CN105606800B - A kind of broad-spectrum pesticide detects gold test strip and method - Google Patents
A kind of broad-spectrum pesticide detects gold test strip and method Download PDFInfo
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- CN105606800B CN105606800B CN201610054574.6A CN201610054574A CN105606800B CN 105606800 B CN105606800 B CN 105606800B CN 201610054574 A CN201610054574 A CN 201610054574A CN 105606800 B CN105606800 B CN 105606800B
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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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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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
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
- G01N33/521—Single-layer analytical elements
- G01N33/523—Single-layer analytical elements the element being adapted for a specific analyte
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2430/00—Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes
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Abstract
The invention discloses a kind of broad-spectrum pesticide detection gold test strip and method, the detection line that test paper has detection line, control line and indicator, test paper is provided with cholinesterase, and control line is provided with the capture factor；Capture mark A and B are connected with indicator, wherein capture mark A can be captured by cholinesterase, capture mark B can be by the capture factor capture on control line.Various Organophosphorus and carbamate pesticides class agricultural chemicals are respectively provided with preferable detectability by Pesticides Testing test paper of the invention, and testing result is fast, are highly suitable for the quick Detecting Pesticide in scene of vegetables and fruits.Pesticides Testing test paper of the invention, preparation process is simple, testing result reliability, it is possible to achieve the Detecting Pesticide of low cost.
Description
Technical field
The present invention relates to a kind of Detecting Pesticide test paper and method, more particularly to a kind of broad-spectrum pesticide detection gold test strip
And method.
Background technology
Existing conventional agricultural chemicals has the major class of Organophosphorus and carbamate pesticides two, specific all kinds of up to 40 kinds, conventional
There are nearly 20 kinds.It is a kind of method for effectively reducing crop disease and insect using agricultural chemicals, but irrational use agricultural chemicals can cause agriculture
Medicine is remained, and threatens the food security of people.
With the food-safe pay attention to day by day of people, increasing people begins to focus on residues of pesticides.Traditional agricultural chemicals
Method for detecting residue has chromatography, and the detection accuracy of this method is high, but time-consuming, high to operation and the requirement of instrument,
The need for field quick detection cannot being met.
Immunologic detection method based on antibody-antigene mainly uses antibody to the specific recognition of antigen, and ties therewith
Close, with high precision, it is sensitive, special the characteristics of.It is simultaneously low to the requirement for operating, more can quickly provide qualitative detection
As a result.Agricultural chemicals quick detection reagent based on the exploitation of this principle has many kinds.But the species of agricultural chemicals is various, and immune detection
The high specific of method causes it can only to detect a certain specific agricultural chemicals.If realizing complete detection, need to use
More than 20 kinds of agricultural chemicals immunologic function test reagent, this will greatly increase the cost of agricultural chemicals quick detection, cause comprehensive inspection of agricultural chemicals
Survey it is practically impossible to realize.Substantial amounts of agricultural chemicals cannot be detected, and there are potential risks.
The content of the invention
It is an object of the invention to provide a kind of Pesticides Testing gold test strip of wide spectrum, the test paper can be gone out several with quick detection
Whole Organophosphorus and carbamate pesticides class agricultural chemicals.
The technical solution used in the present invention is：
A kind of broad-spectrum pesticide Test paper, test paper has detection line, control line and indicator,
The detection line of test paper is provided with cholinesterase, and control line is provided with the capture factor；
Capture mark A and B are connected with indicator, wherein capture mark A can be captured by cholinesterase, and capture mark B can
By the capture factor capture on control line.
Preferably, indicator is physical tracer.
Preferably, capture mark A and organophosphor or carbamate chemicals for agriculture competitiveness are combined with cholinesterase.
Preferably, capture mark A is organophosphor or carbamate chemicals for agriculture, organophosphor or carbamate chemicals for agriculture
Characteristic residue, or organophosphor or carbamate chemicals for agriculture conjugate.
Preferably, capture mark B be albumen family macromolecule, polypeptide polymer, degradable high molecular polymer, it is corresponding
The capture factor is its antibody；
Or capture mark B is oligonucleotide sequence, the corresponding capture factor is its complementary series；
Or capture mark B and its capture factor constitute affine system pair.
Preferably, organophosphor or carbamate chemicals for agriculture are selected from parathion, fenifrothion, parathion-methyl, Rogor, water
Amine sulphur phosphorus, malathion, chlopyrifos and its conjugate.Organophosphor or carbamate chemicals for agriculture characteristic residue are selected from double sulphur
Base.
Preferably, capture mark B and its capture factor constitute affine system to being biotin-avidin system.
Preferably, indicator is collaurum.
A kind of agricultural chemicals method for quick, including sample is processed, sampling afterwards is added dropwise and detects examination in broad-spectrum pesticide
On paper, in the case of the instruction according to instruction, testing result is determined, wherein, broad-spectrum pesticide Test paper is as described above.
The beneficial effects of the invention are as follows：
Various Organophosphorus and carbamate pesticides class agricultural chemicals are respectively provided with preferably detection energy by Pesticides Testing test paper of the invention
Power, testing result is fast, is highly suitable for the quick Detecting Pesticide in scene of vegetables and fruits.
Pesticides Testing test paper of the invention, preparation process is simple, testing result reliability, it is possible to achieve the agricultural chemicals of low cost is residual
Stay detection.
Specific embodiment
A kind of broad-spectrum pesticide Test paper, test paper has detection line, control line and indicator,
The detection line of test paper is provided with cholinesterase, and control line is provided with the capture factor；
Capture mark A and B are connected with indicator, wherein capture mark A can be captured by cholinesterase, and capture mark B can
By the capture factor capture on control line.
Capture mark A and B can distinguish direct or indirect connection on indicator.Or capture mark A is connected to capture
On mark B, then it is connected with indicator again.In general, when capture mark A and B are small molecule, A and B are equal for capture mark
Independent connection can avoid interfering on indicator, obtain more preferable testing result.And work as capture mark B for macromolecular
When, capture mark A can be first connected on capture mark B, and then capture mark B is connected with indicator again.
The Cleaning Principle of test paper of the present invention is as follows：
Organophosphorus and carbamate pesticides class agricultural chemicals can be combined with cholinesterase and be suppressed its activity, organophosphor and amino first
Capture mark A competitiveness on esters of gallic acid agricultural chemicals, indicator is combined with cholinesterase.Therefore, when containing more amount in sample
When organophosphor or carbamate chemicals for agriculture, capture mark A is just less to be combined with cholinesterase, in detection line（T lines）It is upper poly-
The indicator of collection is relatively weak if color with regard to relatively fewer, detection signal；When indicator is diffused into control line（C lines）When, refer to
Show that the capture mark B in agent is caught by the capture factor on C lines, indicator is assembled on C lines, provides corresponding detection signal.
Conversely, when the Organophosphorus and carbamate pesticides class pesticide concentration in sample is less, even without when, then can have on T lines and C lines
There is the considerable signal that can be detected.So Organophosphorus and carbamate pesticides class agricultural chemicals is achieved that using only a kind of test strips
Wide spectrum detection.
Preferably, indicator is physical tracer.This indicator can provide visual signal, such as color change.This
Sample just can intuitively judge testing result.
Preferably, capture mark A and organophosphor or carbamate chemicals for agriculture competitiveness are combined with cholinesterase.
Preferably, capture mark A is organophosphor or carbamate chemicals for agriculture, organophosphor or carbamate chemicals for agriculture
Characteristic residue, or organophosphor or carbamate chemicals for agriculture conjugate.Organophosphor or carbamate chemicals for agriculture conjugate,
What characteristic residue coupling other biological macromolecular or organic compound including organophosphor or carbamate chemicals for agriculture were formed
Macromolecular compound, it is albumen family macromolecule, polypeptide polymer, degradable high molecular polymerization that these large biological molecules can be
Thing, its role is to make organophosphor or carbamate chemicals for agriculture be easy to be coupled with indicator or another capture mark；Additionally,
The conjugate of organophosphor or carbamate chemicals for agriculture also includes the derivative of organophosphor or carbamate chemicals for agriculture, the derivative
Thing can equally be combined with cholinesterase.
Preferably, capture mark B be albumen family macromolecule, polypeptide polymer, degradable high molecular polymer, it is corresponding
The capture factor is its antibody；Albumen family macromolecule is preferably albumin, and more preferably, albumin is human serum albumins, cow's serum
Albumin, albumin rabbit serum, oralbumin.Degradable high molecular polymer is selected from carboxymethylcellulose calcium, polyethylene pyrrole
The carrier that pyrrolidone etc. is well known to those skilled in the art.
Or capture mark B is oligonucleotide sequence, the corresponding capture factor is its complementary series；
Or capture mark B and its capture factor constitute affine system pair.
Preferably, organophosphor or carbamate chemicals for agriculture are selected from parathion, fenifrothion, parathion-methyl, Rogor, water
Amine sulphur phosphorus, malathion, chlopyrifos and its conjugate.Organophosphor or carbamate chemicals for agriculture characteristic residue are selected from double sulphur
Base.
Preferably, capture mark B and its capture factor constitute affine system to being biotin-avidin system.
Preferably, indicator is concentrated capture in detection line and control line, can directly display out signal, or by one
Signal is shown after fixed treatment.Indicator can be physical tracer, chemical indicator or fluorescence indicator, particularly, physics
When developer is collaurum, macroscopic color change can be provided.Fluorescence indicator can be various fluorescins.Fluorescence
Albumen can send visible fluorescence signal in the irradiation of ultraviolet light.
A kind of agricultural chemicals method for quick, including sample is processed, sampling afterwards is added dropwise and detects examination in broad-spectrum pesticide
On paper, in the case of the instruction according to instruction, testing result is determined, wherein, broad-spectrum pesticide Test paper is as described above.
With reference to embodiment, technical scheme is further illustrated.
Embodiment 1
A kind of broad-spectrum pesticide detects gold test strip, and its indicator is collaurum, and the capture mark A of collaurum connection is to kill snout moth's larva
Sulphur phosphorus, capture mark B is BSA, and fenifrothion is coupled on BSA, and BSA is coupled on collaurum；Cholinester is loaded with T lines
Enzyme, anti-BSA antibody is loaded with C lines.
Embodiment 2
A kind of broad-spectrum pesticide detects gold test strip, and its indicator is collaurum, and the capture mark A of collaurum connection is water amine
Sulphur phosphorus-oralbumin conjugate, capture mark B is biotin, and isocarbophos-oralbumin conjugate and biotin are distinguished
It is connected on collaurum；Cholinesterase is loaded with T lines, Streptavidin is loaded with C lines.
Embodiment 3
A kind of broad-spectrum pesticide detects gold test strip, and its indicator is collaurum, and the capture mark A of collaurum connection is to poison with poison
Tick, chlopyrifos is connected on collaurum by linking group, and capture mark B is nucleotide sequence AAAAAAAAAA, is connected directly between
On collaurum, cholinesterase is loaded with T lines, nucleotide sequence TTTTTTTTTTTTTTTT is fixed with C lines.
Embodiment 4
A kind of broad-spectrum pesticide detects gold test strip, and its indicator is collaurum, and the capture mark A of collaurum connection is methyl
Parathion, capture mark B is the insulin of biotinylation, and capture mark A and capture mark B are connected on collaurum；T
Cholinesterase is loaded with line, the agarose with Avidin as part is loaded with C lines.
Embodiment 5
A kind of broad-spectrum pesticide detects gold test strip, and its indicator is red fluorescent protein, and red fluorescent protein is directly connected to
Capture mark A be Rogor, capture mark B be 2-Acetamido-2-deoxy-D-glucose, cholinesterase is loaded with T lines, be loaded with C lines
Wheat germ agglutinin.
With reference to experiment, technical scheme is further illustrated.
The agricultural chemicals aqueous solution of various concentrations is respectively configured, detects that gold test strip is examined using the broad-spectrum pesticide of embodiment 1
Survey, determine its test limit, testing result is as follows：
The test limit of table 1, different agricultural chemicals
| Pesticide name | Minimum detectability（mg/kg） | Pesticide name | Minimum detectability（mg/kg） |
| Acephatemet | 0.2 | Azodrin | 0.2 |
| Chlopyrifos | 0.2 | Aldicarb | 0.1 |
| Rogor | 0.5 | Carbofuran | 0.1 |
| Metrifonate | 0.05 | DDVP | 0.05 |
| Malathion | 0.5 | Methomyl | 0.1 |
As known from Table 1, Test paper of the invention has preferable detection limit, can meet the residues of pesticides of vegetables and fruits sample
Quick detection.
Respectively using impurity common in vegetables and fruits sample, configuration concentration is 10mg/kg standard liquids, uses embodiment 1
Broad-spectrum pesticide detection gold test strip is detected, to determine disturbed condition of the impurity to testing result.The testing result such as institute of table 2
Show：
The testing result of table 2, different impurities
| Dopant type | Concentration | Test result | Dopant type | Concentration | Test result |
| Vitamin A | 10mg/kg | It is negative | Beta carotene | 10mg/kg | It is negative |
| Vitamin B1 | 10mg/kg | It is negative | Chlorophyll | 10mg/kg | It is negative |
| Vitamin B2 | 10mg/kg | It is negative | Lutein | 10mg/kg | It is negative |
| Vitamin B3 | 10mg/kg | It is negative | Glucose | 10mg/kg | It is negative |
| Vitamin C | 10mg/kg | It is negative | Fructose | 10mg/kg | It is negative |
| Sodium chloride | 10mg/kg | It is negative |
Knowable to the experimental data of table 2, Test paper of the invention has more a lot of specificity, common in vegetables and fruits sample
Material on testing result without influence.
<110>Poly- one hundred bio tech ltd in Guangzhou
<120>A kind of broad-spectrum pesticide detects gold test strip and method
<130>
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 10
<212> DNA
<213>Artificial sequence
<400> 1
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<212> DNA
<213>Artificial sequence
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Claims (8)
1. a kind of broad-spectrum pesticide Test paper, test paper has detection line, control line and indicator, it is characterised in that：
The detection line of test paper is provided with cholinesterase, and control line is provided with the capture factor；
Capture mark A and B are connected with indicator, wherein capture mark A can be captured by cholinesterase, and capture mark B can be right
According to the capture factor capture on line；Wherein：
Capture mark A and organophosphor or carbamate chemicals for agriculture competitiveness are combined with cholinesterase, and capture mark A is organophosphor
Or the characteristic residue or organophosphor or carbamate of carbamate chemicals for agriculture, organophosphor or carbamate chemicals for agriculture
The conjugate of class agricultural chemicals；
Capture mark B is albumen family macromolecule, polypeptide polymer, carboxymethylcellulose calcium, polyvinylpyrrolidone, corresponding capture
The factor is its antibody；
Or capture mark B is oligonucleotide sequence, the corresponding capture factor is its complementary series.
2. a kind of broad-spectrum pesticide Test paper, test paper has detection line, control line and indicator, it is characterised in that：
The detection line of test paper is provided with cholinesterase, and control line is provided with the capture factor；
Capture mark A and B are connected with indicator, wherein capture mark A can be captured by cholinesterase, and capture mark B can be right
According to the capture factor capture on line；Wherein：
Capture mark A and organophosphor or carbamate chemicals for agriculture competitiveness are combined with cholinesterase, and capture mark A is organophosphor
Or the characteristic residue or organophosphor or carbamate of carbamate chemicals for agriculture, organophosphor or carbamate chemicals for agriculture
The conjugate of class agricultural chemicals；
Capture mark B and its capture factor constitute affine system pair.
3. broad-spectrum pesticide Test paper according to claim 1 and 2, it is characterised in that：Indicator is physical tracer.
4. broad-spectrum pesticide Test paper according to claim 1 and 2, it is characterised in that：Organophosphor or carbamates
Agricultural chemicals is selected from parathion, fenifrothion, parathion-methyl, Rogor, isocarbophos, malathion, chlopyrifos.
5. broad-spectrum pesticide Test paper according to claim 1 and 2, it is characterised in that：Organophosphor or carbamates
The characteristic residue of agricultural chemicals is selected from disulfide group.
6. broad-spectrum pesticide Test paper according to claim 2, it is characterised in that：Capture mark B and its capture factor structure
Into affine system to being biotin-avidin system.
7. broad-spectrum pesticide Test paper according to claim 3, it is characterised in that：Indicator is collaurum.
8. a kind of agricultural chemicals method for quick, including sample is processed, sampling afterwards is added dropwise in broad-spectrum pesticide Test paper
On, the instruction situation according to indicator determines testing result, wherein, broad-spectrum pesticide Test paper such as claim 1~7 is any
Described in one.
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| CN106324243B (en) * | 2016-08-23 | 2018-07-13 | 广东志道医药科技有限公司 | A kind of colloidal gold immuno-chromatography test paper strip and its preparation and application |
| CN106680493A (en) * | 2017-01-21 | 2017-05-17 | 上海为然环保科技有限公司 | Agricultural product pesticide residual colloidal gold detection kit |
| CN106970217B (en) * | 2017-03-22 | 2018-08-07 | 江苏美正生物科技有限公司 | A kind of immune chromatography method quantitatively detecting organophosphorus insecticide |
| CN106980019A (en) * | 2017-03-30 | 2017-07-25 | 王晓雨 | POCT quantitative detection systems |
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| CN101042405A (en) * | 2007-04-26 | 2007-09-26 | 浙江省农业科学院 | Immune colloidal gold test strip for detecting organophosphorus pesticide and method for making same |
| US8329454B2 (en) * | 2008-04-24 | 2012-12-11 | Institute Of Microchemical Technology Co., Ltd. | Device for detecting a cholinesterase-inhibiting substance comprising a hydrophilic photo-crosslinkable resin |
| CN102375055A (en) * | 2010-08-19 | 2012-03-14 | 中国人民解放军军事医学科学院微生物流行病研究所 | Multiplex detection immune chromatography chip |
| CN102539753A (en) * | 2011-12-29 | 2012-07-04 | 深圳康美生物科技股份有限公司 | Reagent kit and enzyme-linked immunochromatography for detecting various organophosphorus pesticide residues |
| CN104034886A (en) * | 2014-06-21 | 2014-09-10 | 广西博士海意信息科技有限公司 | Immune colloidal gold test strip for detecting organic phosphorus pesticide and preparation method thereof |
| CN104062440B (en) * | 2014-06-30 | 2016-05-11 | 韩雅君 | A kind of test strips of fast detecting vitamin D |
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