CN107192824A - The detection means and its detection method of a kind of information-based food pathogenic infection of rapid qualitative - Google Patents
The detection means and its detection method of a kind of information-based food pathogenic infection of rapid qualitative Download PDFInfo
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The present invention provides a kind of detection means of the information-based food pathogenic infection of rapid qualitative, including food pathogenic infection detector bar, instrument, transmitting device and terminal count device.Wherein, described food pathogenic infection is the one or more in rotavirus, norovirus, adenovirus, helicobacter pylori, large intestine toxic bacillus, salmonella and Li bacillus.Add and " solid-phase coating labelled antibody-determined antigen-labelled antibody " immune complex is formed after sample, its color intensity is directly proportional to labelled antigen concentration to be measured.Such as 10-15 minutes particular detection time, chromatography detector bar (box) is detected by an unaided eye to record and reads result in result or quantitative analysis instrument or is transmitted by transmitting device to the terminal count device.Such as by relevant information by computer or any network equipment, or smart mobile phone, storage sends person under test, such as operating personnel, doctor, or be sent to inspection center to.
Description
Technical field
The invention belongs to biomedical inspection field, more particularly to a kind of information-based food pathogenic infection of rapid qualitative
Detection means and its detection method.
Background technology
Bacterial origin and viral source, enter human body or animal gastrointestinal tract to cause metainfective different clinical conditions by food
Shape：Vomiting, suffers from diarrhoea, heating.Some cause of disease prolonged stay human bodies, such as helicobacter pylori trigger lesion carcinogenic eventually.And these are sick
Original can be infected with large area, cause epidemic disease.Current screening method is enzyme-linked method or luminescence method, PCR, CT, or MRI.Journey
Sequence is complicated, needs professional equipment and professional to operate.Also there are a small amount of flash chromatography method, such as patent in the market
CN101696447A is used to test the test strips of psychrophile in chilled food, can only qualitative detection, it is impossible to make a definite diagnosis and monitor follow-up,
Practical value is relatively low.
The content of the invention
In view of this, it is an object of the invention to provide a kind of detection dress of the information-based food pathogenic infection of rapid qualitative
Put, the device can realize the remote real time monitoring of food pathogenic infection.
Technical scheme is as follows：
The present invention provides a kind of detection means of the information-based food pathogenic infection of rapid qualitative, including
Food pathogenic infection detector bar；
Instrument, is connected by the neck and the food pathogenic infection detector bar that are set on the instrument
Connect, obtain the quantitative testing result of the food pathogenic infection detector bar；
Transmitting device, the quantitative testing result for transmitting the instrument detection；
And terminal count device, the quantitative testing result for reading the transmitting device transmission.
Preferably, in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention, the food
Pathogenic infection detector bar includes sample uptake zone, filtering area, chromatographic zone and suction zones.
Preferably, the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention includes bottom plate, sample
Product uptake zone (bar), filtering area (bar), chromatographic zone (bar) and suction zones (bar) are pasted onto on viscose glue bottom plate, the bottom assembled
Plate；The present invention is preferably first from bottom to top to paste sample uptake zone (the 1st area), filtering area the (the 2nd successively in bottom plate to the assembling
Area), chromatographic zone (the 3rd area), suction zones (the 7th area), paste make on viscose board, during stickup each area overlapping 1~
1.5mm；The present invention does not have special limitation to the material, size and source of the bottom plate, is known using those skilled in the art
Be used for immunoassay strip in bottom plate.In the present invention, the material of the bottom plate is preferably plastics or film condensation material.
To after the bottom plate of assembling, the bottom plate of assembling is cut, immunity test strip is obtained.The present invention does not have to the method for the slitting
It is specifically limited, using slitting method well-known to those skilled in the art.In the present invention, the width of the slitting is preferably
3.0~5.0mm, preferably 4mm.
The immunoassay strip that the present invention is provided includes the sample uptake zone being arranged on the bottom plate.In the present invention, institute
State the material preferably glass fibre of sample uptake zone.The present invention does not have any limitation, ability to the source of the glass fibre
Glass fibre known to field technique personnel.The present invention is pasted to the sample uptake zone in the lower end of bottom plate, will be described
The lower edge of sample uptake zone is concordant with bottom plate lower edge.
In the present invention, the sample that the sample uptake zone absorbs is excrement or the extract of excrement.The source of the excrement
Do not limit, human or animal's excrement.
Preferably, in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention, the chromatography
Area includes immobilized label antibody district and reaction zone, and the reaction zone includes detection zone and check plot.The solid phase labelling antibody
Area is arranged on the bottom plate, above filtering area, and the solid phase labelling antibody district 1~2mm overlapping with filtering area.
In the present invention, the material of the solid phase labelling antibody district is preferably glass fibre.
Preferably, in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention, in addition to set
The reaction zone on the bottom plate is put, the reaction zone is close to the solid phase labelling antibody district, positioned at the solid phase labelling antibody
The top in area, and 1~2mm of the reaction zone and the solid phase labelling antibody area overlapping.In the present invention, the reaction zone
Material is preferably nitrocellulose filter.
Preferably, in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention, the reaction
Area is provided with detection line T and nature controlling line C.The detection line T is located at below nature controlling line C, and the detection line T is close to solid phase labelling
Antibody district, the nature controlling line C is close to suction zones.
The preparation method of food pathogenic infection detector bar of the present invention, comprises the following steps：
Labelled antibody is dissolved in buffer solution, the labelled antibody diluted, the labelled antibody of the dilution is sprayed
On glass fibre membrane, solid phase labelling antibody membrane is obtained after drying, solid phase labelling antibody district is used as；
The reaction zone that coated antibody solution is lined into nitrocellulose film makes detection line, and rabbit anti-mouse igg solution is rule
Nature controlling line is made in the reaction zone of nitrocellulose film, obtains being coated with labelled antibody chromatographic film after drying, is used as reaction zone；
Sample absorbing strip, filtering rod, the solid phase labelling antibody membrane, the coating mark are from bottom to top pasted successively in bottom plate
Remember after antibody chromatographic film, water suction bar, the bottom plate assembled, the bottom plate of assembling is cut, food pathogenic infection detector bar is obtained.
The detection means of the information-based food pathogenic infection of rapid qualitative of the present invention is to be based on following principle：
Sample is the extract of excrement or excrement.Sample application zone (the 1st area) is added drop-wise to, sample need not be handled, and borrowed or do not borrowed
Help developping solution to go upward to impurity in filtering area (the 2nd area), sample and be stranded in filtering area, the liquid dissolving solid phase labelling antibody of filtration
The labelled antibody in area (the 4th area) enters reaction zone (the 5th, 6th area).Such as there is antigen in sample, then it is " anti-with labelled antibody formation
Original-labelled antibody " compound.The compound is up on chromatography strip by capillary theory, to the 5th area's detection zone, with T areas
Another specific mark antibody formation " labelled antibody-antigen-solid phase labelling antibody " composite precipitation line of one solidus.This sinks
The color of shallow lake line will be in direct ratio with concentration in intensity antigen blood sample.Double antibody sandwich method principle-in T detection zones formation " solid phase
It is coated with labelled antibody-determined antigen-labelled antibody " immune complex, its color intensity is with labelled antigen concentration to be measured into just
Than.Such as 10-15 minutes particular detection time, chromatography detector bar (box) is detected by an unaided eye and records result or quantitative analysis instrument
Middle reading result.
Preferably, it is described quantitative in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention
Detector is handheld portable detector.
Preferably, it is described hand-held in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention
Portable detector is made up of the readout instrument based on smart mobile phone, inspection software, cloud service.
Preferably, in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention, the transmission
The transmission means of device includes the one or more of following methods：
By the cloud service, through wechat downloading data；
By the output software of the instrument, through USB output datas；
Printed out from wechat or bluetooth printing machine；
Sent by the readout instrument by lettergram mode.
Preferably, it is described hand-held in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention
Stored in the inspection software of portable detector testing sample food pathogenic infection concentration and detector color intensity it
Between linear relationship.
Preferably, in the detection means of the information-based food pathogenic infection of rapid qualitative of the present invention, the food
Pathogenic infection includes rotavirus, norovirus, adenovirus, helicobacter pylori, large intestine toxic bacillus, salmonella, Li Shi
The one or more of bacillus.
Another aspect of the present invention is to provide a kind of detection method of the information-based food pathogenic infection of rapid qualitative, including
Following steps：
1) testing sample is added dropwise to the sample uptake zone of above-mentioned food pathogenic infection detector bar；
2) the sample chromatography enters above-mentioned filtering area, chromatographic zone；
If 3) there is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line in chromatographic zone, illustrate detection knot
Fruit is the positive, if there is not the composite precipitation line, illustrates to be detected as feminine gender；When the sample enters check plot,
Control line should occur, when such as there is no control line, detection failure；
4) " labelled antibody-antigen-solid-phase coating antibody " the composite precipitation line is carried out using above-mentioned instrument
Color intensity is determined, and obtains antigen concentration in sample, is transmitted to the terminal count and filled by preceding claim transmitting device
Put.
Compared with prior art, the present invention has advantages below to the present invention：
1. it is specific high：Required pairing labelled antibody acts only on high specific detection food pathogenic infection.
2. sensitivity is high：Required pairing labelled antibody is with high-sensitivity detection food pathogenic infection.With enzyme linked immunosorbent assay, luminescence method
It is close, 0.1ng/ml detection level can be reached.
3. detection box detection food pathogenic infection trial zone (T) shade or p-wire that required pairing antibody is fabricated to
Intensity and food pathogenic infection concentration are linear, can by being stored in the detector and network transmission device of cloud service
Easily to obtain quantitative testing result.
4. can a step quick detection, sample need to be only added dropwise in the detection box that required pairing labelled antibody is fabricated to, at 10 points
Clock just can measure result.
5. testing result quantitative information, by the way that food pathogenic infection detector bar is inserted into detecting instrument, at once into number
Word program display, and can be by relevant information by computer or any network equipment, or smart mobile phone, storage sends to be measured
Person, such as operating personnel, doctor, or it is sent to inspection center.
Brief description of the drawings
Fig. 1 is the information-based food pathogenic infection detector bar structural representation of rapid qualitative of the present invention；
Fig. 2 is the testing result schematic diagram of individual event food pathogenic infection detector bar in the embodiment of the present invention；
Fig. 3 is the testing result schematic diagram of individual event food pathogenic infection detection box in the embodiment of the present invention；
Fig. 4 is the testing result schematic diagram of double item food pathogenic infections detection boxes in the embodiment of the present invention；
Fig. 5 illustrates for the outward appearance of handheld portable detector in the food pathogenic infection detection means in the embodiment of the present invention
Figure；
Fig. 6 is signal schematic diagram in smart mobile phone position in the food pathogenic infection detection means in the embodiment of the present invention；
Fig. 7 illustrates for quantitative information result output display in the food pathogenic infection detection means in the embodiment of the present invention
Figure.
Embodiment
An embodiment provides a kind of detection means of the information-based food pathogenic infection of rapid qualitative, bag
Include
Food pathogenic infection detector bar；
Instrument, is connected by the neck and the food pathogenic infection detector bar that are set on the instrument
Connect, obtain the quantitative testing result of the food pathogenic infection detector bar；
Transmitting device, the quantitative testing result for transmitting the instrument detection；
And terminal count device, the quantitative testing result for reading the transmitting device transmission.
In one embodiment of the invention, the food pathogenic infection detector bar has following structure, as shown in figure 1,：
1st area is sample uptake zone (sample application zone),
2nd area is filtering area,
3rd area is chromatographic zone (nitrocellulose chromatographic film),
4th area immobilized label antibody district A,
5th area is detection zone T, including labelled antibody spraying area band,
6th area is check plot C,
7th area is suction zones,
Wherein, the chromatographic zone includes immobilized label antibody district, detection zone and check plot.
In one embodiment of the invention, the sample uptake zone of food pathogenic infection detector bar of the present invention
(bar), filtering area (bar), chromatographic zone (bar) and suction zones (bar) are arranged on bottom plate；The present invention is examined to the food pathogenic infection
The assembling of survey article is preferably first from bottom to top to paste sample uptake zone (the 1st area), filtering area (the 2nd area), chromatography successively in bottom plate
Area (the 3rd area), suction zones (the 7th area), paste make the overlapping 1~1.5mm in each area on viscose glue bottom plate, during stickup.
The present invention does not have special limitation to the material, size and source of the bottom plate, ripe using those skilled in the art
The bottom plate being used in immunoassay strip known.In the present invention, the material of the bottom plate is preferably plastics or film condensation material.
After the bottom plate assembled, the bottom plate of assembling is cut, immunity test strip is obtained.The present invention does not have to the method for the slitting
Have specifically limited, using slitting method well-known to those skilled in the art.In the present invention, the width of the slitting is preferred
For 3.0~5.0mm, preferably 4mm.
The immunoassay strip that the present invention is provided includes the sample uptake zone being arranged on the bottom plate.In the present invention, institute
State the material preferably glass fibre of sample uptake zone.The present invention does not have any limitation, ability to the source of the glass fibre
Glass fibre known to field technique personnel.The present invention is pasted to the sample uptake zone in the lower end of bottom plate, will be described
The lower edge of sample uptake zone is concordant with bottom plate lower edge.
In the present invention, the sample that the sample uptake zone absorbs is excrement or the extract of excrement.The source of the excrement
Do not limit, human or animal's excrement.
In one embodiment of the invention, the solid phase labelling antibody district is arranged on the bottom plate, positioned at filtering area
Top, and the solid phase labelling antibody district 1~2mm overlapping with filtering area.In one embodiment of the invention, it is described solid
The material in phase labelled antibody area is preferably glass fibre.
In one embodiment of the invention, the reaction zone is close to the solid phase labelling antibody district, positioned at the solid phase
The top in labelled antibody area, and 1~2mm of the reaction zone and the solid phase labelling antibody area overlapping.At one of the present invention
In embodiment, the material of the reaction zone is preferably nitrocellulose filter.
In one embodiment of the invention, the reaction zone is provided with detection line T and nature controlling line C.Described detection line T
Below nature controlling line C, the detection line T is close to solid phase labelling antibody district, and the nature controlling line C is close to suction zones.
In one embodiment of the invention there is provided the preparation method of the detector bar of food pathogenic infection, including it is following
Step：
Labelled antibody is dissolved in buffer solution, the labelled antibody diluted, the labelled antibody of the dilution is sprayed
On glass fibre membrane, solid phase labelling antibody membrane is obtained after drying, solid phase labelling antibody district is used as；
The reaction zone that coated antibody solution is lined into nitrocellulose film makes detection line, and rabbit anti-mouse igg solution is rule
Nature controlling line is made in the reaction zone of nitrocellulose film, obtains being coated with labelled antibody chromatographic film after drying, is used as reaction zone；
Sample absorbing strip, filtering rod, the solid phase labelling antibody membrane, the coating mark are from bottom to top pasted successively in bottom plate
Remember after antibody chromatographic film, water suction bar, the bottom plate assembled, the bottom plate of assembling is cut, food pathogenic infection detector bar is obtained.
In the present invention, labelled antibody is added into colloid gold label solution, after stirring 10~15 minutes, it is 1% to add concentration
Bovine serum albumin liquid, the Bovine serum albumin liquid and colloid gold label liquor capacity ratio are 1：7~15, through at a high speed
After (12000PM) is centrifuged ten minutes, colored precipitate thing is obtained, the colored precipitate thing is labelled antibody.Serum solution in extraction, sinks
Starch is dissolved in specific buffer solution.
The present invention is not particularly limited to the source of the colloid gold label solution, using well known to those skilled in the art
Colloid gold label solution.
In the present invention, the centrifugal rotational speed is preferably 12000vpm, and the temperature of the centrifugation is 4 DEG C.
In the present invention, the method for the labelled antibody is not particularly limited, dilution side well known to those skilled in the art
Method.
Obtain after labelled antibody, mark is coated on glass fibre membrane.The coating is preferably immersion, hand coatings or instrument
Device is sprayed, using the method for immersion in the embodiment of the present invention.Labelled antibody is then by the tunica fibrosa after coating in vacuum desiccator
Under, it is dried in vacuo, obtains solid phase labelling antibody district.In the present invention, the temperature of the drying is preferably 35~38 DEG C, 8
Solid phase labelling antibody membrane is made in or so hour.
The preparation of coated antibody：
The vacuum drying of labelled antibody solution is lined into making detection line T lines on nitrocellulose film (the 5th area) reaction zone, will
Rabbit anti-mouse igg solution, which is lined, makes nature controlling line C line (the 6th area), the check plot rule on reaction zone.The present invention is to described
The mode of line is not restricted, using labelled antibody spraying method well known to those skilled in the art.The present invention is implemented
Line is rule using lining instrument in example, model Biojet, the Biodot CA, USA of the lining instrument.In the present invention one
In individual embodiment, the concentration of the coating buffer is preferably 2mg/ml；The coating buffer quantity for spray is preferably per 30cm nitrocelluloses
Film is sprayed 80ul/ seconds.
After line, nitrocellulose film is placed under 37 DEG C of environment and dried 25 minutes by the present invention, obtains coating labelled antibody layer
Analyse film.The present invention is not particularly limited to the method for the drying, is using furnace drying method well-known to those skilled in the art
Can.In an embodiment of the invention, sample absorbing strip, filtering rod, mark chromatography strip and water suction bar are pasted onto viscose glue bottom plate
On, the bottom plate assembled.In an embodiment of the invention, it is preferably first from bottom to top to be glued successively in bottom plate to the assembling
Note sample uptake zone (the 1st area), filtering area (the 2nd area), chromatographic zone (the 3rd area), suction zones (the 7th area), are pasted on viscose board,
Make the overlapping 1~2mm in each area during stickup.
After the bottom plate assembled, the bottom plate of assembling is cut, the detector bar of food pathogenic infection is obtained.The present invention
Method to the slitting is not particularly limited, using slitting method well-known to those skilled in the art.It is of the invention real
Apply slitting in example to cut using cutting machine, the cutting machine model K:nematic2360CA,USA.In one implementation of the present invention
In example, the width of the slitting is preferably 3.0~5.0mm, preferably 4mm.
In an embodiment of the invention, described antibody labeling particle can be collaurum, and magnetic particle, carbon particle shines grain,
Glue gold body grain, silver granuel, colored latex grain, fluorescence, textile dyestuff, enzyme, quantum dot, liposome, other particles etc..In the present invention one
In individual embodiment, the described preferred collaurum of antibody labeling particle.
In an embodiment of the invention, described food pathogenic infection is preferably rotavirus, norovirus, adenopathy
Poison, helicobacter pylori, large intestine toxic bacillus, salmonella or Li bacillus.
In embodiments of the invention, the antibody of the corresponding antibody of the food pathogenic infection and labelled antibody, antigen used
Standard items are inactivation, and purified antigenic source is all from JAJ international, Inc.San Diego, CA, USA.
The coated antibody of food pathogenic infection and the product type of labelled antibody described in the embodiment of the present invention see the table below 1.
The food pathogenic infection of table 1 matches labelled antibody product type title
The detection means of the information-based food pathogenic infection of rapid qualitative of the present invention is to be based on following principle：
Sample is the extract of excrement or excrement.Sample application zone (the 1st area) is added drop-wise to, sample need not be handled, and borrowed or do not borrowed
Help developping solution to go upward to impurity in filtering area (the 2nd area), sample and be stranded in filtering area, the liquid dissolving solid phase labelling antibody of filtration
The labelled antibody in area (the 4th area) enters reaction zone (the 5th, 6th area).Such as there is antigen in sample, then it is " anti-with labelled antibody formation
Original-labelled antibody " compound.The compound is up on chromatography strip by capillary theory, to the 5th area's detection zone, with T areas
Another specific mark antibody formation " labelled antibody-antigen-solid phase labelling antibody " composite precipitation line of one solidus.This sinks
The color of shallow lake line will be in direct ratio with concentration in intensity antigen blood sample.Double antibody sandwich method principle-in T detection zones formation " solid phase
It is coated with labelled antibody-determined antigen-labelled antibody " immune complex, its color intensity is with labelled antigen concentration to be measured into just
Than.Such as 10-15 minutes particular detection time, chromatography detector bar (box) is detected by an unaided eye and records result or quantitative analysis instrument
Middle reading result.
Preferably, in one embodiment of the invention, the instrument is handheld portable detector.
Preferably, in one embodiment of the invention, the handheld portable detector is based on smart mobile phone
Readout instrument, inspection software, cloud service composition.
Preferably, in one embodiment of the invention, the transmission means of the transmitting device includes the one of following methods
Plant or several：
By the cloud service, through wechat downloading data；
By the output software of the instrument, through USB output datas；
Printed out from wechat or bluetooth printing machine；
Sent by the readout instrument by lettergram mode.
In an embodiment of the present invention, quantitative information readout instrument used is QikTech-4000 (JAJ
International, Inc.San Diego, CA, USA), it is a kind of image immunoassays complete equipment, by " in smart mobile phone
Based on readout instrument；Inspection software；Cloud service " is constituted.
QikTech-4000 is handheld portable detector, its outward appearance as shown in figure 5, will detect box (detector bar when using
Outer layer sets the detection box of plastic housing) insertion detector, based on intelligent quick diagnosis test (RDT) reader, carry out image and adopt
Collection, processing and analysis.
Detect the color intensity relation proportional to test substance of detection line in box.This color or luminous intensity are color
Intensity is marked such as gold, or is fluorescence intensity, or is luminous intensity.Color intensity readout instrument can be read to the first image of testing result
As a result, then Digitization Software is analyzed.Most a whole set of result is inputted to high in the clouds at last.Once will detection box insertion readout instrument, intelligence
The open lighting device of mobile phone hardware communication, this lighting device (smart mobile phone and its connection with other parts below detection box
Relation is as shown in Figure 6), this narrow LED light, through detection box, detects detection line (T) color intensity, passes through intelligence
Mobile phone enters digital shadow record program.Intensity to be measured is made comparisons with the standard curve (see embodiment) stored in software, is directly shown
The concentration of test substance is shown：Units per ml, the testing result of quantitative information is as shown in Figure 7.
In the present invention, the food pathogenic infection of testing sample has been stored in the inspection software of the handheld portable detector
Linear relationship between concentration and the color intensity of detector bar.
In the present invention, the food pathogenic infection includes rotavirus, norovirus, adenovirus, helicobacter pylori, big
Intestines toxic bacillus, salmonella, the one or more of Li bacillus.
The invention provides a kind of detection method of the information-based food pathogenic infection of rapid qualitative, comprise the following steps：
1) testing sample is added dropwise to the sample uptake zone of above-mentioned food pathogenic infection detector bar；
2) sample by or not by developping solution, into above-mentioned filtering area, chromatographic zone；
If 3) as shown in Fig. 2 there is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line in chromatographic zone,
It is the positive to illustrate testing result, if there is not the composite precipitation line, illustrates to be detected as feminine gender；When the sample enters
During check plot, control line (shown in Fig. 2) should occur, when such as there is no control line, detection failure；
4) " labelled antibody-antigen-solid-phase coating antibody " the composite precipitation line is carried out using above-mentioned instrument
Color intensity is determined, and is obtained antigen concentration in sample, is transmitted by above-mentioned network transmission device to the terminal count device.
In the present invention, the food pathogenic infection detector bar outer layer can be provided with plastic casing, as shown in figure 3, being list
A kind of result schematic diagram of item (only having food pathogenic infection in sample) detection box (detector bar), wherein, occur " labelled antibody-
Antigen-solid-phase coating antibody " composite precipitation line T, then it is the positive to illustrate testing result, if there is not the composite precipitation line
(T), then explanation is detected as feminine gender；When the sample enters check plot, control line C (shown in Fig. 3) should occur, not compare such as
During line, then detection failure.
In the present invention, the food pathogenic infection detector bar can test two or more food pathogenic infection,
As shown in figure 4, be the result schematic diagram of double items (only having two kinds of food pathogenic infections in sample) detection box (detector bar), wherein,
There is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line T1, T2, then illustrate testing result for two kinds of food senses
Former (such as rotavirus, norovirus) is caught an illness for positive findings, if there is not the composite precipitation line (T), is illustrated
It is detected as feminine gender；When the sample enters check plot, control line C (shown in Fig. 3) should occur, when such as there is no control line, then examine
Dendrometry loses.
Below in conjunction with the embodiment in the present invention, the technical scheme in the present invention is clearly and completely described.It is aobvious
So, described embodiment is only a part of embodiment of the invention, rather than whole embodiments.Based on the reality in the present invention
Example is applied, the every other embodiment that those of ordinary skill in the art are obtained under the premise of creative work is not made all belongs to
In the scope of protection of the invention.
The configuration of the rotavirus RoTa Virus detection means of embodiment 1
1) setting of rotavirus RoTa Virus detector bars
By product type for V-0812 rotavirus RoTa Virus labelled antibodies (JAJ international,
Inc.San Diego, CA, USA) colloid gold label solution is added, after stirring 10~15 minutes, add the calf that concentration is 1%
Haemocyanin liquid, the Bovine serum albumin liquid is 1 with colloid gold label liquor capacity ratio：10, through 4 DEG C of (12000PM) at a high speed
After centrifugation 10 minutes, colored precipitate thing is obtained, the colored precipitate thing is labelled antibody.Serum solution in extraction, sediment dissolving
In phosphate buffer.Labelled antibody mark after dilution is immersed on glass fibre membrane, by the tunica fibrosa after immersion true
Under empty drier, 37 DEG C are dried in vacuo, and solid phase labelling antibody membrane are made within 8 hours or so.
Concentration containing coated antibody V-0428 in coating buffer is 2mg/ml, and coating buffer is lined into nitrocellulose membrane
On, make detection zone (the 5th area).Coating buffer quantity for spray is to be sprayed 80ul/ seconds per 30cm nitrocelluloses film.By 3 mg/mls
Rabbit anti-mouse igg solution (product type #125-3401) lined according to foregoing spraying rate nature controlling line C line made on reaction zone
(the 6th area), the check plot rule.Line is rule using lining instrument, the model Biojet of the lining instrument,
Biodot CA,USA.After line, nitrocellulose film is placed under 37 DEG C of environment and dried 25 minutes by the present invention, obtains labelled antibody
Chromatographic film.
Sample absorbing strip, filtering rod, solid phase labelling antibody membrane, labelled antibody chromatographic film and water suction bar are pasted onto viscose glue bottom
On plate, the bottom plate assembled.Assemble method is first in bottom plate from bottom to top paste sample uptake zone (the 1st area), filtering successively
Area (the 2nd area), chromatographic zone (the 3rd area), suction zones (the 7th area), paste make the overlapping of each area on viscose glue bottom plate, during stickup
1.5mm.After the bottom plate assembled, the bottom plate of assembling is cut, slitting is using cutting machine slitting, the cutting machine model
For K:nematic2360CA,USA.The width of the slitting is 4mm.Obtain rotavirus RoTa Virus detector bar.
2) in rotavirus RoTa Virus detection means instrument setting：
By rotavirus RoTa Virus standard samples by following concentration dilution into phosphate buffer, 10,20,50,
100、500(ng/ml).Above-mentioned 100ul microlitres (1 drop) is added in detector bar sample application zone, in detection box being placed in after 10 minutes
Instrument reading, reading result summary sheet be stored in detector QikTech-4000 (JAJ international,
Inc.San Diego, CA, USA) carry it is standby in software, instrument by table 2 below obtain precipitation line color intensity (OD values) and
Rotavirus RoTa Virus antigen concentration distribution curves in excrement, and be stored in cloud server.
The color intensity testing result of the different rotavirus RoTa Virus antigen concentrations of table 2
| Rotavirus RoTa Virus (ng/ml) | Color intensity |
| 10 | 0.98 |
| 20 | 1.24 |
| 50 | 2.31 |
| 100 | 3.02 |
| 500 | 5.11 |
3) in rotavirus RoTa Virus detection means network transmission device setting
Above-mentioned data are by the cloud service, and data receiver (such as inspection center doctor) is through wechat downloading data.
The configuration of the norovirus Nono Virus detection means of embodiment 2
1) the setting be the same as Example 1 of norovirus Nono Virus detector bars, only replaces coated antibody and labelled antibody
For V-1246 and V-1262.
2) in norovirus Nono Virus detection means instrument setting be the same as Example 1.Add above-mentioned 100ul
Microlitre (1 drop) in detector bar sample application zone, in detection box is placed in into instrument reading, reading result summary sheet after 10 minutes
Be stored in detector QikTech-4000 (JAJ international, Inc.San Diego, CA, USA) carry it is standby in software
With instrument obtains the color intensity (OD values) and rotavirus RoTa Virus antigen concentrations in excrement of precipitation line by table 3 below
Distribution curve, and be stored in cloud server.
The color intensity testing result of the different norovirus Nono Virus antigen concentrations of table 3
| Norovirus Nono Virus (ng/ml) | Color intensity |
| 10 | 0.47 |
| 20 | 0.72 |
| 50 | 1.31 |
| 100 | 2.01 |
| 500 | 4.02 |
3) in norovirus Nono Virus detection means network transmission device setting
Above-mentioned data are via detector QikTech-4000 output softwares, through USB output datas.
The configuration of the adenovirus Adeno Virus detection means of embodiment 3
1) the setting be the same as Example 1 of adenovirus Adeno Virus detector bars, only replaces with coated antibody with labelled antibody
V-2082 and V-2124.
2) in adenovirus Adeno Virus detection means instrument setting be the same as Example 1.Add above-mentioned 100ul
Microlitre (1 drop) in detector bar sample application zone, in detection box is placed in into instrument reading, reading result summary sheet after 10 minutes
Be stored in detector QikTech-4000 (JAJ international, Inc.San Diego, CA, USA) carry it is standby in software
With instrument obtains the color intensity (OD values) and rotavirus RoTa Virus antigen concentrations in excrement of precipitation line by table 3 below
Distribution curve, and be stored in cloud server.
The color intensity testing result of the different adenovirus Adeno Virus antigen concentrations of table 4
| Adenovirus Adeno Virus (ng/ml) | Color intensity |
| 10 | 0.97 |
| 20 | 1.42 |
| 50 | 2.22 |
| 100 | 4.01 |
| 500 | 6.12 |
3) in adenovirus Adeno Virus detection means network transmission device setting
Above-mentioned data are printed out from wechat or bluetooth printing machine.
The configuration of the helicobacter pylori Helicobacter Pylori detection means of embodiment 4
1) the setting be the same as Example 1 of helicobacter pylori Helicobacter Pylori detector bars, only by coated antibody with
Labelled antibody replaces with B-1432 and B-1042.
2) in helicobacter pylori Helicobacter Pylori detection means instrument setting be the same as Example 1.
Above-mentioned 100ul microlitres (1 drop) is added in detector bar sample application zone, in detection box is placed in into instrument reading after 10 minutes,
Reading result summary sheet is stored in detector QikTech-4000 (JAJ international, Inc.SanDiego, CA, USA)
Carry standby in software, color intensity (OD values) and rotavirus RoTa in excrement that instrument passes through table 3 below acquisition precipitation line
Virus antigen concentration distribution curves, and be stored in cloud server.
The color intensity testing result of the different helicobacter pylori Helicobacter Pylori antigen concentrations of table 5
3) in helicobacter pylori Helicobacter Pylori detection means network transmission device setting
Above-mentioned data are via detector QikTech-4000 output softwares, through USB output datas.
The large intestine toxic bacillus E Coli-0157 of embodiment 5:The configuration of H7 detection means
1) large intestine toxic bacillus E Coli-0157:The setting be the same as Example 1 of H7 detector bars, only by coated antibody and mark
Antibody replaces with B-3402 and B-3104.
2) large intestine toxic bacillus E Coli-0157:The setting be the same as Example 1 of instrument in H7 detection means.Add
Above-mentioned 100ul microlitres (1 drop) in detector bar sample application zone, in detection box is placed in into instrument reading, reading after 10 minutes
As a result summary sheet is stored in detector QikTech-4000 (JAJ international, Inc.SanDiego, CA, USA) and carried
Standby in software, instrument obtains the color intensity (OD values) and rotavirus RoTa Virus in excrement of precipitation line by table 3 below
Antigen concentration distribution curve, and be stored in cloud server.
The difference large intestine toxic bacillus E of table 6 Coli-0157:The color intensity testing result of H7 antigen concentrations
3) large intestine toxic bacillus E Coli-0157:The setting of network transmission device in H7 detection means
Above-mentioned data are sent from the readout instrument of the detector by lettergram mode.
The configuration of the salmonella Salmonella Typhi detection means of embodiment 6
1) the setting be the same as Example 1 of salmonella Salmonella Typhi detector bars, it is only that coated antibody is anti-with mark
Body replaces with B-4122 and B-4212.
2) in salmonella Salmonella Typhi detection means instrument setting be the same as Example 1.In addition
100ul microlitres (1 drop) is stated in detector bar sample application zone, in detection box is placed in into instrument reading, reading knot after 10 minutes
Fruit summary sheet be stored in detector QikTech-4000 (JAJ international, Inc.SanDiego, CA, USA) carry it is soft
Standby in part, instrument obtains the color intensity (OD values) of precipitation line by table 3 below and rotavirus RoTa Virus in excrement resist
Original content distribution curve, and be stored in cloud server.
The color intensity testing result of the different salmonella Salmonella Typhi antigen concentrations of table 7
3) in salmonella Salmonella Typhi detection means network transmission device setting
Via detector QikTech-4000 output softwares, through USB output datas.
The configuration of the Li bacillus Listeria detection means of embodiment 7
1) the setting be the same as Example 1 of Li bacillus Listeria detector bars, only replaces with coated antibody with labelled antibody
B-5032 and B-5144.
2) in Li bacillus Listeria detection means instrument setting be the same as Example 1.Add above-mentioned 100ul micro-
(1 drop) is risen in detector bar sample application zone, in detection box is placed in into instrument reading after 10 minutes, reading result summary sheet is stored up
Be stored in detector QikTech-4000 (JAJ international, Inc.San Diego, CA, USA) carry it is standby in software,
Instrument obtains the color intensity (OD values) of precipitation line by table 3 below and rotavirus RoTa Virus antigen concentrations in excrement are distributed
Curve, and be stored in cloud server.
The color intensity testing result of the different Li bacillus Listeria antigen concentrations of table 8
| Li bacillus Listeria (ng/ml) | Color intensity |
| 10 | 0.54 |
| 20 | 0.98 |
| 50 | 1.38 |
| 100 | 2.06 |
| 500 | 3.42 |
3) in Li bacillus Listeria detection means network transmission device setting
Above-mentioned data are sent from the readout instrument of the detector by lettergram mode.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of detection means of the information-based food pathogenic infection of rapid qualitative, including
Food pathogenic infection detector bar；
Instrument, is connected with the food pathogenic infection detector bar by the neck set on the instrument, obtained
To the quantitative testing result of the food pathogenic infection detector bar；
Transmitting device, the quantitative testing result for transmitting the instrument detection；
And terminal count device, the quantitative testing result for reading the transmitting device transmission.
2. the detection means of the information-based food pathogenic infection of rapid qualitative according to claim 1, it is characterised in that institute
Stating food pathogenic infection detector bar includes sample uptake zone, filtering area, chromatographic zone and suction zones.
3. the detection means of the information-based food pathogenic infection of rapid qualitative according to claim 2, it is characterised in that institute
Stating chromatographic zone includes immobilized label antibody district and reaction zone, and the reaction zone includes detection zone and check plot.
4. the detection means according to claims 1 to 3 any one, it is characterised in that the food pathogenic infection detection
The preparation method of bar, comprises the following steps：
Labelled antibody is dissolved in buffer solution, the labelled antibody of the dilution is sprayed on glass by the labelled antibody diluted
On glass tunica fibrosa, solid phase labelling antibody membrane is obtained after drying；
The reaction zone that coated antibody solution is lined into nitrocellulose film makes detection line, and rabbit anti-mouse igg solution is lined into nitre
The reaction zone of base tunica fibrosa makes nature controlling line, obtains being coated with labelled antibody chromatographic film after drying；
In bottom plate, from bottom to top paste sample absorbing strip, filtering rod, the solid phase labelling antibody membrane, coating mark are anti-successively
After body chromatographic film, water suction bar, the bottom plate assembled, the bottom plate of assembling is cut, food pathogenic infection detector bar is obtained.
5. the detection means of the information-based food pathogenic infection of rapid qualitative according to claim 1, it is characterised in that institute
Instrument is stated for handheld portable detector.
6. the detection means of the information-based food pathogenic infection of rapid qualitative according to claim 5, it is characterised in that institute
Handheld portable detector is stated to be made up of the readout instrument based on smart mobile phone, inspection software, cloud service.
7. the detection means of the information-based food pathogenic infection of rapid qualitative according to claim 6, it is characterised in that institute
Stating the transmission means of transmitting device includes the one or more of following methods：
By the cloud service, through wechat downloading data；
By the output software of the instrument, through USB output datas；
Printed out from wechat or bluetooth printing machine；
Sent by the readout instrument by lettergram mode.
8. the detection means of the information-based food pathogenic infection of rapid qualitative according to claim 6, it is characterised in that institute
The food pathogenic infection concentration and the food that testing sample has been stored in the inspection software for stating handheld portable detector infect
Linear relationship between the color intensity of Pathogen test instrument.
9. the detection means of the information-based food pathogenic infection of rapid qualitative according to claims 1 to 8, its feature exists
In the food pathogenic infection includes rotavirus, norovirus, adenovirus, helicobacter pylori, large intestine toxic bacillus, sand
Door Salmonella and the one or more of Li bacillus.
10. a kind of detection method of the information-based food pathogenic infection of rapid qualitative, comprises the following steps：
1) sample for the food pathogenic infection detector bar being added dropwise to testing sample described in claim 2~9 any one absorbs
Area；
2) sample chromatography right of access profit requires filtering area described in 2~9 any one, chromatographic zone；
If 3) there is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line in chromatographic zone, illustrate that testing result is
The positive, if there is not the composite precipitation line, illustrates to be detected as feminine gender；When the sample enters check plot, it should go out
Existing control line, when such as there is no control line, detection failure；
4) application claim 1~9 any one described in instrument to it is described " labelled antibody-antigen-solid-phase coating resist
Body " composite precipitation line carries out color intensity measure, antigen concentration in sample is obtained, by described in claim 1~9 any one
Transmitting device transmit to the terminal count device.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060222567A1 (en) * | 2005-04-01 | 2006-10-05 | Hafellner | Body fluid testing component for simultaneous analyte detection |
| CN105164514A (en) * | 2013-01-21 | 2015-12-16 | 康奈尔大学 | Smartphone-based apparatus and method for obtaining repeatable, quantitative colorimetric measurement |
| CN105717294A (en) * | 2014-12-16 | 2016-06-29 | 生物梅里埃公司 | Method and device for determining the presence of a micro-organism in stools with activated carbon pretreatment |
-
2017
- 2017-07-05 CN CN201710542832.XA patent/CN107192824A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060222567A1 (en) * | 2005-04-01 | 2006-10-05 | Hafellner | Body fluid testing component for simultaneous analyte detection |
| CN105164514A (en) * | 2013-01-21 | 2015-12-16 | 康奈尔大学 | Smartphone-based apparatus and method for obtaining repeatable, quantitative colorimetric measurement |
| CN105717294A (en) * | 2014-12-16 | 2016-06-29 | 生物梅里埃公司 | Method and device for determining the presence of a micro-organism in stools with activated carbon pretreatment |
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