure produces specific fluorescent and measure, can reach the concentration of accurate test sample polycomponent object.
Adopt quanta dot mark test bar detection of biological medical sample; One Chinese patent application numbers 200610024086.7 has disclosed a kind of fast immune chromatographic strip detection method; The different quantum dot-labeled different target thing antibody of this application patent utilization; Under ultraviolet, can with the naked eye observe testing result fast, realize 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 instrument detecting 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 through naked eyes to judge testing result under ultraviolet; Testing result is subject to the influence of tester's experience; Subjectivity is strong, and 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 just difficulty judged result with the naked eye more 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.One 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.One 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 present invention.In addition; Author of the present invention has disclosed two kinds respectively based on quantum dot-labeled strip detection system and method thereof in one Chinese patent application number 200810186010.3,200820209876.7,200810045548.2,200820064223.4 files, but the structure of the optical imaging system in said detection system composition is different with the present invention.
At rare-earth fluorescent nanoparticle label strip aspect the detection of biological sample; One Chinese patent application numbers 200510052292.4 (denomination of invention: " a kind of with fluorescent rare earth nanometer particle serve as a mark thing immune chromatography method and detect strip ") disclosed a kind of immune chromatography method and detected strip; Same making to the rare earth nanometer particle mark test bar of this patented claim is described; The detection of its mark test bar need be by other fluoroscopic examination instruments; Observe or fluorescence measurement appearance through special use detects as with the naked eye detecting by Ultraviolet Detector or assisting with digital camera, this makes the practical application of said strip receive very big restriction.One Chinese patent application number 03819391.4 (denomination of invention: " adopting the detection based on film of time-resolved fluorescence ") is though disclosed a kind of pick-up unit based on the fluorescent rare earth nanometer particle label film, and this apparatus structure composition especially structure of its optical imaging system is formed different fully with the present invention.Also be pointed out that, use rare earth nanometer particle to make to continue the existing more bibliographical information of solid phase detection method of fluorescence labeling substance markers microwell plate at present, and developed corresponding supporting detector---time-resolved fluorescence detector (or system); Though its detection can reach higher sensitivity, can realize quantitatively or half-quantitative detection that shortcoming is a complicated operation; Relate to a plurality of steps such as application of sample, warm bath, washing, colour developing, termination, detection; Ability was accomplished in action need 2-4 hour, and operating personnel and environment are also had certain requirement, and; Except the expensive necessary instrument of needs---time resolution detector---; Also need wash other supplementary instrument equipment such as plate machine, therefore, its widespread usage still is restricted.
In view of the above-mentioned deficiency of prior art, the present invention discloses a kind of based on the test strip quantitative detection system and the method thereof that continue fluorescent-substance markers, and its strip making, said system architecture composition, detection method of application etc. are all different fully with prior art.
Detection by quantitative system according to the invention and detection method of application thereof are used for the quantitative or qualitative detection of biomedical sample; Fast simple; Easy to operate; Except that the lasting fluorescent-substance markers strip that uses with system support, hardware of IC-card and system own, no longer need other any instrument and equipment and reagent.Said strip is simple for production, and is with low cost, is disposable product.During sample detection, after the lasting fluorescence labeling strip that the tester only needs to be added with sample is inserted said system and inserted supporting IC-card, just can accurately obtain testing result in a few minutes.Testing result is objective, and is highly sensitive, can not only accurately realize the quantitative or qualitative detection of sample single component and polycomponent, can also carry out the dynamic monitoring prompting to strip immunochromatography reaction success or failure.
Summary of the invention
Technical scheme of the present invention is following:
Lasting fluorescent-substance markers test strip quantitative detection system according to the invention comprises and continues fluorescent-substance markers strip 18, the strip frame 28 that is used to place strip, illuminator, imaging system, fluoroscopic image receiver 8, signal amplifier 9, A/D converter 10, data processing and control system 11, output display device 12, printer 13, keyboard 14 and one and the supporting IC-card 15 of strip.
The strip frame 28 of said system has the strip groove.The strip groove is used to place lasting fluorescent-substance markers strip 18 to be measured.Said strip 18 is provided with sample pad 19 that mutual overlap joint pastes in order, be coated with the glass fibre membrane pad 20 that continues the fluorescent-substance markers thing, have and detect with 25 and the analyzing film 21 of quality control band 26, strong adsorptive pads 22, strip reaction end indicating label 23 and strip identification label 24.
Said illuminator comprises an excitation source 1, on the output light path of this excitation source 1, is followed successively by fibre bundle 2, collimated illumination lens 3, dichronic mirror 4, preceding mirror group 5, until lasting fluorescent-substance markers strip 18.Excitation source 1 comprises LED or laser diode.The light that fibre bundle 2 sends light source 1 is divided into and has a determining deviation and the identical two laser of intensity, this two laser by collimated illumination lens 3 collimations be incide behind the two bundle directional lights dichronic mirror 4 surfaces through preceding mirror group 5 shine synchronously strip detect with 25 with quality control band 26 on excite strip to detect to send the characteristic wavelength reflected fluorescent light with the lasting fluorescent-substance markers compound of quality control band 26 with 25.
Said imaging system comprises preceding mirror group 5, dichronic mirror 4, optical filter 6, the back mirror group 7 of common optical axis.Dichronic mirror 4 reflectings surface become miter angle with optical axis included angle.Preceding mirror group 5 all adopts isolating construction with back mirror group 7.Illuminator light path part before dichronic mirror 4 is perpendicular with the optical axis of said imaging system, part after dichronic mirror 4 and said imaging system common optical axis.
Said fluoroscopic image receiver 8 is positioned on mirror group 7 image planes of back.The signal output part of fluoroscopic image receiver 8 links to each other with A/D converter 10 through signal amplifier 9.Fluoroscopic image receiver 8 is collected strips and is detected the characteristic wavelength fluorescence signal that reflects with quality control band 26 with 25, converts thereof into to carrying out electric signal by signal amplifier 9 behind the electric signal and amplifies.Fluoroscopic image receiver 8 has multiple available object, comprises CCD, CMOS, photomultiplier, photodiode, phototriode.
The signal input part of described A/D converter 10 links to each other with fluoroscopic image receiver 8 signal amplifier 9 signal output parts afterwards, and the signal output part of A/D converter 10 links to each other with the signal input part of data processing with control system 11.Described A/D converter 10 is used for changing the amplification signal that signal amplifier 9 transmission come into digital signal.A conspicuous embodiment is; Said detection by quantitative system is when selecting for use CMOS to make fluoroscopic image receiver 8; Because the CMOS of commercial model has been integrated with function amplifier 9 and A/D converter 10 now; Therefore, the detection by quantitative system that constitutes thus is functions of use amplifier 9 and A/D converter 10 no longer in addition.
The strip that said data processing and control system 11 are gathered, 10 transmission of storage A/D converter come detects the digital signal of band and quality control band reflected fluorescence signal, and this digital signal is carried out data processing.Said data processing and control system 11 also are furnished with output display device 12, printer 13, keyboard 14 and an IC-card 15 that matches with strip, and they link to each other with control system 11 with data processing respectively.Also has a Drive and Control Circuit 16 between said data processing and control system 11 and the strip frame 28; 17; This circuit 16, thus 17 said detection by quantitative system is read through its data processing and control system 11 send instruction on the IC-card 15 after the parameter and control 28 motions of strip frame automatically and with the luminous energy that is fit to illuminator and sends strip is detected and be with 25 to carry out autoscan with quality control band 26.Said data processing and control system 11 can corresponding data be handled and single-chip microcomputer, CPU or the PC of Control Software in order to have.
The output display device 12 of said detection by quantitative system can be host computer, alphanumeric LCD, LED, audio unit etc.The different output display device 12 of said detection by quantitative system's cooperation; Can realize that sample single component or polycomponent are quantitatively or qualitative detection: when said detection by quantitative system was used for the sample detection by quantitative, output display device 12 can be host computer or alphanumeric LCD; When said detection by quantitative system was used for the sample qualitative detection, output display device 12 can be LED or audio unit etc.
Said IC-card 15 is stored has each checking matter respective detection standard working curve and strip quality control band optical density reference monitor value.Adopt the USB serial communication between IC-card 15 and data processing and the control system 11, during sample detection, IC-card 15 can plug in real time.Each checking matter respective detection standard working curve that IC-card 15 is stored can adopt one of following two kinds of curve modes: the ratio (OD of one of which detection band OD value/quality control band OD value that to be checking matter standard items series concentration record through detection by quantitative according to the invention system with the serial concentration of this checking matter standard items
Detect band/ OD
Quality control band) between the corresponding relation curve; Ratio (the OD of detection band OD value/(detect and be with OD value+quality control band OD value) that also can record through detection by quantitative according to the invention 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 28 OD 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 15 with standard working curve.In the sample actual detected, if the reaction failure, the OD that sample detection obtains
Quality control bandCorresponding OD with storage on the IC-card 15
Quality control bandValue will produce very big statistical errors.IC-card 15 stores parameter and each batch strip product matches.During sample detection; The operator inserts IC-card 15; Data processing and control system 11 read send after IC-card 15 parameters instruction automatically 28 motions of control strip frame make luminous energy that illuminator is sent and that reflect through dichronic mirror 4 to strip detect be with 25 and quality control band 26 carry out autoscan; Thereby obtain detecting the characteristic frequency fluorescence OD value that sends with quality control band 26 with 25, and can realize that according to the corresponding checking matter standard working curve (the biological respinse success or failure of being pointed out with reference to strip quality control band optical density reference monitor value simultaneously) of IC-card 15 storages checking matter concentration accurately measures.IC-card 15 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 said IC-card 15 storages is: (1) preparation standard items series concentration; (2) place detection by quantitative according to the invention system to record corresponding OD each standard items series concentration
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 the X axle with each standard items series concentration, test result is with OD
Detect band/ OD
Quality control bandRatio is made the Y axle, the drawing standard working curve, or: make the X axle with each standard items series concentration, test result is with OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio makes the Y axle, 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 15.
The present invention detect principle based on: continue the fluorescent-substance markers strip and detect and can send the individual features wavelength fluorescent after being excited by certain wavelength laser with the lasting fluorescent-substance markers compound on 25, 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 lasting fluorescent-substance markers compound on 25.Otherwise testing sample concentration is more little in the sample, strip detect the characteristic wavelength fluorescence signal that sends with the lasting fluorescent-substance markers compound on 25 just more a little less than.According to this relation; The present invention is applied to strip one end (being well 27 places) with testing sample; 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 22 of the strip other end to be moved; When target checking matter molecule (like checking matter specific antigen or antibody) is arranged in the fluid sample; They just the corresponding biomolecule that is labeled lasting fluorescence on the glass fibre membrane pad 20 that is coated on the preceding stage casing of strip combine and ooze the detection that moves to strip analyzing film 21 to be with 25 to the strip rear end; Detect band and continue the fluorescent-substance markers compound with being coated on to detect to combine to form with 25 special target checking matter correlation molecule (like the specific antibody or the antigen of respective objects checking matter molecule), a remaining part indicates the biomolecule that continues fluorescent marker and continues to ooze the Quality Control thing (anti-as two) that moves to strip quality control band 26 and encapsulate in advance with quality control band 26 and combines the lasting fluorescent-substance markers compound of formation quality control band.Be combined in strip detect with 25 with the lasting fluorescent-substance markers compound of quality control band 26 because of being combined with lasting fluorescence; They are under the exciting of the light that excitation source sends; Promptly launch corresponding long-life characteristic wavelength fluorescence, this characteristic wavelength continues fluorescence intensity can reflect checking matter concentration.Through receiving, change, transmit, handle this characteristic wavelength fluorescence intensity information, therefore can accurately detect checking matter concentration.
Lasting fluorescence according to the invention means and can continue to send the life-span material of longer characteristic wavelength fluorescence relatively after one type of certain wavelength excitation of quilt that it is particularly including quantum dot and nano rare earth fluorescence complex.
Said quantum dot comprises: 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)
2, any one or any several kinds of nano particles among the CdS/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, ZnS:Tb combination, and be that nuclear, silicon dioxide are the core-shell type nano-complex particle of shell by above-mentioned any one quantum dot.
Said nano rare earth fluorescence complex comprises europium (Eu), samarium (Sm), dysprosium (Dy), the lanthanide chelate of terbium (Tb) or their potpourri.
The target checking matter of above-mentioned lasting fluorescent-substance markers 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 present invention carries out single to the target checking matter of above-mentioned lasting fluorescent-substance markers or mixes quantitatively or qualitative detection.
The preparation method of lasting fluorescent-substance markers strip according to the invention 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 immersion in the sample pad confining liquid (pH=7.20.03mol/L phosphate buffer+5%BSA ,+0.1%Tween 20), 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 (promptly continuing fluorescent-substance markers thing solution) that continues fluorescence 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 with each assembly of the strip for preparing by sample pad 19, glass fibre membrane pad 20, analyzing film 21, strong adsorptive pads 22, strip reaction end indicating label 23, strip debate know label 24 in order each other 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 subsequent use under drying condition.This shell can be plastics.
The course of work of detection by quantitative according to the invention system is: the to be measured lasting fluorescent-substance markers strip 18 that will be added with sample is put in the strip groove of strip frame 28.The light that light source 1 sends is divided into through fibre bundle 2 and has a determining deviation and the identical two laser of intensity; This two laser is that two bundle directional lights incide dichronic mirror 4 surfaces by collimated illumination lens 3 collimations; Through dichronic mirror 4 reflection back shine synchronously through preceding mirror group 5 be positioned at before mirror group 5 object plane strips 18 detection with 25 with quality control band 26 on excite strip 18 detections with 25 and the lasting fluorescent-substance markers compound of quality control band 26 send the characteristic wavelength reflected fluorescent light; Detection is collected, is gone out back mirror group 7 after seeing through dichronic mirror 4, optical filter 6 elimination veiling glares and get into the electric signal that converts into to be detected in the fluoroscopic image receivers 8 through preceding mirror group 5 with the 25 characteristic wavelength reflected fluorescent light sent with quality control band 26; Electric signal after the conversion is transferred to A/D converter 10 and converts digital signal into behind signal amplifier 9 amplifying signals; The digital signal that data processing and control system 11 gathered, storage transmission is come is also carried out data processing to it, obtains each checking matter and detects and be with 25 OD value OD
Detect bandWith quality control band 26 OD 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 15 automatically, testing result is transferred to output display device 12, accomplishes whole testing process.
Immunochromatography is a dynamic reaction process, continues the siphon on strip of fluorescent-substance markers particle and oozes the process of moving the generation fluorescence excitation 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 possibly there are differences, the quality of the strip product of its production is maybe not can identical.When these strip products are used for the checking matter concentration determination, will certainly have influence on the accurate mensuration of checking matter concentration.For example; Make that the used cellulose membrane source of strip is different, the membrane filtration hole size is different, strip thickness all might not make reactant on the strip of made, can not ooze with identical siphon on an equal basis and move speed and move ahead, the reactant that has in addition might be trapped in strip detect with 25 with quality control band 26 before strip ooze and move in the way.In addition; Environmental baseline of living in can not be just the same during tester's test sample; For example; Field condition detects and differences such as the temperature that exists during indoor detection, humidity also might have influence on the strip reactant and on strip, can not move speed and move ahead with identical oozing, thereby has influence on the accurate mensuration of checking matter concentration.For addressing the above problem, the present invention adopts strip with joining the IC-card 15 corresponding checking matter standard working curves of storage and by IC-card 15 strip quality control band optical density reference monitor value prompting reaction success or failure way being provided simultaneously.Typical curve on the IC-card 15 and strip quality control band optical density reference monitor value can be adjusted with the strip differences between batches, and operation is not only very easy, and have improved the accuracy and the dirigibility that detect.Detection can obtain accurate testing result through looking into standard working curve on the IC-card 15; Simultaneously, in conjunction with the strip quality control band optical density reference monitor value of storage on the IC-card 15, can know whether the strip course of reaction is set up, thereby whether can judge detected result authentic and valid.In addition; The present invention also finds through a large amount of strip reaction tests: strip added example reaction after 3 minutes; Strip reaction end indicating label 23 on the strip can develop the color; The prompting strip is adding a certain particular moment of example reaction after 3 minutes, the strip reactant fully ooze move past detect be with 25 with quality control band 26 positions.Therefore, the present invention is employed in 3 minutes later particular moments of said detection by quantitative system boot just formally makes said system begin sample detection, and these relatively more suitable said system IC-card 15 standard working curves are worked with strip quality control band optical density reference monitor value.The present invention fits to System self-test sweep time with this strip reaction time, and this time parameter also can be identified on the IC-card 15 that matches with strip.
The detection method of application of detection by quantitative according to the invention system realizes through following step:
1. the energized of starting shooting;
2. insert the IC-card 15 that storage has checking matter examination criteria working curve and strip quality control band optical density reference monitor value;
The lasting fluorescent-substance markers strip 18 that 3.) will be added with testing sample is put in the strip groove of strip frame 28;
4. keyboard 14 reads in detected parameters; The checking matter examination criteria working curve that comprises on the IC-card 15 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 are imitated phase, tester's name, detection date, IC-card password etc., and system begins the self check timing automatically afterwards;
5. System self-test finishes; Data processing and control system 11 read IC-card 15 parameters; And drive 28 motions of strip frame and with the light that is fit to illuminator and sends strip 18 is detected and be with 25 to carry out autoscan with quality control band 26; The light signal that scanning obtains converts digital signal into after photoelectricity and mould/number conversion, be transferred to data processing and control system 11;
6. the OD value OD of checking matter characteristic wave strong point in data processing and the control system 11 automatic discriminating digit signals
Detect bandWith quality control band OD 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 15 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 15 provides simultaneously
Quality control band) can judge whether gained checking matter concentration is effective;
7. analysis result is transferred to output display device 12, the output result.
The technical matters that the present invention will solve:
First problem that the present invention will solve provides a kind of based on the test strip quantitative detection system that continues fluorescent-substance markers.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.Said detection system has detection sensitivity height, testing result objective and fair, uses advantages such as flexible.
Second problem that the present invention will solve provides the detection method of application of said detection by quantitative system.
Beneficial effect of the present invention:
(1) can realize quick and precisely that sample is single or polycomponent quantitative or qualitative detection.Lasting fluorescence emission characteristic wavelength fluorescent brightness according to the invention is high, the peak is narrow, symmetrical, the Stokes displacement is big, the life-span is long; With its mark test bar target checking matter correlation molecule; Continue fluorescence intensity through detecting its characteristic wavelength that sends a suitable time delay; Can when effectively eliminate the interference of short life background fluorescence, reach and carry out quantitatively or qualitative detection sending the related objective checking matter molecule that continues fluorescence signal on the strip.Wherein, quantum dot is made to continue fluorescent marker and is had more good characteristic, and the quantum dot of different-grain diameter, heterogeneity and different structure can not only produce the different characteristic wavelength and continue the fluorescence peak spectrum, and the lasting fluorescence peak spectrum that the quantum dot potpourri produces is not overlapping.The quantum dot of selecting different-grain diameter, composition and structure for use is mark test bar target checking matter correlation molecule potpourri and carry out the specific fluorescent signal measuring respectively, can quick and precisely detect sample one-component and polycomponent target checking matter concentration.
(2) can carry out dynamic monitoring to course of reaction.The present invention 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 each target checking matter respective standard working curve cause data processing of joining storage on the IC-card 15 and control system 11 and analyze definite target checking matter concentration automatically.If the detection reaction failure, resulting quality control band OD value OD in the testing process
Quality control bandUnderstand quality control band optical density reference monitor value OD with IC-card 15 storages
Quality control bandThere is very big statistical errors, prompting strip reaction failure.Typical curve on the IC-card 15 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 possibly cause; The present invention 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 in the present invention and also is identified on the IC-card 15 sweep time; But this time parameter order set just began formally sample to be detected after this time; Having guaranteed that the strip reactant fully oozes before system formally begins to detect moves to strip and detects with the luminous energy that is fit to illuminator and sends strip to be detected with quality control band 26 positions with 25 and be with 25 to carry out autoscan with quality control band 26, has improved the accuracy that detects.
(3) can realize the online detection of sample.Fluoroscopic image receiver 8 according to the invention can be CCD, CMOS, photomultiplier, photodiode or phototriode.CCD and CMOS make fluoroscopic image receiver 8, and the test strip quantitative detection system that is constituted is not only applicable to indoor detection, but also can realize the online detection of sample, and vast outdoor location all can conveniently use.The CMOS volume is little, and cost price is cheap.
(4) can make things convenient for tester user that testing result information is carried out flexible real time access and taken outward.IC-card 15 according to the invention links to each other with control system 11 with data processing through the USB serial ports; IC-card 15 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 15 according to own needs in real time, 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 the tester will be added with after the lasting fluorescent-substance markers strip 18 of sample inserts said detection by quantitative system, a few minutes just can accurately obtain accurate testing result.Testing result is objective, and is highly sensitive.
Description of drawings
Fig. 1: based on the test strip quantitative detection system structured flowchart that continues fluorescent-substance markers.
Fig. 2: strip side-looking structural drawing.
Fig. 3: strip plan structure figure.
Fig. 4: HBsAg (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.
Fig. 8: CEA examination criteria working curve.
Fig. 9: Eu
3+The CEA fluorescent spectrum curve of nanoparticle mark.
Sequence number is represented as follows: 1, light source; 2, fibre bundle; 3, collimated illumination lens; 4, dichronic mirror; 5, preceding mirror group; 6, optical filter; 7, back mirror group; 8, fluoroscopic image receiver; 9, signal amplifier; 10, A/D converter; 11, data processing and control system; 12, output display device (like host computer, alphanumeric LCD, LED, audio unit etc.); 13, printer; 14, keyboard; 15, IC-card; 16, driver module; 17, stepper motor; 18, strip; 19, sample pad; 20, glass fibre membrane pad; 21, analyzing film; 22, strong adsorptive pads; 23, strip reaction end indicating label; 24, strip identification label; 25, detect band; 26, quality control band; 27, well (its down corresponding sample pad 19); 28, strip frame; 29, fibre bundle parcel two-beam is fine
Embodiment
(1) continues fluorescent-substance markers test strip quantitative detection system structure
Fig. 1 is the structured flowchart of lasting fluorescent-substance markers test strip quantitative detection system according to the invention.Said detection by quantitative system comprises and continues fluorescent-substance markers strip 18, the strip frame 28 that is used to place strip, illuminator, imaging system, fluoroscopic image receiver 8, signal amplifier 9, A/D converter 10, data processing and control system 11, output display device 12, printer 13, keyboard 14 and one and the supporting IC-card 15 of strip.
The strip frame 28 of said system has the strip groove.The strip groove is used to place lasting fluorescent-substance markers strip 18 to be measured.Said strip 18 is provided with sample pad 19 that mutual overlap joint pastes in order, be coated with the glass fibre membrane pad 20 that continues the fluorescent-substance markers thing, have and detect with 25 and the analyzing film 21 of quality control band 26, strong adsorptive pads 22, strip reaction end indicating label 23 and strip identification label 24.
Said illuminator comprises an excitation source 1, on the output light path of this excitation source 1, is followed successively by fibre bundle 2, collimated illumination lens 3, dichronic mirror 4, preceding mirror group 5, until lasting fluorescent-substance markers strip 18.Excitation source 1 comprises LED or laser diode.The light that fibre bundle 2 sends light source is divided into and has a determining deviation and the identical two laser of intensity, this two laser by collimated illumination lens 3 collimations be incide behind the two bundle directional lights dichronic mirror 4 surfaces through preceding mirror group 5 shine synchronously strip detect with 25 with quality control band 26 on excite strip to detect to send the characteristic wavelength reflected fluorescent light with the lasting fluorescent-substance markers compound of quality control band 26 with 25.
Said imaging system comprises preceding mirror group 5, dichronic mirror 4, optical filter 6, the back mirror group 7 of common optical axis.Dichronic mirror 4 reflectings surface become miter angle with optical axis included angle.Preceding mirror group 5 all adopts isolating construction with back mirror group 7.Illuminator light path part before dichronic mirror 4 is perpendicular with the optical axis of said imaging system, and part after dichronic mirror 4 and said imaging system common optical axis.
Said fluoroscopic image receiver 8 is positioned on mirror group 7 image planes of back.The signal output part of said fluoroscopic image receiver 8 links to each other with said A/D converter 10 through signal amplifier 9.Said fluoroscopic image receiver 8 is collected strips and is detected the characteristic wavelength fluorescence signal that reflects with quality control band 26 with 25, converts thereof into to carrying out signal by signal amplifier 9 behind the electric signal and amplifies.The fluoroscopic image receiver 8 of lasting fluorescent-substance markers test strip quantitative detection system according to the invention has multiple available object, comprises CCD, CMOS, photomultiplier, photodiode, phototriode.Select for use CCD and CMOS to make the lasting fluorescent-substance markers test strip quantitative detection system that fluoroscopic image receiver 8 constituted and both can be used for detecting in the sample chamber, also can be used for the sample real-time online and detect with on-the-spot detection of convenient outdoor sample and use.The CMOS fluoroscopic image receiver product of commercial model has obtained CCD similar image quality now, and volume is little, and power consumption and price are lower than CCD, and has been integrated with that signal amplifies and A/D converter, can signal amplified and has mould/number conversion function.Therefore, when selecting for use CMOS to constitute the present invention to continue the fluorescent-substance markers test strip quantitative detection system, can use signal amplifier 9 and A/D converter 10 no longer in addition more can reduce said detection by quantitative system production cost.Be that CMOS gathers strip detection band and quality control band reflects and transmits the lasting fluorescent-substance markers compound characteristic wavelength fluorescence signal that comes and carry out photoelectricity and mould/number conversion, the digital signal after the conversion is stored, is handled by data processing and control system 11.
Signal amplifier 9 signal output parts behind the signal input part of described A/D converter 10 and the fluoroscopic image receiver 8 link to each other, and the signal output part of A/D converter 10 links to each other with the signal input part of data processing with control system 11.Described A/D converter 10 is used for changing the amplification signal that signal amplifier 9 transmission come into digital signal.
Said data processing and control system 11 are gathered, the next digital signal of storage A/D converter 10 transmission, and digital signal is carried out data processing.Said data processing and control system 11 also are furnished with output display device 12, printer 13, keyboard 14 and an IC-card 15 that matches with strip, and they link to each other with control system 11 with data processing respectively.Also has a Drive and Control Circuit 16 between said data processing and control system 11 and the strip frame 28; 17; This circuit 16, thus 17 said detection by quantitative system is read through its data processing and control system 11 send instruction on the IC-card 15 after the parameter and control 28 motions of strip frame automatically and with the luminous energy that is fit to illuminator and sends strip is detected and be with 25 to carry out autoscan with quality control band 26.Said data processing and control system 11 can corresponding data be handled and single-chip microcomputer, CPU or the PC of Control Software in order to have.
The output display device 12 of said detection by quantitative system can be host computer, alphanumeric LCD, LED, audio unit etc.Said system cooperates different output display devices 12, realizes the quantitative or qualitative detection of sample single component or polycomponent.That is: when said system was used for the sample detection by quantitative, output display device 12 can be host computer or alphanumeric LCD; When said system was used for the sample qualitative detection, output display device 12 can be LED or audio unit etc.
Said IC-card 15 is stored has each checking matter respective detection standard working curve and strip quality control band optical density reference monitor value.Adopt the USB serial communication between IC-card 15 and data processing and the control system 11, during sample detection, IC-card 15 can plug in real time.Each checking matter respective detection standard working curve that IC-card 15 is stored can adopt one of following two kinds of curve modes: the ratio (OD of one of which detection band OD value/quality control band OD value that to be checking matter standard items series concentration record through detection by quantitative according to the invention system with the serial concentration of this checking matter standard items
Detect band/ OD
Quality control band) between the corresponding relation curve; Ratio (the OD of detection band OD value/(detect and be with OD value+quality control band OD value) that also can record through detection by quantitative according to the invention 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 OD 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 15 with standard working curve.In the sample actual detected, if the reaction failure, the OD that sample detection obtains
Quality control bandThe corresponding OD of storage on value and the IC-card 15
Quality control bandValue will produce very big statistical errors.IC-card 15 stores parameter and each batch strip product matches.During sample detection; The operator inserts IC-card 15; Data processing and control system 11 read send after IC-card 15 parameters instruction automatically 28 motions of control strip frame luminous energy that illuminator sends and reflect through dichronic mirror 4 is detected strip be with 25 to carry out autoscan with quality control band 26; Thereby obtain detecting the characteristic frequency fluorescence OD value that sends with quality control band 26 with 25, and can realize that according to the corresponding checking matter standard working curve (the biological respinse success or failure of being pointed out with reference to strip quality control band optical density reference monitor value simultaneously) of IC-card 15 storages checking matter concentration accurately measures.IC-card 15 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 said IC-card 15 storages is: (1) preparation standard items series concentration; (2) place detection by quantitative according to the invention system to record corresponding OD each standard items series concentration
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 the X axle with each standard items series concentration, test result is with OD
Detect band/ OD
Quality control bandRatio is made the Y axle, the drawing standard working curve.Or: make the X axle with each standard items series concentration, test result is with OD
Detect band/ (OD
Detect band+ OD
Quality control band) ratio makes the Y axle, 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 15.
The present invention adopts strip with joining the IC-card 15 corresponding checking matter standard working curves of storage and by IC-card 15 strip quality control band optical density reference monitor value prompting reaction success or failure way being provided.Typical curve on the IC-card 15 and quality control band optical density reference monitor value can be adjusted with the strip differences between batches.The present invention is employed in 3 minutes later particular moments of said detection by quantitative system boot just formally makes said system begin to carry out sample detection, works with the examination criteria working curve and the strip quality control band optical density reference monitor value that are fit to IC-card.This strip reaction time is fitted to System self-test sweep time in the present invention, and its parameter still is identified on the IC-card 15 that matches with strip.
Lasting fluorescence according to the invention means the sustainable life-span material of longer characteristic wavelength fluorescence relatively that sends after one type of certain wavelength excitation of quilt, and it is particularly including quantum dot and nano rare earth fluorescence complex.Wherein, said quantum dot comprises: 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)
2, any one or any several kinds of nano particles among the CdS/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, ZnS:Tb combination, and be that nuclear, silicon dioxide are the core-shell type nano-complex particle of shell by above-mentioned any one quantum dot.Said nano rare earth fluorescence complex comprises europium (Eu), samarium (Sm), dysprosium (Dy), the lanthanide chelate of terbium (Tb) or their potpourri.
The target checking matter of above-mentioned lasting fluorescent-substance markers 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 present invention carries out single to the target checking matter of above-mentioned lasting fluorescent-substance markers or mixes quantitatively or qualitative detection.
Compared with prior art, characteristics of the present invention are: can realize the accurate detection by quantitative of sample one-component or polycomponent, also can carry out sample sxemiquantitative or qualitative detection as required; Utilize fluoroscopic image receiver (like CCD, CMOS), analog to digital converter and data processing and control system (comprising CPU, single-chip microcomputer, PC) and combine the strip reaction time (i.e. the said 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 (like IC-card) can organically combine automatically and continue the fluorescent-substance markers strip and detect and be with 25 to have or not with strong and weak accurately quantitative objective checking matter concentration of quality control band 26 fluorescence or qualitative detection target checking matter; 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, and is easy to operate, do not need other any instrument and equipment and reagent; Just can accurately obtain testing result, high efficiency in a few minutes.
(2) the lasting fluorescent-substance markers test strip quantitative detection system course of work according to the invention
The course of work of detection by quantitative according to the invention system is: the to be measured lasting fluorescent-substance markers strip 18 that will be added with sample is put in the strip groove of strip frame 28.The light that light source 1 sends is divided into through fibre bundle 2 and has a determining deviation and the identical two laser of intensity; This two laser is that two bundle directional lights incide dichronic mirror 4 surfaces by collimated illumination lens 3 collimations; Through dichronic mirror 4 reflection back shine synchronously through preceding mirror group 5 be positioned at before mirror group 5 object plane strips 18 detection with 25 with quality control band 26 on excite strip 18 detections with 25 and the lasting fluorescent-substance markers compound of quality control band 26 send the characteristic wavelength reflected fluorescent light; Detection is collected, is gone out back mirror group 7 after seeing through dichronic mirror 4, optical filter 6 elimination veiling glares and get into the electric signal that converts into to be detected in the fluoroscopic image receivers 8 through preceding mirror group 5 with the 25 characteristic wavelength reflected fluorescent light sent with quality control band 26; Electric signal after the conversion is transferred to A/D converter 10 and converts digital signal into behind signal amplifier 9 amplifying signals; The digital signal that data processing and control system 11 gathered, storage transmission is come is also carried out data processing to it, obtains each checking matter and detects and be with 25 OD value OD
Detect bandWith quality control band 26 OD 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 15 automatically, and afterwards testing result is transferred to output display device 12, accomplish whole testing process.
(3) the detection method of application of lasting fluorescent-substance markers test strip quantitative detection system according to the invention
The detection method of application of detection by quantitative according to the invention system realizes through following step:
1. the energized of starting shooting;
2. insert the IC-card 15 that storage has checking matter examination criteria working curve and strip quality control band optical density reference monitor value;
The lasting fluorescent-substance markers strip 18 that 3.) will be added with testing sample is put in the strip groove of strip frame 28;
4. keyboard 14 reads in detected parameters; The checking matter examination criteria working curve that comprises on the IC-card 15 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 are imitated phase, tester's name, detection date, IC-card password etc., and system begins the self check timing automatically afterwards;
5. System self-test finishes; Data processing and control system 11 read IC-card 15 parameters; And drive 28 motions of strip frame and with the light that is fit to illuminator and sends strip 18 is detected and be with 25 to carry out autoscan with quality control band 26; The light signal that scanning obtains converts digital signal into after photoelectricity and mould/number conversion, be transferred to data processing and control system 11;
6. the OD value OD of checking matter characteristic wave strong point in data processing and the control system 11 automatic discriminating digit signals
Detect bandWith quality control band OD 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 15 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 15 provides simultaneously
Quality control band) can judge whether gained checking matter concentration is effective.
7. analysis result is transferred to output display device 12, the output result.
(1) quantum dot-labeled hepatitis B surface antigen HBsAg strip is made
The quanta dot mark test bar of said detection usefulness is provided with sample pad 19, the glass fibre membrane pad 20 that is coated with quantum dot-labeled HBsAg monoclonal antibody that mutual overlap joint pastes in order, have and detect with 25 and the analyzing film 21 of quality control band 26, strong adsorptive pads 22, strip reaction end indicating label 23 and " HBsAg " strip identification label 24.Detection is with 25 to be coated with the HBsAg monoclonal antibody.Quality control band 26 is coated with two anti-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 assembly preparation of strip
(1) sample pad: select cellulose membrane for use, be cut into 297x15mm 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 297x10mm film piece, put in the elongate slots, add preprepared quantum dot-HBsAg monoclonal antibody bond solution on it, take out the film piece, 37 ℃ of oven dry, fully drying for standby.
(3) analyzing film: select nitrocellulose filter for use; Be cut into 297x25mm film piece; Put in the elongate slots; From film piece base by down from last separated by a distance with some film appearance respectively specking 0.5-5mg/mlHBsAg monoclonal antibody make to detect band 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 and make 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 297x30mm 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 297x5mm, fully drying for standby.
(6) strip is debated the knowledge label: select for use the label paper that is printed with " HBsAg " sign to be cut into the film piece with 297x5mm, fully drying for standby.
2, strip assembling
Each assembly of the strip for preparing is debated the mutual in order overlap joint of knowledge label 24 by sample pad 19, glass fibre membrane pad 20, analyzing film 21, strong adsorptive pads 22, strip reaction end indicating label 23, 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 subsequent use 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 following:
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 OD value (OD respectively
Detect band) and quality control band OD value (OD
Quality control band), calculating mean value and OD
Detect band/ OD
Quality control bandRatio.Make the X axle with HBsAg standard items series concentration, the OD that tries to achieve with each HBsAg standard items series concentration correspondence
Detect band/ OD
Quality control bandRatio is made Y axle drawing standard working curve, and the result sees Fig. 4.
3, write HBsAg examination criteria working curve software, with this Curve Software together with OD
The quality control band value(in the sample actual detected, this OD
The conduct of quality control band valueStrip quality control band optical density reference monitor value) deposits 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 the pH=7.20.03mol/LPB damping fluid before detecting.
2, sample detection:
(1) system power supply is connected in start;
(2) insert the IC-card 15 that storage has HBsAg examination criteria working curve and strip quality control band optical density reference monitor value;
The quanta dot mark test bar 18 that (3) will be added with hepatitis B patient test serum sample is put in the strip groove of strip frame 28;
(4) keyboard 14 reads in detected parameters: comprise HBsAg standard working curve and quality control band optical density reference monitor value, inspection article title, inspection article numbering, strip lot number, strip effect phase, detection date, tester's name, the IC-card password of IC-card 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 and control system 11 read IC-card 15 parameters; And drive 28 motions of strip frame and with the light that is fit to illuminator and sends strip 18 is detected and be with 25 to carry out autoscan with quality control band 26; The light signal that scanning obtains converts digital signal into after photoelectricity and mould/number conversion, be transferred to data processing and control system 11;
(6) detect the OD value (OD that is with the characteristic wave strong point in data processing and the control system 11 automatic discriminating digit signals
Detect band) and quality control band OD value (OD
Quality control band), calculate OD automatically
Detect band/ OD
Quality control bandRatio, and calculate the HBsAg concentration in the blood serum sample automatically according to the HBsAg standard working curve that stores in the IC-card.Detecting its HBsAg concentration like the present embodiment blood serum sample through said detection by quantitative system is 205.9563pg/ml;
(7) output display device 12 shows testing result;
(8) system print test results report.
3, hepatitis B patient blood serum sample HBsAg ultimate density is calculated:
HBsAg concentration * serum diluting multiple that HBsAg ultimate density (pg/ml)=system records in the blood serum sample
In the present embodiment, said blood serum sample HBsAg ultimate density (pg/ml)=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.
(1) the quantum dot-labeled multinomial strip of tumor markers one inspection is made
Said strip 18 is provided with sample pad 19, glass fibre membrane pad 20 that mutual overlap joint pastes in order, have and detect with 25 and the analyzing film 21 of quality control band 26, strong adsorptive pads 22, strip reaction end indicating label 23 and " AFP/CEA/PSA " strip identification label 24.Glass fibre membrane pad 20 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 with 25 be coated with AFP antibody, CEA antibody and PSA antibody potpourri; Quality control band 26 is coated with two anti-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 assembly preparation of strip
(1) sample pad: select cellulose membrane for use, be cut into 297x15mm 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 297x10mm 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) and on it, take out the film piece; 37 ℃ of oven dry, fully drying for standby.
(3) analyzing film: select nitrocellulose filter for use; Be cut into 297x25mm film piece; Put in the elongate slots; Be with by following the detection from film piece base from last potpourri work with some film appearance 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 some film appearance 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 297x30mm 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 297x5mm, fully drying for standby.
(6) strip is debated the knowledge label: select for use the label paper that is printed with " AFP/CEA/PSA " sign to be cut into the film piece with 297x5mm, fully drying for standby.
2, strip assembling
Each assembly of the strip for preparing is debated the mutual in order overlap joint of knowledge label 24 by sample pad 19, glass fibre membrane pad 20, analyzing film 21, strong adsorptive pads 22, strip reaction end indicating label 23, 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 subsequent use 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 following:
1, 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, 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 OD value (OD respectively
Detect band) and quality control band OD value (OD
Quality control band), obtain mean value and OD
Detect band/ OD
Quality control bandRatio.Make the X axle 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 axle drawing standard working curve, and the result sees Fig. 6.
3, write tumor markers examination criteria working curve software, with this Curve Software together with OD
The quality control band value(in the sample actual detected, this OD
The conduct of quality control band valueStrip quality control band optical density reference monitor value) deposits 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=7.20.03mol/L PB damping fluid before detecting.
2, sample detection:
(1) system power supply is connected in start;
(2) insert the IC-card 15 that storage has AFP, CEA, PSA examination criteria working curve and strip quality control band optical density reference monitor value;
The quanta dot mark test bar 18 that (3) will be added with tumor patient test serum sample is put in the strip groove of strip frame 28;
(4) keyboard 14 reads in detected parameters: AFP, CEA, PSA standard working curve and the quality control band optical density reference monitor value, inspection article title, inspection article 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 and control system 11 read IC-card 15 parameters; And drive 28 motions of strip frame and with the light that is fit to illuminator and sends strip 18 is detected and be with 25 to carry out autoscan with quality control band 26; The light signal that scanning obtains converts digital signal into after photoelectricity and mould/number conversion, be transferred to data processing and control system 11;
(6) detect the OD value (OD that is with the characteristic wave strong point in data processing and the control system 11 automatic discriminating digit signals
Detect band) and quality control band OD value (OD
Quality control band), calculate OD automatically
Detect band/ OD
Quality control bandRatio, and calculate AFP, CEA and PSA concentration in the blood serum sample automatically according to the AFP that stores in the IC-card, CEA, PSA standard working curve.Is that 1753.4526pg/ml, CEA concentration are that 878.3892pg/ml, PSA concentration are 983.4257pg/ml like the present embodiment blood serum sample through said detection by quantitative its AFP of detection of system;
(7) output display device 12 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:
Blood serum sample AFP ultimate density (pg/ml)=1753.4526pg/ml * 10=17534.526pg/ml
Blood serum sample CEA ultimate density (pg/ml)=878.3892pg/ml * 10=8783.892pg/ml
Blood serum sample PSA ultimate density (pg/ml)=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.
Embodiment 4 nano rare earth fluorescence complexs are made to continue the fluorescent-substance markers strip and are detected S-CEA CEA (individual event detection)
The nano rare earth fluorescence complex is with Eu
3+Nanoparticle is an example.
(1) strip is made
Eu
3+Nanoparticle mark test bar 18 is provided with sample pad 19, glass fibre membrane pad 20 that mutual overlap joint pastes in order, have and detect with 25 and the analyzing film 21 of quality control band 26, strong adsorptive pads 22, strip reaction end indicating label 23 and " CEA " strip identification label 24.Glass fibre membrane pad 20 is coated with Eu
3+The CEA monoclonal antibody of nanoparticle mark; Detection is with 25 to be coated with CEA antibody; Quality control band 26 is coated with two anti-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 assembly preparation of strip
(1) sample pad: select cellulose membrane for use, be cut into 297x15mm 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 297x10mm film piece, put in the elongate slots, add preprepared Eu
3+The CEA monoclonal antibody solution of nanoparticle mark takes out the film piece on it, 37 ℃ of oven dry, fully drying for standby.
(3) analyzing film: select nitrocellulose filter for use; Be cut into 297x25mm film piece; Put in the elongate slots, by down separated by a distancely making to detect band, make quality control band with a film appearance 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 with some film appearance specking 0.5-5mg/ml CEA antibody from last from film piece base; 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 297x30mm 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 297x5mm, fully drying for standby.
(6) strip is debated the knowledge label: select for use the label paper that is printed with " CEA " sign to be cut into the film piece with 297x5mm, fully drying for standby.
2, strip assembling
Each assembly of the strip for preparing is debated the mutual in order overlap joint of knowledge label 24 by sample pad 19, glass fibre membrane pad 20, analyzing film 21, strong adsorptive pads 22, strip reaction end indicating label 23, 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 subsequent use under drying condition.This shell can be plastics.
(2) standard working curve is drawn and storage
Fig. 8 is a CEA examination criteria working curve.Its method for making is following:
1, 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, and the CEA standard items are made into 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 CEA examination criteria working curve
Every part of CEA standard items series concentration is used 10 Eu respectively
3+The CEA strip of nanoparticle mark is provided with under the condition in identical systems and detects 10 times, reads to such an extent that it detects band OD value (OD respectively
Detect band) and quality control band OD value (OD
Quality control band), obtain mean value and OD
Detect band/ (OD
Quality control band+ OD
Quality control band) ratio.Make the X axle with each CEA standard items series concentration, the OD that tries to achieve with each CEA standard items series concentration correspondence
Detect band/ (OD
Quality control band+ OD
Quality control band) ratio makes Y axle drawing standard working curve, the result sees Fig. 8.
3, write CEA examination criteria working curve software, with this Curve Software together with OD
The quality control band value(in the sample actual detected, this OD
The conduct of quality control band valueStrip quality control band optical density reference monitor value) deposits IC-card in.
(3) blood serum sample CEA detects
1, sample source: blood serum sample is provided by certain healthcare hospital for women & children laboratory.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 15 that storage has CEA examination criteria working curve and strip quality control band optical density reference monitor value;
(3) will be added with the Eu of test serum sample
3+The CEA strip 18 of nanoparticle mark is put in the strip groove of strip frame 28;
(4) keyboard 14 reads in detected parameters: comprise CEA standard working curve and quality control band optical density reference monitor value, inspection article title, inspection article numbering, strip lot number, strip effect phase, detection date, tester's name, the IC-card password of IC-card etc., System self-test is made as 5 minutes (containing delay 400 μ S) sweep time, system begins the self check timing automatically afterwards;
(5) System self-test finishes; Data processing and control system 11 read IC-card 15 parameters; And drive 28 motions of strip frame and with the light that is fit to illuminator and sends strip 18 is detected and be with 25 to carry out autoscan with quality control band 26; The light signal that scanning obtains converts digital signal into after photoelectricity and mould/number conversion, be transferred to data processing and control system 11;
(6) detect the OD value (OD that is with the characteristic wave strong point in data processing and the control system 11 automatic discriminating digit signals
Detect band) and quality control band OD value (OD
Quality control band), calculate OD automatically
Detect band/ (OD
Quality control band+ OD
Quality control band) ratio, and calculate the CEA concentration in the blood serum sample automatically according to the CEA standard working curve that stores in the IC-card.Detecting its CEA concentration like the present embodiment blood serum sample through said detection by quantitative system is 586.4602pg/ml;
(7) output display device 12 shows testing result;
(8) system print test results report.
3, blood serum sample CEA ultimate density is calculated:
CEA concentration * serum diluting multiple that CEA ultimate density (pg/ml)=system records in the blood serum sample
In the present embodiment, said blood serum sample CEA ultimate density (pg/ml)=586.4602pg/ml * 10
＝5864.602pg/ml
Fig. 9 is Eu
3+The CEA fluorescent spectrum curve of nanoparticle mark.Wherein, I is for detecting the CEA spectrum peak in the band, and II is the Quality Control thing spectrum peak in the quality control band.
Top embodiment and accompanying drawing thereof only are in order to further specify the present invention, and those skilled in the art should not limit protection scope of the present invention with this.Special needs to be pointed out is; The lasting fluorescence that detection by quantitative according to the invention system is fit to is except that quantum dot, nano rare earth fluorescence complex; Can also be that some other stimulated luminescence excites the lasting fluorescence that can produce relative long life characteristics wavelength fluorescent, the organic polymer fluorescent nano particles that for example aggregates into etc. by organic monomer styrene, acrylic acid.Therefore, every to the lasting fluorescence implication of indication of the present invention and any other technical scheme that is equal to replacement or equivalent transformation formation of said lasting fluorescent-substance markers test strip quantitative detection system employing, all drop in the protection domain of claim of the present invention.
Claims (13)
1. one kind based on the test strip quantitative detection system that continues fluorescent-substance markers, it is characterized in that: it comprises and continues fluorescent-substance markers strip (18), the strip frame (28) that is used to place strip, illuminator, imaging system, fluoroscopic image receiver (8), signal amplifier (9), A/D converter (10), data processing and control system (11), output display device (12), printer (13), keyboard (14) and one and the supporting IC-card (15) of strip:
Said illuminator comprises an excitation source (1), on the output light path of this excitation source (1), is followed successively by fibre bundle (2), collimated illumination lens (3), dichronic mirror (4), preceding mirror group (5), until lasting fluorescent-substance markers strip (18);
Said imaging system comprises preceding mirror group (5), dichronic mirror (4), optical filter (6), the back mirror group (7) of common optical axis; Dichronic mirror (4) reflecting surface becomes miter angle with optical axis included angle; Preceding mirror group (5) and back mirror group (7) all adopt isolating construction; Part is perpendicular with the optical axis of said imaging system before at dichronic mirror (4) for the illuminator light path, and the illuminator light path is at dichronic mirror (4) part afterwards and said imaging system common optical axis;
Said fluoroscopic image receiver (8) is positioned on mirror group (7) image planes of back; The signal output part of fluoroscopic image receiver (8) links to each other with A/D converter (10) through signal amplifier (9), and said A/D converter (10) links to each other with control system (11) with said data processing; Said output display device (12), printer (13), keyboard (14) and IC-card (15) link to each other with control system (11) with said data processing respectively;
Between said data processing and control system (11) and the strip frame (28) Drive and Control Circuit (16 is arranged; 17); Said data processing and control system (11) read IC-card (15) parameter and send instruction and control automatically through this Drive and Control Circuit (16,17) that strip frame (28) motion detects band (25) with the light that is fit to illuminator and sends to strip and quality control band (26) carries out autoscan;
The light that light source (1) sends is divided into through fibre bundle (2) and has a determining deviation and the identical two laser of intensity; This two laser is that two bundle directional lights incide dichronic mirror (4) surface by collimated illumination lens (3) collimation; Shine synchronously through preceding mirror group (5) through dichronic mirror (4) reflection back and to excite strip (18) detection to be with the lasting fluorescent-substance markers compound of (25) and quality control band (26) to send the characteristic wavelength reflected fluorescent light on detection band (25) and the quality control band (26) of mirror group (5) object plane strip (18) before being positioned at; Detecting band (25) and quality control band (26) reflected fluorescent light mirror group (5) before same collects, gets into the electric signal that converts into to be detected in the fluoroscopic image receiver (8) through going out back mirror group (7) behind dichronic mirror (4), optical filter (6) the elimination veiling glare; Electric signal after the conversion is transferred to A/D converter (10) and converts digital signal into and be transferred to data processing and control system (11) is stored and data processing behind signal amplifier (9) amplifying signal; Data processing and control system (11) be automatically to detecting that band (25) the characteristic frequency fluorescence optical density that transmission comes with quality control band (26) is discerned and carrying out concentration according to the checking matter standard working curve of IC-card (15) storage and calculate and technical Analysis, with result transmission to output display device (12).
2. lasting fluorescent-substance markers test strip quantitative detection system according to claim 1 is characterized in that: said excitation source (1) comprises LED or laser diode.
3. lasting fluorescent-substance markers test strip quantitative detection system according to claim 1 is characterized in that: said fluoroscopic image receiver (8) can be CCD, CMOS, photomultiplier, photodiode or phototriode.
4. lasting fluorescent-substance markers test strip quantitative detection system according to claim 1 is characterized in that: said data processing and control system (11) can corresponding data be handled and single-chip microcomputer, CPU or the PC of Control Software in order to have.
5. lasting fluorescent-substance markers test strip quantitative detection system according to claim 1 is characterized in that: said output display device (12) can be host computer, alphanumeric LCD, LED or audio unit.
6. lasting fluorescent-substance markers test strip quantitative detection system according to claim 1; It is characterized in that: said lasting fluorescent-substance markers strip (18) is the lasting fluorescent-substance markers strip (18) that said detection by quantitative system matches, and is provided with sample pad (19), glass fibre membrane pad (20) that mutual overlap joint pastes on it in order, has the analyzing film (21) that detects band (25) and quality control band (26), adsorptive pads (22), strip reaction end indicating label (23) and strip identification label (24) by force.
7. lasting fluorescent-substance markers test strip quantitative detection system according to claim 6; It is characterized in that: the glass fibre membrane pad (20) of said lasting fluorescent-substance markers strip is coated with the associated biomolecule molecule that the target checking matter detects that is used for that continues fluorescent-substance markers; Detect band (25) and be coated with and be used for the relevant special biomolecule that the target checking matter detects, quality control band (26) is coated with and comprises the two Quality Control things that resist.
8. lasting fluorescent-substance markers test strip quantitative detection system according to claim 7 is characterized in that: the said lasting fluorescence of the glass fibre membrane pad (20) of wherein said strip is the sustainable material that sends relative longer characteristic wavelength fluorescence of life-span after one type of certain wavelength excitation of ability quilt.
9. lasting fluorescent-substance markers test strip quantitative detection system according to claim 8; It is characterized in that: wherein said lasting fluorescence is a quantum dot, 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)
2, any one or any several kinds of nano particles among the CdS/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, ZnS:Tb combination, and be that nuclear, silicon dioxide are the core-shell type nano-complex particle of shell by above-mentioned any one quantum dot.
10. lasting fluorescent-substance markers test strip quantitative detection system according to claim 8 is characterized in that: wherein said lasting fluorescence is the nano rare earth fluorescence complex, comprises the lanthanide chelate of Eu, Sm, Dy, Tb or their potpourri.
11. lasting fluorescent-substance markers test strip quantitative detection system according to claim 1; It is characterized in that: said IC-card (15) is stored has checking matter examination criteria working curve, its: or the ratio OD of the detection band OD value/quality control band OD value that records through system according to the invention for checking matter standard items series concentration and the serial concentration of this checking matter standard items
Detect band/ OD
Quality control bandBetween the corresponding relation curve, or the ratio OD of detection band OD value/(detect and be with OD value+quality control band OD value) that records through system according to the invention for checking matter standard items series concentration and this checking matter standard items series concentration
Detect band/ (OD
Detect band+ OD
Matter The control band) between the corresponding relation curve.
12. lasting fluorescent-substance markers test strip quantitative detection system according to claim 1 is characterized in that: said IC-card (15) is stored has strip quality control band optical density reference monitor value.
13. the detection method of application of a lasting fluorescent-substance markers test strip quantitative detection system as claimed in claim 1 is characterized in that may further comprise the steps:
1. the energized of starting shooting;
2. insert IC-card (15);
The lasting fluorescent-substance markers strip (18) that 3.) will be added with testing sample is put in the strip groove of strip frame (28);
4. keyboard (14) reads in detected parameters; Comprise that checking matter examination criteria working curve that IC-card (15) goes up 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, detection date, system begins the self check timing automatically afterwards;
5. System self-test finishes; Data processing and control system (11) read IC-card (15) parameter; And the light that driving strip frame (28) motion is sent with suitable illuminator carries out autoscan to strip (18) detection band (25) and quality control band (26); The light signal that scanning obtains converts digital signal into after photoelectricity and mould/number conversion, be transferred to data processing and control system (11);
6. the automatic OD value OD of checking matter characteristic wave strong point in the discriminating digit signal of data processing and control system (11)
Detect bandWith quality control band OD 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 calculate checking matter concentration automatically, the quality control band optical density reference monitor value OD that IC-card (15) provides simultaneously according to the corresponding checking matter examination criteria working curve of storage in the IC-card (15)
Quality control bandAssist to judge whether measured checking matter concentration is effective;
7. analysis result is transferred to output display device (12), the output result.
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| CN2009100582980A CN101592659B (en) | 2009-02-09 | 2009-02-09 | System and method for quantitative detection of test strips on basis of continuous fluorescent-substance markers |
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