CN201277975Y - Micro-fluidic chip special for syphilis diagnosis - Google Patents
Micro-fluidic chip special for syphilis diagnosis Download PDFInfo
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- CN201277975Y CN201277975Y CNU2008201757042U CN200820175704U CN201277975Y CN 201277975 Y CN201277975 Y CN 201277975Y CN U2008201757042 U CNU2008201757042 U CN U2008201757042U CN 200820175704 U CN200820175704 U CN 200820175704U CN 201277975 Y CN201277975 Y CN 201277975Y
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Abstract
The utility model relates to a micro-fluidic chip specially used for diagnosing syphilis, which belongs to the analyzing and testing field. Related syphilis diagnosis technology requires rapid and cheap syphilis diagnosis, so the utility model provides a micro-fluidic chip for simultaneously detecting and diagnosing syphilis rapidly with various specific syphilis antibodies. The micro-fluidic chip is characterized by being internally provided with capillary channels which are served as four micro-channels connected in parallel, and four working electrodes are respectively arranged in the four micro-channels; each working electrode comprises a metal electrode and a golden size sensitive film adhered to the surface of the metal electrode and enwrapped in a specific syphilis antigen, and the golden size sensitive films adhered to the surfaces of the working electrodes are enwrapped in four different specific syphilis antigens TP0684, TP0453, TP0821 and TP0319. The micro-fluidic chip is favorable for improving the syphilis diagnosis efficiency.
Description
Technical field
The utility model relates to a kind of controller used in syphilis diagnosis micro-fluidic chip special, belongs to the analytical test field.
Background technology
Syphilis (Syphilis) be by microspironema pallidum (T.pallidum, a kind of sexually transmitted disease that TP) causes (Sexually transmitted disease, STD).Microspironema pallidum only infects the mankind under natural situation, and the people is the unique infection sources of syphilis, main trafficability characteristic contact transmission.Also can pass through the placenta vertical transmission in addition, cause fetal congenital syphilis.A few peoples also can infect through blood transfusion.Since the eighties, syphilis is revivable once more in China, according to national venereal disease control center statistics, China's syphilis incidence of disease annual growth reaches 52.7%, and the congenital syphilis and the neurolues incidence of disease increase obviously, the trend that rises is gradually also arranged in the whole world, syphilis not only become international serious society and public health problem, and be one of principal disease of harm population health.Therefore, select simple, fast, hypersensitivity and specificity diagnostic method and reagent, to the syphilis early diagnosis, find the infection sources, control syphilis is propagated significant.
Controller used in syphilis diagnosis method commonly used at present has etiological examination, non-syphilitic leptospira antigen serum test, the test of microspironema pallidum hemagglutination (TPHA), polymerase chain reaction (PCR) and TP quick detection test paper bar etc.The principle of etiological examination is to get the exudate of patient's infringement or the tissue fluid that lymphnode puncture obtains, observe spirochetal characteristic morphologic and mode of motion, in theory early syphilis there is higher recall rate, but the restriction (comprising prescription on individual diagnosis medication, high-quality fluorescent microscope, reagent and technology etc.) that is subjected to condition is also more, and therefore actual recall rate is not high.The principle of non-syphilitic leptospira antigen serum test is the serology primary dcreening operation test commonly used of current diagnosis syphilis, though in a lot of documents, all mention non-syphilitic leptospira antigen serum test simple and easy, quick, be easy to observations, the screening serum that is suitable for a large amount of crowds, but susceptibility, specificity are relatively poor, biologic fale-positive and false negative reaction appear sometimes, cause omission and flase drop easily, so be not suitable for the diagnosis of primary syphilis, congenital syphilis, neurolues.Though its susceptibility of TPHA and specificity height mainly are used to the syphilis confirmed diagnosis test, and observation of curative effect is had little significance.Though TP quick detection test paper bar has overcome defectives such as length detection time, reagent costliness, domestic temporarily do not have its sensitivity of play-by-play and specificity effect, so be not useable for confirmed diagnosis test.The PCR method is newer diagnostic method, and it can carry out early stage antidiastole to genital ulcer, distinguishes syphilis, genital herpes and chancroid.But the PCR method of current diagnosis syphilis still exist primer non-specific, to shortcomings such as susceptibility such as serum/whole blood are low.And when the skin damage begins healing, secretion minimizing, serological test positive reaction, adopt PCR method just improper.In addition, PCR method is subject to materials such as tissue in the sample and cell fragment and suppresses, and causes the PCR false negative result, so detect should not be as declaring more for PCR.
Detection methods such as non-syphilitic leptospira antigen serum test, TPHA and PCR and TP quick detection test paper bar all belong to immunoassay.Above immunoassay itself has very strong selectivity, to the early diagnosis of infectious disease, judges that all there is important meaning aspects such as curative effect and prognosis.Yet the detection of serious infectious diseases is very complicated, and traditional immunoassay only detects at monospecific antibody usually.Because a kind of diagnosis antibody can appear at different pathogenic bacteria, or different diagnosis antibody can occur simultaneously in same pathogenic bacteria, therefore, carry out multiple efficiently immune analysis method (SMIAs), the kind and the quantity of, quantitative response various pathogenic bacteria antibody qualitative truly from the angle of biological correlativity, in early days prevention and the multiple infectious disease of joint inspection, judge that aspects such as disease carrying germ development degree, O﹠A result of treatment all have important medical value.
Electrochemical immunosensor has unique advantages such as automaticity height, cheapness, easily preparation, rapid sensitive, cooperates immunoreactive efficiently single-mindedly, is fit to very much the analyser of the multiple immunoassay detection of structure serious infectious diseases.The micro-fluidic chip (lab-on-chip) that the title of " life science integrated circuit " is arranged is the desirable technique that makes up microminiaturized, integrated electrochemical immunosensor.The micro-fluidic chip technology be with sampling, pre-service, add reagent, reaction, separation, detection etc. be integrated in finish on the microchip one in front of the door along technology, have analysis speed fast, contain much information, reagent consumption less, advantage such as pollution is little, operation cost is low, instrument is easy to use.And be fit to very much suitability for industrialized production.On behalf of the 21 century analytical instrument, microflow control technique move towards microminiaturized, integrated developing direction.The micro-fluidic Amperometric Detection Coupled chip that the serious infectious diseases that are suitable for structure detect has following advantage: (1) is easy to integrated a plurality of sense channels and detecting electrode on chip by micro-processing technology, accomplishes that multiple target objects analyzes the technical feasibility height simultaneously.(2) by the micro-flowing injection technology, sample flows in pipeline and has guaranteed that electrode surface upgrades constantly, and it is easily contaminated better to have overcome electrode, causes false positive rate height, problem such as collimation difference as a result.(3) microfluidic sensing device upper pipeline diameter has only μ m level, and the reaction tank volume also has only μ L level, and required reagent dosage is few during analysis, and analyte arrives the diffusion length of electrode surface lacks, therefore can significantly reduce the incubation time, realizes the fast joint inspection target.(4) the microfluidic sensing device is many based on the optical detection system at present, and the expensive volume of instrument is big, fully integrated being difficult to, and also it is very high to detect cost, has limited its popularization.And adopting Electrochemical Detection in the sensor, volume is little, automaticity is high, cost is low.
At present, in the micro-fluidic chip technical field, utilize that the multiple specific antigen of syphilis detects simultaneously, the correlation technique and the method for the multiple syphilis antibody of quick diagnosis are not appeared in the newspapers as yet.
The utility model content
Technical problem to be solved in the utility model is, in the so total technological frame of micro-fluidic chip technology, develop a kind of can utilize the multiple specific antigen of syphilis come to multiple syphilis antibody detect simultaneously, the micro-fluidic chip special and the detection method thereof of quick diagnosis.
The utility model solve the technical problem by following scheme, the device that this scheme provides is a kind of controller used in syphilis diagnosis micro-fluidic chip special, the structure of this micro-fluidic chip comprises that applying is installed in two plate objects together, described two plate objects are respectively the cover plate of micro-fluidic chip and the substrate of micro-fluidic chip, on the position of the mutual applying between described two plate objects, be equiped with capillary channel, and, four little liquid storage tanks, one end of capillary channel via the manifold shape microchannel respectively with the little liquid storage tank of wherein three UNICOM, the other end of capillary channel and a remaining little liquid storage tank UNICOM, and, be installed in the described capillary channel working electrode on the diverse location in regular turn respectively and to electrode and contrast electrode, described working electrode by metal electrode and be attached to embedding on the described metal electrode gold size sensitive membrane of syphilis specific antigen constitute, this case improvements are, the structure of described capillary channel is structure in parallel, the described capillary channel that is structure in parallel is made of four minim channel parallel connections, and, the quantity of described working electrode is four, the installation position of these four working electrodes lays respectively in described four minim channels, and, specific antigen material in its top layer gold size sensitive membrane structure of these four working electrodes is respectively that these four kinds of antigenic substances are respectively syphilis specific antigen TP0684 to four kinds of syphilis antigenic substances of syphilis antibody energy specificity combination, TP0453, TP0821 and TP0319.Described gold size sensitive membrane is that shitosan gold size solution and syphilis specific antigenic solution are fully mixed, and its drying and forming-film is formed.Described gold size sensitive membrane has been included as fixing above-mentioned each syphilis specific antigen and has introduced wherein complementary medium, and described complementary medium is wherein a kind of of shitosan, cellulose acetate, gelatin or their potpourri for example.Relate to some kinds of electrodes in the structure of this case micro-fluidic chip, described electrode is minute sized electrode, the shape of each electrode wherein all can be selected arbitrarily shape, and described selected arbitrarily shape is square shape sheet shape, rectangle sheet, strip or round sheet or the like for example.Relate to several little liquid storage tanks in this case microfluidic chip structure, described little liquid storage tank is little pond or the micro-capsule that is used for transitional liquid storage, its shape of miniature inner chamber of the little liquid storage tank of wherein each all can be arbitrarily selected shape, for example miniature cylindrical cavity shape of described cavity shape, miniature square column type cavity-like, miniature oblong cavity shape or miniature spherical hollow space shape or the like.This case device can further include some annexes certainly, and described annex is multiple tracks electrochemical workstation and microfluidic pump or the like for example, and the art-recognized meanings of the art-recognized meanings of described multiple tracks electrochemical workstation and microfluidic pump is known.Each working electrode that relates in this case microfluidic chip structure and to electrode and contrast electrode etc., can get lines crossed via corresponding special use is respectively connecting with the corresponding interface of described multiple tracks electrochemical workstation.It is to be used for the private cable that each the corresponding interface with each described electrode and described multiple tracks electrochemical workstation is coupled to each other that described special use is being got lines crossed.Described microfluidic pump is exclusively used in and drives micro liquid and flow, described microfluidic pump can with the described little liquid storage tank of selected any one as required UNICOM.
The specific implementation path of this case micro-fluidic chip can have multiple, selects the one detailed annotation as follows:
One, superpolymer (PC: the preparation of the little gold electrode of electroless plating on cover plate polycarbonate)
1, is photomask with quartz/chromium plate, as light source the selective area on PC cover plate surface carried out radiation with the ultraviolet light of selecting wavelength.
2, adopt hexane diamine as aminating agent, 1-[3 ,-(N-N-dimethyl amine) propyl group]-3-hexyl carbodiimide hydrochloride is that coupling agent carries out aminating reaction to the PC cover plate through the selective area irradiation.
3, will react in the PC cover plate immersion gold chloride through the selective area aminating reaction.
4,, will reduce in its immersion sodium borohydride solution again with after the distilled water cleaning.
5, with after the distilled water cleaning, again with ultrasonic cleaning 30Min in the potassium rhodanate solution of its immersion 0.5mol/L.
6, will contain in the plating bath of gold ion, complexing agent and reductive agent formaldehyde through the PC cover plate immersion of cleaning, react 1h down in 45-60 ℃.
7, the PC cover plate that will be coated with little gold electrode is put into the about 3h of baking oven annealing in process, promptly obtains the many little gold electrode cover plates of superpolymer of the present invention.The long 3mm of wherein little gold electrode, wide 3mm, thickness is about 0.1mm.Spacing is about 3mm each other.
Two, the superpolymer (PC: the making of substrate polycarbonate) that has the microchannel
1, makes silicon mold
Draw floor layout, the pattern line width is about 25-100 μ m, adopts the output of high-resolution laser photocomposing system on transparent film, promptly obtains mask.
On substrate surface, deposit one deck silicon nitride film as sacrifice layer by chemical vapour deposition technique, on this substrate, cover one deck light-sensitive emulsion, in baking oven, handle 15-20min for 60 ℃ by the rotation film coating technique; Mask is covered on the substrate, the principle by the exposure imaging with the image transfer on the mask to the optical cement layer of substrate surface; Method by dry etching is processed into the spatial structure with certain depth with the planar figure on the optical cement layer.Can obtain the silicon mold of lobed passage.The silicon mold for preparing is used H successively
2O
2: H
2SO
4Solution (volume ratio is 1: 4), acetone and distilled water clean, to remove its surperficial oxide.
2, adopt pressure sintering to duplicate the microchannel
The PC sheet cut into be about 6cm, the size of wide 4cm.Ultrasonic cleaning, natural airing is heated to the PC sheet about softening temperature (110 ℃) in hot-press arrangement, applies certain pressure then on silicon mold, and continues about 1min, can suppress the microchannel with the silicon mold complementation on the PC sheet.Then, formpiston and the PC substrate that is carved with the microchannel are cooled off the back demoulding together, just obtain required microchannel.
3, on cover plate, make little liquid storage tank
On cover plate, adopt diamond to get out the sample liquid pool, buffer pool, sample waste liquid pool, four little liquid storage tanks such as buffering waste liquid pool.
Three, the PC substrate that will be coated with the PC cover plate of little gold electrode and have a microchannel is handled in the cryogenic oxygen plasma atmosphere totally
Four, with embedding the gold size sensitive membrane of four kinds of syphilis specific antigens fit in corresponding little metal working respectively and do on the electrode.
Five, chip involution
Aim at little gold electrode cover plate of superpolymer and the superpolymer substrate that has the microchannel at microscopically, with this substrate with cover plate is fixed and clamp with two cover glasses, be placed in the baking oven and under uniform temperature, be incubated 15min, can obtain required four-way PC organic glass sheet micro-fluidic chip.
Six, the PC sheet that step 4 is finished is respectively at working electrode, reference electrode and utilize elargol to connect copper sheet as lead to electrode end, treats that elargol seals up hot melt adhesive after dried fully again with fixing copper sheet, promptly finishes the making of whole four-way micro-fluidic chip.
Its relevant each operating parameter of the above-mentioned specific implementation path of this case micro-fluidic chip of detailed annotation can be adjusted according to actual needs.
The using method of this case micro-fluidic chip:
Employing adds Micropump and drives the steady flow in the capillary channel of four-way micro-fluidic chip of liquid stream, utilizes the four-way electrochemical analytical instrument respectively four kinds of controller used in syphilis diagnosis antibody to be detected.
The concrete detection of this case micro-fluidic chip uses step as follows:
1, add blood serum sample liquid in the microtubule road, adding under the Micropump driving, various syphilis antibody molecules are by the corresponding syphilis specific antigen capture of gold size sensitive membrane embedding on the electrode surface in each passage.
2, the ELIAS secondary antibody that adds horseradish peroxidase-labeled forms immune complex by sandwich method desmoenzyme mark " two is anti-".
3, adopt the hyperchannel electroanalyzer, add electron mediators such as catechol, adopt ampere method detection " two is anti-" to go up enzyme and cause H
2O
2The electric current of reduction changes, and obtains the kind and the content of each analyte thus.
4, the result is carried out analysis-by-synthesis, syphilis antibody is carried out comprehensive diagnos.
The utility model has the advantages that, integrated four working electrodes that are coated with four kinds of syphilis specific antigenic substances respectively on a micro-fluidic chip, these four working electrodes are respectively at four kinds of feature diagnosis antibody that detect syphilis, this case micro-fluidic chip is a kind of controller used in syphilis diagnosis micro-fluidic chip special that can utilize the multiple specific antigen of syphilis to detect simultaneously, and the design feature of its integrated structure has determined the use of this chip to help to accelerate controller used in syphilis diagnosis speed, improves accuracy rate of diagnosis, reduced the controller used in syphilis diagnosis expense.
Description of drawings
Fig. 1 is this case micro-fluidic chip embodiment organigram, comprises described annex among the figure.
Among the figure, the 1st, the microfluidic pump, 2,4,5,14 is respectively four little liquid storage tanks that installation position is different, the 3rd, the manifold shape microchannel, the 6th, the cover plate of micro-fluidic chip, the 7th, the substrate of micro-fluidic chip, the 8th, the special use that connects working electrode and multiple tracks electrochemical workstation is being got lines crossed, the special use that is used to connect working electrode and multiple tracks electrochemical workstation that shows four homotypes in this illustration is altogether being got lines crossed, the 9th, the multiple tracks electrochemical workstation, the 10th, be installed in the working electrode in the minim channel with syphilis specific antigen TP0684, the 11st, be installed in the working electrode in the minim channel with syphilis specific antigen TP0453, the 12nd, be installed in the working electrode in the minim channel with syphilis specific antigen TP0821, the 13rd, be installed in the working electrode in the minim channel with syphilis specific antigen TP0319, the 15th, contrast electrode, the 16th, to electrode, the 17th, connect the special use of electrode and multiple tracks electrochemical workstation is being got lines crossed, the 18th, the special use that connects contrast electrode and multiple tracks electrochemical workstation is being got lines crossed.
Embodiment
In this case embodiment that Fig. 1 showed, the structure of this micro-fluidic chip comprises that applying is installed in two plate objects together, the cover plate 6 that two plate objects are respectively micro-fluidic chips and the substrate 7 of micro-fluidic chip, on the position of the mutual applying between two plate objects, be equiped with capillary channel, one end of capillary channel via manifold shape microchannel 3 respectively with sample liquid pool 2 and buffer pool 4 and sample waste liquid pool 5 UNICOMs, the other end of capillary channel and remaining buffering waste liquid pool 14 UNICOMs, and, be installed in the described capillary channel working electrode on the diverse location in regular turn respectively and to electrode 16 and contrast electrode 15, working electrode by metal electrode and be attached to embedding on the described metal electrode gold size sensitive membrane of syphilis specific antigen constitute, the structure of capillary channel is structure in parallel, this capillary channel that is structure in parallel is made of four minim channel parallel connections, and, the quantity of described working electrode is four, the installation position of these four working electrodes lays respectively in described four minim channels, and these four kinds of antigenic substances are respectively syphilis specific antigen TP0684, TP0453, TP0821 and TP0319.These four working electrodes are respectively to be installed in the working electrode 10 with syphilis specific antigen TP0684 in the first tributary in parallel minim channel and to be installed in the working electrode 11 with syphilis specific antigen TP0453 in the second tributary in parallel minim channel and to be installed in the working electrode 12 with syphilis specific antigen TP0821 in the 3rd tributary in parallel minim channel and to be installed in the working electrode 13 with syphilis specific antigen TP0319 in the 4th tributary in parallel minim channel.Comprise multiple tracks electrochemical workstation 9 and microfluidic pump 1 in this illustration as described annex, contrast electrode 15 can got lines crossed via special use and 18 connected with the corresponding interface of multiple tracks electrochemical workstation 9, can get lines crossed via special use to electrode 16 and 17 to connect with the corresponding interface of multiple tracks electrochemical workstation 9, working electrode 10,11,12 and 13 can be is respectively getting lines crossed via four homotype special uses and 8 to connect with the corresponding interface of multiple tracks electrochemical workstation 9, as the microfluidic pump 1 of annex can with the little liquid storage tank of selected any one UNICOM as required, in this example, microfluidic pump 1 and little liquid storage tank 4 UNICOMs.
Claims (1)
1. controller used in syphilis diagnosis micro-fluidic chip special, the structure of this micro-fluidic chip comprises that applying is installed in two plate objects together, described two plate objects are respectively the cover plate of micro-fluidic chip and the substrate of micro-fluidic chip, on the position of the mutual applying between described two plate objects, be equiped with capillary channel, and, four little liquid storage tanks, one end of capillary channel via the manifold shape microchannel respectively with the little liquid storage tank of wherein three UNICOM, the other end of capillary channel and a remaining little liquid storage tank UNICOM, and, be installed in the described capillary channel working electrode on the diverse location in regular turn respectively and to electrode and contrast electrode, described working electrode by metal electrode and be attached to embedding on the described metal electrode gold size sensitive membrane of syphilis specific antigen constitute, it is characterized in that, the structure of described capillary channel is structure in parallel, the described capillary channel that is structure in parallel is made of four minim channel parallel connections, and, the quantity of described working electrode is four, the installation position of these four working electrodes lays respectively in described four minim channels, and, specific antigen material in its top layer gold size sensitive membrane structure of these four working electrodes is respectively that these four kinds of antigenic substances are respectively syphilis specific antigen TP0684 to four kinds of syphilis antigenic substances of syphilis antibody energy specificity combination, TP0453, TP0821 and TP0319.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU2008201757042U CN201277975Y (en) | 2008-10-21 | 2008-10-21 | Micro-fluidic chip special for syphilis diagnosis |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU2008201757042U CN201277975Y (en) | 2008-10-21 | 2008-10-21 | Micro-fluidic chip special for syphilis diagnosis |
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| CN201277975Y true CN201277975Y (en) | 2009-07-22 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNU2008201757042U Expired - Fee Related CN201277975Y (en) | 2008-10-21 | 2008-10-21 | Micro-fluidic chip special for syphilis diagnosis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106248939A (en) * | 2015-06-11 | 2016-12-21 | 宁波大学 | A kind of multichannel syphilis diagnosis micro flow control chip device |
-
2008
- 2008-10-21 CN CNU2008201757042U patent/CN201277975Y/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106248939A (en) * | 2015-06-11 | 2016-12-21 | 宁波大学 | A kind of multichannel syphilis diagnosis micro flow control chip device |
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| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090722 Termination date: 20111021 |