CN102087215A - Fluorescent quantitative detection instrument - Google Patents

Abstract

The invention provides a fluorescent quantitative detection instrument. The instrument comprises an excitation light source module, a photoelectric conversion module, a control analysis module and a software system, wherein the excitation light source module comprises a first laser module component and a second laser module component; the first laser module component is used for transmitting an excitation beam A to a detection region on a reagent strip; the second laser module component is used for transmitting an excitation beam B to a coding region on the reagent strip; the photoelectric conversion module is used for receiving a reflected fluorescent signal C and a laser signal D and carrying out photoelectric signal conversion; the control analysis module comprises a main circuit, a motor and a display screen, and is used for processing received electric signals output by the photoelectric conversion module; the fluorescent quantitative detection instrument is provided with a reagent strip slot for placing a reagent strip and an ID card slot for placing a scaler chip ID card; and the motor is used for driving the reagent strip to move. The fluorescent quantitative detection instrument has the following beneficial effect: the specially designed optical system and compensation circuit are adopted, so the fluorescent quantitative detection instrument has higher sensitivity and accuracy in immunofluorescent quantitative detection.

Description

The fluorescent quantitation detector

 

[technical field]

The invention belongs to field of medical examination, relate in particular to a kind of fluorescent quantitation detector that is used to catch fluorescence signal.

[background technology]

The immunofluorescence detection technique is to adopt fluorescein to carry out mark first and succeed since Coons equals nineteen forty-one.This technology of carrying out Antigen Location with the fluorescent material labelled antibody is called fluorescent antibody technics (fluorescent antibody technique).Method with fluorescence antibody spike or inspection corresponding antigens claims fluorescence anti-body method; Method with known fluorescent antigen label spike or inspection corresponding antibodies claims the fluorescent antigen method.These two kinds of method general name immunofluorescence techniques, because fluorchrome not only can combine with antibody globulin, be used for detecting or locating various antigens, also can with other protein bound, be used for detecting or location antibody, but the fluorescent antigen technology is seldom used in real work, so people's custom is called fluorescent antibody technics, or is called immunofluorescence technique.More commonly used with the fluorescence antibody method.

Be called immunofluorescence cell (or tissue) chemical technology with the immunofluorescence technique demonstration and the methods such as cell or tissue endoantigen or haptens material of checking.The principal feature of this technology is: high specificity, susceptibility height, speed are fast.Major defect is: the unspecific staining problem solves as yet fully, the objectivity deficiency that the result judges, and technical program also more complicated.

Fluorescent immune method also can further divide and do some kinds by reaction system and quantivative approach difference.Compare with radioimmunology, the "dead" pollution of fluorescent immune method, and easy and simple to handle mostly, be convenient to promote.Traditional medical immunoassay device is a kind of device that is used to detect antigen, antibody response; its principle of work is to utilize the power of the electric signal that antigen, antibody response produce to determine the size of antigen concentration; the signal that is produced by antigen, antibody id reaction detects the concentration of antigen; and the content of antigen, antibody is generally the nanogram level; therefore signal is very faint; only in several millivolts; be a kind of electrophoresis process very slowly to the compound of antigen, antibody together; so recombination time is long; this brings difficulty to testing, therefore makes slow progress.

Because the background in the general fluorometric assay is than problems such as height, immunofluorence technic is used for quantitative measurement certain difficulty.Development in recent years several special fluorescence immunoassay, the same with enzyme immunoassay (EIA) and radiommunoassay, in clinical examination, use.

Fluorescent latex mark chromatography detection technique is to continue after the colloidal gold-labeled method, on the basis of latex agglutination test, grow up, as a kind of immunological method, it is the combination of immune affine technology, engram technology, immunolabelling technique and chromatographic technique, and utilizes fluorescent quantitation detector detection by quantitative fluorescence signal.Owing to but it equally has quick, easy and simple to handle, stable reagent room temperature accumulating, is difficult for pollution characteristics and utilizes the advantage of fluorescence detector detection by quantitative to develop rapidly with golden mark technology.

Fluorescent quantitation detecting instrument hardware components generally is made up of computer, IO interface, analog to digital converter, scan control circuit, photoelectric switching circuit, background compensation circuit, display etc.But the common volume of fluorescent quantitation detector general in the prior art is bigger, is not easy to real-time test, has limited it and has had inconvenient traffic, the development of under-developed area.

And because the combination and the marker material of antigen-antibody are all very expensive, can not be as chemical reagent, make the large tracts of land colour developing, background has impurity such as water, blood, marker material simultaneously, and reason such as inhomogeneous has increased the difficulty and the precision that detect in process of osmosis, how small at the volume that keeps the POCT product, improving the stability and the sensitivity that detect the time easy to carry is present technique field problem demanding prompt solution.

[summary of the invention]

The object of the present invention is to provide the fluorescent quantitation detector that a kind of detection is easier to, precision is high.

For realizing the object of the invention, provide following technical scheme:

The present invention provides a kind of immunofluorescence detection system in order to address the above problem, and designs special optical system, and compensating circuit, and the uneven undesired signal that produces of filtration and infiltration is optimized the problem that solves immunofluorescence quantitative test medium sensitivity and degree of accuracy.

Fluorescent quantitation detector of the present invention, it comprises excitation source module, photoelectric conversion module, control analysis module and software systems, described excitation source module comprises the first laser module assembly and the second laser module assembly; The first laser module assembly is used for the detection zone launching excitation light bundle A to reagent strip, through reflecting fluorescence signal C; The second laser module assembly is used for the code area launching excitation light bundle B on reagent strip, through reflecting laser signal D; Described photoelectric conversion module is used to receive the fluorescence signal C and the laser signal D of reflection and carries out the photosignal conversion; Described control analysis module comprises main circuit, motor, display screen, and the control analysis module is used to handle the electric signal that receives photoelectric conversion module output and handles and outputs to display screen and show; Described fluorescent quantitation detector is provided with reagent strip slot and the ID card slot that is used to place reagent strip and calibration chip id card; This motor is used to drive reagent strip and moves.

Preferably, this fluorescent quantitation detector also comprises a microprinting machine, is used to print testing result.

Preferably, this photoelectric conversion module comprises a signal compensation circuit, reduces the generation of undesired signal.

Preferably, what the described first laser module assembly sent is the laser of 470NM wavelength, and when reagent strip was shone, its fluorescent material can send the light wave of 530nm.

Preferably, described display screen adopts the touching display screen device, is used for instruction input and data presentation.

Preferably, there is the code area on described matching used reagent strip surface, and this code area is read by the laser module assembly and import main circuit after the photosignal conversion, for distinguishing different test items.The item recognition module is arranged in the described analysis module, be used to discern the code area on matching used reagent strip surface, this code area can be for distinguishing different test items.

Preferably, store the calibration curve of respective batch reagent card in the described ID fastening mark chip.

Preferably, in the described control analysis module quality control module is arranged, be used for reading the calibration curve of calibration chip id card, and carry out data according to calibration curve and judge.

Preferably, also comprise a built-in storage unit, can store up-to-date a plurality of testing results.

Preferably, described software systems comprise initialization module, start calibration module, system parameter setting module, bar code acquisition module and test sample analysis module.

Adopt the detection method of fluorescent quantitation detector of the present invention, it adopts aforesaid fluorescent quantitation detector, and it comprises the steps:

A. the reagent strip of reaction injects the reagent strip slot in advance;

B. the excitation source module is sent the detection zone that excitation source is radiated at the test agent bar, and the fluorescent latex on the detection zone is launched fluorescence signal under the effect of exciting light;

C. the fluorescence signal launched of detection zone is caught and finishes the conversion of photosignal by the photoelectric conversion module that is provided with in the fluorescent quantitation detector;

D. through after the control analysis resume module result being shown.

The contrast prior art, the present invention has the following advantages:

Fluorescent quantitation detector of the present invention adopts the optical system and the compensating circuit of particular design, makes the immunofluorescence detection by quantitative sensitive more and accurate.

[description of drawings]

Fig. 1 is the structural representation of fluorescent quantitation detector of the present invention;

Fig. 2 is the structure and the workflow rough schematic of fluorescent quantitation detector of the present invention;

[embodiment]

Describe the present invention in detail below in conjunction with Fig. 1 and Fig. 2 and specific embodiment.

Fluorescent quantitation detector of the present invention, compriseexcitation source module 10, photoelectric conversion module,control analysis module 3, described excitation source module comprises the first laser module assembly 1 and the second laser module assembly, 2 two parts, and this photoelectric conversion module comprises first photoelectric conversion module 41 and second photoelectric conversion module 42; The first laser module assembly 1 is used for thedetection zone 53 launching excitation light bundle A to reagent strip; The second laser module assembly 2 is used for the code area launching excitation light bundle B on reagent strip; Described photoelectric conversion module 41 and 42 is used to receive the fluorescence signal C and the laser signal D of reflection and carries out the photosignal conversion; Describedcontrol analysis module 3 comprisesmain circuit 31,motor 34,display screen 33; Automatically the control analysis module is used to handle the electric signal that receives photoelectric conversion module output and handles and outputs to displayscreen 33 and show, and can pass throughdisplay screen 33 input instructions; Described fluorescent quantitation detector is provided with reagent strip slot and the ID card slot that is used to placereagent strip 5 and calibrationchip id card 32, and calibrationchip id card 32 is used to import test item, lot number and related data; This motor is used to drive about reagent strip scanning and moves.51 is sample application zone among the figure.

This first laser module assembly 1 comprises excitation source 11,spectroscope 12, excitation beam A is transmitted in thedetection zone 53 ofreagent strip 5 throughlens 13, through reflecting fluorescence signal C, shine first photoelectric conversion module 41 through optical filter 14,lens 15, grating 16, importmain circuit 31 then.

This second laser module assembly 2 compriseslight source 21,spectroscope 22 and lens 23, themain circuit 31 controls second laser module assembly 2 launching excitation light bundle B, be transmitted intocode area 52 on thereagent strip 5 throughspectroscope 22 and lens 23, through reflecting fluorescence signal C, through second photoelectric conversion module 42, importmain circuit 31 then again.

First photoelectric conversion module 41 is used to receive the signal of reflected fluorescent light C, amplifies by opto-electronic conversion, signal, and the photon intensity that obtains also is converted to the output of the product of detection concentration on display screen.

Second photoelectric conversion module 42 is used to receive the signal of reflector laser D, andcode area 52 information on the reading reagent bar are imported main circuit with signal and distinguished different test items for system.

The excitation source that this excitation source module is sent is radiated at the detection zone of test agent bar, and the fluorescence signal that detection zone sent is caught and be converted to electric signal by photoelectric conversion module, and the output electric signal is to the control analysis module, and output display screen shows.

Software systems of the present invention by the detection control algolithm program of machine intimate with form based on softwares such as the analysis of PC platform and databases.(whether this detector can be connected with pc, will detect data and export to upward preservation management of pc, also can not be connected with pc, directly is kept in the storage card, connect, and pattern can be selected)

Control analysis module 3 comprisesmain circuit 31,motor 34, coded scanning device and display circuit; The motion of main 31 circuit control detection probe.

In a preferred embodiment of the present invention, assembled a miniature printer, be used to print testing result.

In another preferred embodiment of the present invention, also added a signal compensation circuit in first photoelectric conversion module 41, be used to reduce undesired signal.

During detection, instrument is discerned test item automatically by the code area on the second laser module assembly, the 2 scanning reagent strips, and test item is presented on the LCDs, further confirms test item when beginning to detect for the operator.And in the reagent strip surface design direction arrow, the direction accuracy when guaranteeing that it inserts detector.

Preferred a kind of embodiment, display adopt liquid crystal display to show or screen displaying.

Software systems of the present invention comprise following functional module:

Instrument initialization module: each working cell of system (serial port) initialization is set;

Instrument start calibration module: whether inspection apparatus duty and static parameter be normal;

The instrument system parameter is provided with module: setting and change Instrument working state and parameter (date, time etc.),

Instrument bar code collection analysis module: gather reagent strip code area information, analyze input reagent strip corrected parameter, test sample book parameter, handle corresponding test event parameter.

Instrument test sample analysis module: collecting test fluorescent belt information, according to the parameter of bar code collection analysis, utilize the concentration of the algorithm computation detected material of pre-programmed.

Fluorescent latex labelled antibody/antigen is with the latex microsphere and the protein that contains to be measured antibody/antigen and all kinds of different fluorescein covalent bond of diameter range at 0.01 μ m～1 μ m, the carboxyl that utilizes the latex microsphere macromolecular substances to have, amino, group binding antibodies such as hydroxyl, utilize the fluorescein can emitting fluorescence under the exciting light effect, when the antibody/antigen of this fluorescent latex mark with detect the latex labeled complex that forms after corresponding antigens/antibodies enough in the sample, when the antibody/antigen of fluorescent latex mark is built up in a large number at corresponding part place, utilize fluorescent quantitation detector provided by the invention, fluorescein by detection zone accumulation on the light source activation film, the fluorescence that fluorescein is launched is received by the relevant detection instrument, and pass through light-to-current inversion, processes such as photoelectricity conversion change into electric signal with photosignal, and by the automatic control system that is provided with in the instrument signal is exported, demonstrate final quantitative result.Because the difference of the kind of the fluorescein of selecting, its excitation/emission light wavelength λ and automatic control system also can be different.

Because laser has good unicity, coherence, directivity, the immunofluorescence detector uses laser as excitation source in the present invention, excite incident light by laser tube, incident illumination is mapped to the detection zone in the reagent strip, fluorescent material on C, the T line is excited, produce a kind of new wavelength thereby electronic transition takes place, be greater than generally speaking and excite light wavelength.The emission light that produces is received by photomultiplier and is converted into electric signal, and the power of electric signal is strict relevant with the fluorescent material molecular amounts, and the electric signal that receiver is changed out is translated into accessible numeral with A/D converter after amplifying by amplifying circuit.Because immune response is a dynamic change procedure, so instrument obtains the pulse signal of two varying strengths, by the treated photon pulse number that converses of the difference of two signals by carry out mensuration twice in fixing Preset Time.

In preferred embodiment of the present invention, what fluorescence detecting system adopted is the laser of 470NM wavelength, and when reagent strip was shone, its fluorescent material can send the light wave of 530nm.

The immunofluorescence detection technique is with the fluorescence emulsion particle thing of marking, and chromatography strip is composited by multiple material.The detecting instrument hardware components generally is made up of computer, IO interface, analog to digital converter, scan control circuit, photoelectric switching circuit, background compensation circuit, display etc.In order to preserve test result, is furnished with mini-printer.

In order to improve to improve the reliability and the accuracy of testing result.The present invention works out special software according to the infiltration rule of water, the inhomogeneous undesired signal that causes of compensation infiltration; According to different test-strips, different lot numbers, the control test duration.

Among the present invention, the relation of optical density value and concentration is represented with detection curve, when measuring, can automatically optical density value be converted to concentration value and demonstration at every turn.Difference by the fluorescence signal value in detection zone, Quality Control district on the test agent bar becomes certain ratio with the variable concentrations of analyte, can calculate the concentration of analyte the unknown sample from curve.

The detection method of fluorescent quantitation detector may further comprise the steps:

A. the reagent strip of reaction injects the reagent strip slot in advance;

B. the fluorescent optics system that is provided with in the fluorescent quantitation detector comprises light source, outer light path, monochromator etc.Light source converges light by outer light path, be projected on the entrance slit of monochromator the parasitic light beyond the monochromator filtering analytical line.

The single excitation source that penetrates from monochromator is radiated at the detection zone of test agent bar, and the fluorescent latex on the detection zone is launched fluorescence signal under the effect of exciting light.

C. the fluoroscopic examination module that is provided with in the fluorescent quantitation detector of the fluorescence signal launched of detection zone is caught, and is finished the conversion of photosignal by the fluoroscopic examination module.The light that the fluoroscopic examination module is launched test agent bar detection zone passes through light-to-current inversion, accept incident light, launch photoelectron and make its multiplication, realize the amplification of photoelectron signal, by solid-state detector photoelectron signal is converted to electric signal again, by the electric signal sensing circuit electric signal is exported, after automatic software analysis and Control system handles, the result is shown by digital display screen.

As what detect that HbA1c uses is immune competition law.During whole blood mixing after detecting damping fluid and having added the haemolysis damping fluid, fluorescently-labeled anti-HbA1c antibody combines with HbA1c in the blood sample, then after this sample mixed liquor joins the sample application zone of reagent strip, HbA1c in the sample and the glycosylated hemoglobin that is fixed on the reagent strip then can combine competitively with detection antibody (fluorescent-labeled antibody), behind the molecular balance, HbA1c in the sample is many more, it is just few more to be fixed on the chance that the glycosylated hemoglobin on the reagent strip combines with fluorescent-labeled antibody, reads fluorescence intensity shown in the reagent strip at last.The strong and weak amount with HbA1c of fluorescence signal is inversely proportional to.Fluorescence detecting system detects HbA1c concentration and total hemoglobin concentration.Instrument is that ratio (%) is presented on the screen with these two Parameters Transformation, be exactly the relative concentration of HbA1c (total accounting for the ratio of Hb).

What detect that CRP albumen uses is dual-antigen sandwich method, and during detection, blood sample dilution back adds reagent strip, on CRP antigen in the blood and the reagent strip with the fluorescently-labeled CRP antibody 1 formation antigen antibody complex that combines; Caught by the anti-CRP antibody 2 of solid phase on reagent strip at this compound on the reagent strip; Detector carries out quantitatively thing to be checked by the power that detects fluorescence signal, and fluorescence signal intensity has reflected captive CRP concentration.

Quantitative fluorescence analysis instrument deft design provided by the invention, be easy to carry, friendly interface, but fast quantification detects omnidistance c reactive protein, glycosylated hemoglobin, microdose urine protein, prostate specific antigen, alpha-fetoprotein, carcinomebryonic antigen, myocardium calcium protein etc.The quantitative Fast Detection Technique of the immunofluorescence that is adopted, detection sensitivity can reach pg/ml.Test item was all finished in 3-15 minute, and the requirement of mass detection can be satisfied in detection speed＜10 in the instrument second/test.

The present invention uses calibration curve to carry out quality control, and the typical curve of test item is stored in the information chip ID card of kit, and the built-in Quality Control of system can be satisfied the requirement of daily Quality Control, guarantees result's accuracy, the CV of whole detection system＜5%.Use the upgrading mode of chip type to carry out the project expanded function.

First detects approximately needs 4 minutes, and each only detects and needs 20 seconds afterwards, can carry out 180 tests in 1 hour, general speed can reach at least 50-60 test/hour, skilled speed can accomplish 80 tests/hour more than.The key of decision detection speed is the time of instrument autoscan.

Because employed starting material of reagent card of each batch and technology is different, curve is also different, therefore when each batch reagent card uses, need a calibration curve, calibration curve is stored in the calibration curve chip, chip is inserted the reagent card slot of machine, with reagent card, change chip, read chip and obtain typical curve.

Immunofluorescence detector among the present invention preferably is applicable to the detection of the omnidistance C-reactive protein of immunofluorescence detection by quantitative, glycosylated hemoglobin, microalbumin, prostate specific antigen, carcinomebryonic antigen, cardiac troponin.The present invention is applicable to vitro detection, go for medical institutions central laboratory, door the quick and quantitative determination system of Laboratory, clinical department and other medical services point, MEC.

With fluorescent latex as tracer, be applied to antigen-antibody reaction, be used for the detection of CRP, FOP etc. by the fluorescent quantitation detecting instrument, the fluorescent quantitation testing result is to judging the infective power of the state of an illness and instructing antiviral treatment that crucial meaning is arranged, state of an illness information more accurately can be provided, and testing result can preserve with data mode, sets up patient's state of an illness database, convenient inquiry in the future.The principal feature that fluorescent quantitation detects is simple to operate quick, does not need professional, highly sensitive, accurately reliable.Be specially adapted to vast grass-roots unit, hospital, field work personnel and the detection of time in enormous quantities and big immunity generaI investigation and do not have professional's environment on the scene, but test item is many, the scope of application is extensive.

The repeated coefficient of variation of the present invention: detect with a standard items,parallel detection 10 times calculates the CV value and answers≤15%.

The stability coefficient of variation: detect with a standard items, reagent strip was put into the detector follow-on test 30 minutes, and test interval is 1 minute, calculated the CV value and answered≤8%.

The above is preferred embodiment of the present invention only, and protection scope of the present invention is not limited thereto, and anyly all belongs within the protection domain of the present invention based on the equivalent transformation on the technical solution of the present invention.

Claims (10)

1. a fluorescent quantitation detector is characterized in that, it comprises excitation source module, photoelectric conversion module, control analysis module and software systems, and described excitation source module comprises the first laser module assembly and the second laser module assembly; The first laser module assembly is used for the detection zone launching excitation light bundle A to reagent strip, through reflecting fluorescence signal C; The second laser module assembly is used for the code area launching excitation light bundle B on reagent strip, through reflecting laser signal D; Described photoelectric conversion module is used to receive the fluorescence signal C and the laser signal D of reflection and carries out the photosignal conversion; Described control analysis module comprises main circuit, motor, display screen, and the control analysis module is used to handle the electric signal that receives photoelectric conversion module output and handles and outputs to display screen and show; Described fluorescent quantitation detector is provided with reagent strip slot and the ID card slot that is used to place reagent strip and calibration chip id card; This motor is used to drive reagent strip and moves.

2. fluorescent quantitation detector as claimed in claim 1 is characterized in that, this fluorescent quantitation detector also comprises a microprinting machine, is used to print testing result.

3. fluorescent quantitation detector as claimed in claim 1 is characterized in that, this photoelectric conversion module comprises a signal compensation circuit, reduces the generation of undesired signal.

4. fluorescent quantitation detector as claimed in claim 1 is characterized in that, the wavelength of the excitation beam A that the described first laser module assembly sends is for being 470nm, and when reagent strip was shone, the reflected fluorescent light C wavelength that fluorescent material sends was 530nm.

5. fluorescent quantitation detector as claimed in claim 1 is characterized in that, described display screen adopts the touching display screen device, is used for instruction input and data presentation.

6. fluorescent quantitation detector as claimed in claim 1 is characterized in that, there is the code area on described matching used reagent strip surface, and this code area is read by the second laser module assembly and import main circuit after the photosignal conversion, for distinguishing different test items.

7. fluorescent quantitation detector as claimed in claim 1 is characterized in that, stores the calibration curve of respective batch reagent strip in the described calibration chip id card.

8. fluorescent quantitation detector as claimed in claim 7 is characterized in that, in the described control analysis module quality control module is arranged, and is used for reading the calibration curve of calibration chip id card, and carries out data according to calibration curve and judge.

9. fluorescent quantitation detector as claimed in claim 1 is characterized in that, also comprises a built-in storage unit, can store up-to-date a plurality of testing results.

10. fluorescent quantitation detector as claimed in claim 1 is characterized in that, described software systems comprise initialization module, start calibration module, system parameter setting module, bar code acquisition module and test sample analysis module.

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