CN103760338B - Quantitative detection method using reverse immunochromatography reaction, kit and external equipment - Google Patents
Quantitative detection method using reverse immunochromatography reaction, kit and external equipment Download PDFInfo
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- CN103760338B CN103760338B CN201410036880.8A CN201410036880A CN103760338B CN 103760338 B CN103760338 B CN 103760338B CN 201410036880 A CN201410036880 A CN 201410036880A CN 103760338 B CN103760338 B CN 103760338B
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- 238000006243 chemical reaction Methods 0.000 title claims abstract description 42
- 238000001514 detection method Methods 0.000 title claims abstract description 41
- 238000003317 immunochromatography Methods 0.000 title claims abstract description 17
- 238000005406 washing Methods 0.000 claims abstract description 40
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 35
- 239000007788 liquid Substances 0.000 claims abstract description 26
- 230000035484 reaction time Effects 0.000 claims abstract description 4
- 239000006210 lotion Substances 0.000 claims description 37
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 claims description 27
- 229940005654 nitrite ion Drugs 0.000 claims description 27
- 238000002347 injection Methods 0.000 claims description 18
- 239000007924 injection Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 18
- 238000012360 testing method Methods 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 12
- 238000005213 imbibition Methods 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 8
- 239000000376 reactant Substances 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 230000002093 peripheral effect Effects 0.000 claims description 3
- 239000000126 substance Substances 0.000 abstract description 3
- 239000012528 membrane Substances 0.000 abstract 2
- 230000003287 optical effect Effects 0.000 abstract 2
- 239000008267 milk Substances 0.000 description 31
- 210000004080 milk Anatomy 0.000 description 31
- 235000013336 milk Nutrition 0.000 description 31
- 150000001875 compounds Chemical class 0.000 description 14
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 9
- 239000004816 latex Substances 0.000 description 9
- 229920000126 latex Polymers 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 210000002700 urine Anatomy 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 206010029719 Nonspecific reaction Diseases 0.000 description 1
- 208000002151 Pleural effusion Diseases 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- -1 nitrite ions Chemical class 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 235000019371 penicillin G benzathine Nutrition 0.000 description 1
- 229940056360 penicillin g Drugs 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
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- Analytical Chemistry (AREA)
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a detection method, a kit and external equipment applying reverse immunochromatography reaction, which accurately control the reaction amount and the reaction time of a sample to be detected by using the external equipment and the kit, thereby realizing the reverse immunochromatography reaction; a chromatographic membrane is arranged in the kit, and the middle part of the chromatographic membrane is provided with a control area and a detection area; when the external equipment detects that the sample chromatography reaches the designated position, the sample chromatography is completed, the external equipment is automatically controlled to inject washing liquor and color development liquid or luminous liquid into the kit in sequence, the external equipment controls the imbibing pump to act on the kit, so that the injected liquid carries out reverse chromatography, reacts with substances in the contrast area and the detection area in sequence, and finally generates color change or visible light, and finally the external equipment identifies optical signals of the contrast area and the detection area and compares the optical signals of the contrast area and the detection area with a built-in standard curve to obtain a quantitative detection result.
Description
Technical field
The present invention relates to a kind of quantitative detecting method and kit and the external unit of applying the reaction of reverse immunochromatography, particularly a kind of apply the reaction of reverse immunochromatography quantitative detecting method and kit and external unit and kit used thereof and external unit.
Background technology
At present, the detection method for biological specimen is a lot, and wherein IDEXX company employs a kind of special kit, and washing lotion and nitrite ion are stored in kit by it, and the top of washing lotion and nitrite ion arranges chromatographic film.Its specific works principle (being detected as example with milk) is: after milk sample being put into Reagent Tube heating 5 minutes (45 DEG C), in instillation kit, chromatographic film utilizes capillarity milk sample chromatography to be moved to check point in chromatographic film and control point, material in milk sample and check point and control point is reacted, then after observing milk sample chromatography to certain position (detection window place), the side of kit is pressed, this side due to kit is provided with the kapillary that two connect washing lotion pond and chromatographic film and develop the color liquid pool and chromatographic film respectively, so after kit side presses, the washing lotion pond of kapillary puncture through seal and colour developing liquid pool, and washing lotion and nitrite ion are drawn to chromatographic film place, then carry out chromatography and move to check point and control point, wash-out and colour developing are carried out to check point and control point, the color of finally observing check point and control point can draw testing result.The structure of its kit as shown in Figure 1.
But in this, kit is owing to storing washing lotion and nitrite ion, and inner structure is comparatively complicated, and therefore volume is comparatively huge, carries inconvenience.In addition, because general washing lotion and nitrite ion need to keep lower temperature, kit needs stored refrigerated all the time in long-distance transport process, and its transportation cost is higher.And, because the needs of the washing lotion in each kit and nitrite ion realize being encapsulated in kit, its consumption must be unified just can be convenient to large-scale production, in order to the detection of most sample can be adapted to, the encapsulation amount of washing lotion and nitrite ion generally need be greater than actual use amount, so can cause very large waste.In addition, because the program is substantially all utilize the capillary action of chromatographic film self to make liquid circulate, so the requirement of kit to chromatographic film is higher, need its inside to have abundant kapillary, and need to keep away from moisture in preserving.Therefore the chromatographic film that IDEXX company is used is also generally special, its production cost and retain costs higher.It will be further appreciated that, the program can only qualitative detection sample, can not export quantitative result, so can only judge negative positive, can not export the concentration value determined.
Summary of the invention
The object of the invention is to, a kind of quantitative detecting method and kit and the external unit of applying the reaction of reverse immunochromatography are provided.It can solve that the kit existed in prior art cannot export quantitative result, carries inconvenience, transportation cost and retain costs is high and the problem such as washing lotion and nitrite ion waste.
Technical scheme of the present invention: the quantitative detecting method applying the reaction of reverse immunochromatography, is characterized in: use external unit and kit accurately to control the reacting dose of sample to be tested and reaction time, thus realize the reaction of reverse immunochromatography, be provided with chromatographic film in this agent box, in the middle part of chromatographic film, be loaded with control zone and surveyed area, when external unit detects that sample chromatography arrives assigned address, sample chromatography completes, washing lotion and nitrite ion or luminescent solution are successively injected kit by automatic control external unit, peripheral equipment control suction pumps acts on kit, the liquid of injection is made to carry out reverse chromatography, and successively carry out reacting (immune response and/or chemical reaction) with the material in control zone and surveyed area, and the last color that produces changes or produces visible ray, last external unit is by identifying the light signal of control zone and surveyed area, and quantitative testing result can be drawn after the light signal of control zone and surveyed area and built-in typical curve being contrasted.
Built-in typical curve is stored in advance in external unit, and it is the weak homologous thread (can sum up matching according to many experiments to form) with test substance concentration in sample of a bars light intensity.Therefore namely may correspond to after recording light signal strength and obtain concentration information.
In the quantitative detecting method of the reverse immunochromatography reaction of above-mentioned application, the concrete steps of pattern detection comprise:
1. in the well of the wherein side of kit, carry out quantitative application of sample, sample chromatography under the capillary action of chromatographic film moves to control zone and surveyed area, and reacts with the material in control zone and surveyed area;
2. when external unit detects that sample to be tested chromatography is to indication position, external unit first injects washing lotion again in the liquid injection hole of the opposite side of kit, and on well, use suction pumps to aspirate, and makes the reverse chromatography of washing lotion move to well;
3. then in liquid injection hole, add nitrite ion or luminescent solution, suction pumps works on simultaneously, makes nitrite ion or luminescent solution and washing lotion carry out chromatography in the same way, then chromogenic reaction occurs for control zone and surveyed area;
4. the last light signal (comprise color signal) of external unit to surveyed area and control zone identifies automatically, after final external unit compares to the power of signal and built-in typical curve, draw quantitative testing result, and testing result is shown to user.
In the quantitative detecting method of the reverse immunochromatography reaction of aforesaid application, described surveyed area and control zone are for being loaded with point or the line of reactant (latex/magnetic bead/antibody).
For the kit of preceding method, be characterized in: the chromatographic film comprising bar shaped, chromatographic film is coated with box body, and the middle part of box body is provided with detection window, and the chromatographic film of detection window position is provided with control zone and surveyed area; The box body of described detection window both sides offers respectively the well for adding sample and the liquid injection hole for adding washing lotion and nitrite ion or luminescent solution, liquid injection hole is also for detecting the chromatography position of sample; The reactant reacted with sample is loaded with respectively in described control zone and surveyed area.
For the external unit of preceding method, be characterized in: at least comprise a floor component, floor component is provided with bracket component, is provided with for holding kit and slidably passing in and out the board supporting plate of external unit, bracket component is fixed with imbibition parts and refill unit below bracket component; Bracket component is also fixed with barcode detection parts, image-detection component; Also comprise display unit and control the control mainboard of each parts.
In aforesaid external unit, also comprise IC-card read-write parts, heater block and fluorescent scanning detection part.
Compared with prior art, the present invention by an external unit and more easy kit with the use of to realize testing goal.By the monitoring to liquid injection hole, position and the time of sample chromatography can be gone out by automatic analysis by external unit, automatically control the opportunity adding washing lotion and nitrite ion, at the end of surveyed area reaction, add washing lotion, cessation reaction.Compared with prior art item, because machine controls automatically, what prevent overreaction from producing scraps, and improves the precision that reagent detects, the unnecessary error preventing manual operation from causing.Relative to prior art, the interpolation of nitrite ion and washing lotion and the hand over of preservation are to external unit.So kit of the present invention can do very thin, volume is very little, is easy to carry, and fetters without the preservation condition of washing lotion and nitrite ion, and transport, retain costs also reduce a lot.In addition, because external unit can preserve a large amount of nitrite ions and washing lotion, its addition also accurately can be controlled by external unit, and an external unit can repeat to act on different kits, compared with prior art, a large amount of washing lotions and nitrite ion can be saved.And the present invention is also provided with imbibition parts in external unit, in the multiple steps detected, use imbibition parts to act on chromatographic film, liquid is made to carry out chromatography passively, thus the requirement to chromatographic film (being equivalent to chromatographic film of the prior art) performance own is reduced, enhance washability simultaneously, reduce nonspecific reaction.The present invention is when carrying out pattern detection, kit is only needed to insert external unit, again sample to be tested is added external unit, other step all can be controlled by external unit automatically, finally can obtain quantitative testing result intuitively in the display system of external unit, the error that the subjective judgement avoiding people produces.
Fig. 1 is the structural drawing of available reagent box;
Fig. 2 is the structural representation of kit of the present invention;
Fig. 3 is the structured flowchart of external unit of the present invention;
Fig. 4 is the side view of Fig. 3.
Being labeled as in accompanying drawing: 1-well, 2-detection window, 3-chromatographic film, 4-liquid injection hole, 5-box body, 6-control zone, 7-surveyed area, 11-floor component, 12-bracket component, 13-imbibition parts, 14-refill unit, 15-image-detection component, 16-board supporting plate, 17-IC card read-write parts, 18-fluorescent scanning detection part, 110-display unit, 111-heater block, 112-controls mainboard.
Below in conjunction with drawings and Examples, the present invention is further illustrated, but not as the foundation limited the present invention.
Embodiment.Apply the detection method of reverse immunochromatography reaction: use external unit and kit accurately to control the reacting dose of sample to be tested and reaction time, thus realize the reaction of reverse immunochromatography, be provided with chromatographic film in this agent box, in the middle part of chromatographic film, be loaded with control zone and surveyed area, when external unit detects that sample chromatography arrives assigned address, sample chromatography completes, washing lotion and nitrite ion or luminescent solution are successively injected kit by automatic control external unit, peripheral equipment control suction pumps acts on kit, the liquid of injection is made to carry out reverse chromatography, and successively carry out reacting (immune response and/or chemical reaction) with the material in control zone and surveyed area, and the last color that produces changes or produces visible ray, last external unit is by identifying the light signal of control zone and surveyed area, quantitative testing result can be drawn after the power of contrast signal and built-in typical curve.
The concrete steps of pattern detection comprise:
1. in the well of the wherein side of kit, carry out quantitative application of sample, sample chromatography under the capillary action of chromatographic film moves to control zone and surveyed area, and reacts with the material in control zone and surveyed area;
2. when external unit detects that sample to be tested chromatography is to indication position, external unit first injects washing lotion again in the liquid injection hole of the opposite side of kit, and on well, use suction pumps to aspirate, and makes the reverse chromatography of washing lotion move to well;
3. then in liquid injection hole, add nitrite ion or luminescent solution, suction pumps works on simultaneously, makes nitrite ion or luminescent solution and washing lotion carry out chromatography in the same way, then chromogenic reaction occurs for control zone and surveyed area;
4. the last light signal (comprise color signal) of external unit to surveyed area and control zone identifies automatically, after final external unit compares to the power of signal and built-in typical curve, and testing result is shown to user.
Described surveyed area and control zone are for being loaded with point or the line of reactant (latex/magnetic bead/antibody).
For the kit of preceding method, as shown in Figure 2: the chromatographic film 3 comprising bar shaped, chromatographic film 3 is coated with box body 5, and the middle part of box body 5 is provided with detection window 2, and the chromatographic film 3 of detection window 2 position is provided with control zone 6 and surveyed area 7; The box body 5 of described detection window 2 both sides offers respectively the well 1 for adding sample and the liquid injection hole 4 for adding washing lotion and nitrite ion or luminescent solution, liquid injection hole 4 is also for detecting the chromatography position of sample; The reactant reacted with sample is loaded with respectively in described control zone 6 and surveyed area 7.
For the external unit of preceding method, as shown in Figure 3 and Figure 4: at least comprise a floor component 11, floor component 11 is provided with bracket component 12, be provided with below bracket component 12 for holding kit and slidably passing in and out the board supporting plate 16 of external unit, bracket component 12 be fixed with imbibition parts 13 and refill unit 14; Bracket component 12 is also fixed with barcode detection parts 19, image-detection component 15; Also comprise display unit 110 and control the control mainboard 112 of each parts.Also comprise IC-card read-write parts 17, heater block 111 and fluorescent scanning detection part 18.Having no special requirements at the position relationship of external unit of each parts, only needing to realize following principle of work can arrange arbitrarily.
The principle of work of external unit is as follows, for milk sample, milk sample is added sample hose, slightly shakes up, then sample hose is put into the heater block 111 of external unit, heating 5min; Sample hose is taken out from heater block 111, milk is poured into the well 1 of kit, sample starts chromatography and moves, kit is put into board supporting plate 16 simultaneously, board supporting plate 16 enters external unit, bar code in barcode detection parts 19 detection kit, then image-detection component 15 detects liquid injection hole automatically, detects sample and whether arrives liquid injection hole; When detecting that sample arrives liquid injection hole, imbibition parts 13 move to well, and start imbibition, and refill unit 14 is started working simultaneously, adds washing lotion; After washing lotion chromatography completes, imbibition parts work on, and add nitrite ion and start chromatography.After nitrite ion chromatography completes, kit moves to detecting position, by image-detection component 15, detect (when adding luminescent solution, fluorescent scanning detection part 18 can be used to detect), coordinate software analysis to provide testing result by controlling mainboard 112, and result is presented on display unit 110.
Now for milk pattern detection, its concrete Cleaning Principle is as follows.
1, explanation.
1) comprise PBP-HRP(PBP-horseradish peroxidase in reaction tube and connect albumen, be called for short PBP-HRP) be connected albumen with R-anti-MIgG-HRP(rabbit against murine immunoglobulin (Ig)-horseradish peroxidase, be called for short RAM-HRP) freeze-dried powder;
2) check point: by solid-phase coating technology, ampicillin Small molecular (Amp) is fixed to micro-nano latex particle and (is called for short: be called for short Latex-Amp Latex), get certain density bag by latex point sample in chromatographic film.
3) control point comprises 2 schemes.
Option A: by solid-phase coating technology mouse IgG (immunoglobulin (Ig)) is fixed on micro-nano latex particle and (writes a Chinese character in simplified form: Latex-MIgG), get certain density bag by latex point sample in chromatographic film.Now, the control antibodies be included in reaction tube is the horseradish peroxidase bond (writing a Chinese character in simplified form R-anti-MIgG-HRP) of rabbit anti-mouse IgG.
Option b: by solid-phase coating technology little mouse-anti PBP protein antibodies is fixed on micro-nano latex particle and (writes a Chinese character in simplified form: Latex-AntiPBP), remove certain density bag by latex point sample in chromatographic film.Now, be included in reaction tube without control antibodies, only have PBP-HRP.
2, course of reaction
1) get 400ul milk sample to be measured, join in reaction tube, mixing, the sample joining external unit adds mouth, 45 DEG C of heating 5min.
The reaction wherein existed:
A) negative milk: any composition of answering in the PBP-HRP and milk melted does not react, and is still present in milk with free PBP-HRP albumen.
B) positive milk: part answers microbiotic reaction (for benzyl penicillin (PenicillinG)) in the PBP-HRP and milk melted, form PenicillinG-PBP-HRP compound, be present in milk, part PBP-HRP still exists in free mode, if high containing PenicillinG concentration in milk to be measured, then free PBP-HRP concentration is low.
C), in option A, R-anti-MIgG-HRP does not participate in reaction.
2), after reaction terminates, join in the well of chromatographic film by 400ul milk, milk to be measured, in the mode of chromatography, flows fast through the position of check point and control point within the regular hour, and arrives the position in washing lotion hole.
In option A, the reaction of wherein check point and control point existence:
A) negative milk: be present in the free PBP-HRP albumen in Fresh Milk, and Latex-Amp reaction in check point, form Latex-Amp-PBP-HRP compound, be fixed on check point.
B) positive milk: PenicillinG-PBP-HRP is owing to reacting, and when flowing through check point, does not react; And part is still with the PBP-HRP that free mode exists, then with Latex-Amp reaction, forms Latex-Amp-PBP-HRP compound and be fixed on check point.Latex-Amp-PBP-HRP compound amount number, depend on the amount of PenicillinG in sample number, in sample, PenicillinG amount is many, then Latex-Amp-PBP-HRP compound amount is few, otherwise otherwise.
C) milk flow control point, R-anti-MIgG-HRP and Latex-MIgG reacts, and forms Latex-MIgG-R-anti-MIgG-HRP, is fixed on control point.
In option b, the reaction of check point and control point:
A) negative milk: be present in the free PBP-HRP albumen in Fresh Milk, respectively with Latex-Amp in check point, the Latex-AntiPBP reaction in control point, forms Latex-Amp-PBP-HRP compound, is fixed on check point; Form Latex-AntiPBP-PBP-HRP compound and be fixed on control point.
B) positive milk: PenicillinG-PBP-HRP is owing to reacting, and when flowing through check point, does not react with latex-Amp; And part is still with the PBP-HRP that free mode exists, then with Latex-Amp reaction, forms Latex-Amp-PBP-HRP compound and be fixed on check point.Latex-Amp-PBP-HRP compound amount number, depend on the amount of PenicillinG in sample number, in sample, PenicillinG amount is many, then Latex-Amp-PBP-HRP compound amount is few, otherwise otherwise.When positive milk flow control point, Latex-AntiPBP and PenicillinG-PBP-HRP and free PBP-HRP reacts, and formation Latex-AntiPBP-PenicillinG-PBP-HRP compound and Latex-AntiPBP-PBP-HRP compound are fixed on control point.In sample, PenicillinG concentration is higher, form PenicillinG-PBP-HRP compound amount more, free PBP-HRP amount is fewer, then in check point, Latex-Amp-PBP-HRP compound amount is few, but control point Latex-AntiPBP-PenicillinG-PBP-HRP is more.
3) wash: when reaction terminates, suction pumps arrives well, and a certain amount of washing lotion joins in washing lotion hole, suction pumps work, washing reaction film
4) develop the color: washing terminates, and adds a certain amount of nitrite ion, suction pumps work in washing lotion hole, colour developing 3min
5) judgement is detected.Image identification system reads check point and control point gray-scale value automatically, and instrument internal arranges typical curve, according to the gray-scale value of check point and control point, and with reference to built-in typical curve, draws quantitative concentration value.
Except the milk in embodiment 1 detects, method and apparatus of the present invention is also applicable to sample that the directed fast immune chromatographic detection method of other utilization carries out detecting and test item, and the sample that in background technology, the kit of IDEXX company can detect and test item all can use method and apparatus of the present invention to detect.The sample that the present invention can detect mainly comprises: blood, serum, blood plasma, urine, Pleural effusions, hydrocrania, cells and supernatant, sputum, throat swab eluent, Fresh Milk, finished product milk, milk powder, animal vegetable tissue homogenate, animal blood, animal blood serum, animals urine and other body fluid of animal etc.The present invention may be used for detecting people and animal blood sample, body fluid sample; pathogen, albumen, other molecule or chemical moleculars etc. are detected in food liquid, animals and plants extract; detecting step is substantially identical with the step in embodiment 1, only needs the material in check point in replacement equipment, control point and corresponding replacement washing lotion and nitrite ion.
Claims (5)
1. the quantitative detecting method of the reverse immunochromatography reaction of application, is characterized in that: use external unit and kit accurately to control the reacting dose of sample to be tested and reaction time, thus realizes the reaction of reverse immunochromatography, be provided with chromatographic film in this agent box, in the middle part of chromatographic film, be loaded with control zone and surveyed area, when external unit detects that sample chromatography arrives assigned address, sample chromatography completes, washing lotion and nitrite ion or luminescent solution are successively injected kit by automatic control external unit, peripheral equipment control suction pumps acts on kit, the liquid of injection is made to carry out reverse chromatography, and successively react with the material in control zone and surveyed area, and the last color that produces changes or produces visible ray, last external unit is by identifying the light signal of control zone and surveyed area, and quantitative testing result can be drawn after the light signal of control zone and surveyed area and built-in typical curve being contrasted.
2. the quantitative detecting method of the reverse immunochromatography reaction of application according to claim 1, it is characterized in that, the concrete steps of pattern detection comprise:
1. in the well of the wherein side of kit, carry out quantitative application of sample, sample chromatography under the capillary action of chromatographic film moves to control zone and surveyed area, and reacts with the material in control zone and surveyed area;
2. when external unit detects that sample to be tested chromatography is to indication position, external unit first injects washing lotion again in the liquid injection hole of the opposite side of kit, and on well, use suction pumps to aspirate, and makes the reverse chromatography of washing lotion move to well;
3. then in liquid injection hole, add nitrite ion or luminescent solution, suction pumps works on simultaneously, makes nitrite ion or luminescent solution and washing lotion carry out chromatography in the same way, then chromogenic reaction occurs for control zone and surveyed area;
4. the last light signal of external unit to surveyed area and control zone identifies automatically, and quantitative testing result is shown to user after comparing to the power of signal and built-in typical curve by final external unit.
3. the quantitative detecting method of the reverse immunochromatography reaction of application according to claim 1 and 2, is characterized in that: described surveyed area and control zone are the point or the line that are loaded with reactant.
4. for the external unit of method described in the arbitrary claim of claims 1 to 3, it is characterized in that: at least comprise a floor component (11), floor component (11) is provided with bracket component (12), bracket component (12) below is provided with for holding kit and slidably passing in and out the board supporting plate (16) of external unit, bracket component (12) is fixed with imbibition parts (13) and refill unit (14); Bracket component (12) is also fixed with barcode detection parts (19), image-detection component (15); Also comprise display unit (110) and control the control mainboard (112) of each parts.
5. external unit according to claim 4, is characterized in that: also comprise IC-card read-write parts (17), heater block (111) and fluorescent scanning detection part (18).
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1786712A (en) * | 2004-12-07 | 2006-06-14 | 盛进生物工程(深圳)有限公司 | Method and system for quantitatively fast level testing flow |
| CN201600367U (en) * | 2009-12-01 | 2010-10-06 | 刘江 | General immunity chromatography card interpretation recorder |
| CN102253195A (en) * | 2011-06-28 | 2011-11-23 | 上海贝西生物科技有限公司 | Full-automatic immunochromatographic analyzer |
| CN102628874A (en) * | 2012-04-28 | 2012-08-08 | 无锡瑞美电子科技有限公司 | Full-automatic colloidal gold detection method and full-automatic colloidal gold detection instrument |
| WO2012141368A2 (en) * | 2011-04-13 | 2012-10-18 | Philosys Co., Ltd. | Test strip analyzing apparatus having wheel ejector |
| CN203672884U (en) * | 2014-01-26 | 2014-06-25 | 杭州海士利生物科技有限公司 | Kit for quantitative detection of sample |
| CN203672893U (en) * | 2014-01-26 | 2014-06-25 | 杭州海士利生物科技有限公司 | External device for quantitative detection of sample |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837823A (en) * | 2005-03-21 | 2006-09-27 | 邓雪梅 | Animal epidemic disease detection kit and detection method thereof |
| EP2677321A1 (en) * | 2012-06-20 | 2013-12-25 | Innogenetics N.V. | Integrated device for nucleic acid hybridizations and immuno-assays |
-
2014
- 2014-01-26 CN CN201410036880.8A patent/CN103760338B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1786712A (en) * | 2004-12-07 | 2006-06-14 | 盛进生物工程(深圳)有限公司 | Method and system for quantitatively fast level testing flow |
| CN201600367U (en) * | 2009-12-01 | 2010-10-06 | 刘江 | General immunity chromatography card interpretation recorder |
| WO2012141368A2 (en) * | 2011-04-13 | 2012-10-18 | Philosys Co., Ltd. | Test strip analyzing apparatus having wheel ejector |
| CN102253195A (en) * | 2011-06-28 | 2011-11-23 | 上海贝西生物科技有限公司 | Full-automatic immunochromatographic analyzer |
| CN102628874A (en) * | 2012-04-28 | 2012-08-08 | 无锡瑞美电子科技有限公司 | Full-automatic colloidal gold detection method and full-automatic colloidal gold detection instrument |
| CN203672884U (en) * | 2014-01-26 | 2014-06-25 | 杭州海士利生物科技有限公司 | Kit for quantitative detection of sample |
| CN203672893U (en) * | 2014-01-26 | 2014-06-25 | 杭州海士利生物科技有限公司 | External device for quantitative detection of sample |
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