CN203502353U - Novel field high-sensitivity rapid Malachitegreen oxalate detection kit - Google Patents
Novel field high-sensitivity rapid Malachitegreen oxalate detection kit Download PDFInfo
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- CN203502353U CN203502353U CN201320034175.5U CN201320034175U CN203502353U CN 203502353 U CN203502353 U CN 203502353U CN 201320034175 U CN201320034175 U CN 201320034175U CN 203502353 U CN203502353 U CN 203502353U
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- malachite green
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Abstract
The utility model relates to a field high-sensitivity rapid malachite green detection kit. The detection kit is characterized by comprising a test strip and a droplet bottle, wherein a reaction tank is arranged on the test strip, and is coated with a malachite green antibody; traced fluid, positive control fluid, negative control fluid and a color reagent are contained inside the droplet bottle; the traced fluid contains pre-marked malachite green which can be specifically combined with the malachite green antibody. The rapid detection kit has the advantages of being simple in structure, easy and fast to operate, accurate in detection, high in sensitivity and the like, and capable of field detection. By adopting the rapid detection kit, simultaneous determination of a sample, positive control and negative control in an experiment can be achieved. Thus, the detection result and the validity can be accurately defined.
Description
Technical field
The utility model belongs to technical field of immunoassay, particularly relates to high sensitivity malachite green quick testing reagent box for a kind of Novel site.
Background technology
Malachite green, malachite green is a kind of green crystal with metallic luster, has another name called that Viride Nitens, salt matrix are green, peacock green, and it is fungicide, is again dyestuff, and soluble in water, solution is blue-green;
The English name of malachite green is: MalachiteGreen oxalate (another name: Malachite Green chloride; N, N, N ', N '-Tetramethyl-4,4 '-diaminotriphenylcarbenium chloride; Basic Green4), No. CAS: 569-64-2, molecular formula: C23H25ClN2, molecular weight: 364.92.Parasite, fungi or bacterium that malachite green can be used as administering fish or fish-egg infect.Be used as bacteriostatic agent or kill Amoeba agent; Concerning the aquatic/marine animals such as characin and catfish, there are the spinoffs such as height toxicity, high residue.Chemical functional group triphenylmethane in malachite green can be carcinogenic, and a lot of countries forbid, but still have fisherman to use when control infection of marine fishes fungi, also has forwarding agent to be used as sterilization, to extend the time-to-live of fish in long-distance traffic.Research finds, malachite green residence time in fish is long, and it has the spinoffs such as high toxin, high residue and carcinogenic, teratogenesis, mutagenesis, and Given this, many countries all classify malachite green as aquaculture forbidden drugs.In May, 2002, the Ministry of Agriculture listed malachite green in veterinary drug and the compound inventory > > thereof of < < food animal forbidding.
At present, the main method of detection malachite green is respectively naked eyes discriminance, liquid phase chromatography, euzymelinked immunosorbent assay (ELISA) and ELISA test strip method.Naked eyes detect, not accurate enough, lack law enforcement foundation; Liquid phase chromatography detection speed is slow, and testing cost is high, and technical requirement must be professional, particularly can not Site Detection, serious impact law enforcement dynamics and the efficiency of state functionaries; The malachite green euzymelinked immunosorbent assay (ELISA) (ELISA) that utilization has highly narrow spectrum antibody design is also current the most frequently used malachite green detection method, this method is adapted at carrying out in common laboratory, trace routine is more also needs special reading to coil instrument, must be carried out by professional and technical personnel, also be not suitable for field quick detection; ELISA test strip method, is applicable to field quick detection, but because sensitivity is low and do not carry out the positive and negative control simultaneously, easily causes the shortcomings such as false positive or false negative.
Summary of the invention
The utility model provides high sensitivity malachite green quick testing reagent box for a kind of Novel site for solving the problem existing in above-mentioned known technology.
The purpose of this utility model is to provide a kind of simple to operate, accurately quick, highly sensitive, and can carry out again high sensitivity malachite green quick testing reagent box for scene positive and that negative control detects.
The utility model is on-the-spot with high sensitivity malachite green quick testing reagent box, comprises a test-strips, traced fluid, positive control solution, negative controls; Described test-strips contains a plurality of reactive tanks, and described reactive tank is coated through malachite green antibody in advance, and confining liquid sealed; Described traced fluid, positive control solution, negative controls is to be contained in drop bottle; The malachite green that described traced fluid the contains preliminary making specifically malachite green antibody in being coated on detection groove is combined.
The utility model scene by the technical scheme that high sensitivity malachite green quick testing reagent box is taked is:
A high sensitivity malachite green quick testing reagent box for Novel site, is characterized in that: detection kit comprises test-strips and drop bottle; In drop bottle, traced fluid is housed, positive control solution, negative controls, nitrite ion; Test-strips contains three or above reactive tank, can be used for detecting positive control simultaneously, negative control and one or more sample; Reactive tank is coated through malachite green antibody antibody, and confining liquid sealed; The malachite green that traced fluid contains preliminary making, malachite green antibody or the crystal violet antibody in being coated on detection groove is combined specifically.
The utility model is on-the-spot can also adopt following technical scheme with high sensitivity malachite green quick testing reagent box:
High sensitivity malachite green quick testing reagent box for described scene, is characterized in that: detection kit also contains the drop bottle that nitrite ion is housed.
High sensitivity malachite green quick testing reagent box for described scene, is characterized in that: detection kit also contains plastic suction pipe.
High sensitivity malachite green quick testing reagent box for described scene, is characterized in: the malachite green that traced fluid contains preliminary making can directly or indirectly produce collaurum, alkaline phosphatase or the hydrogen peroxidase of naked eyes perceived color.
High sensitivity malachite green quick testing reagent box for described scene, is characterized in: nitrite ion is the substrate of the malachite green marker enzyme in traced fluid.
High sensitivity malachite green quick testing reagent box for described scene, is characterized in: positive control solution is containing malachite green, described malachite green can be in traced fluid preliminary making malachite green competitively the malachite green antibody in being coated on reactive tank be combined.
High sensitivity malachite green quick testing reagent box for described scene, is characterized in: negative controls contains damping fluid, not containing malachite green.
Advantage and the good effect that the utlity model has are:
On-the-spot with high sensitivity malachite green quick testing reagent box owing to having adopted the technical solution of the utility model, compared with prior art, have simple in structure, easy to use, do not need instrument just can detect, detection sensitivity exceeds ten times with first-class advantage than immuno-chromatographic test paper strip.
The utility model has been realized once on-the-spot with in immune detection experiment, energy is working sample simultaneously, positive control and negative control, false negative that may be hiding while having avoided using immuno-chromatographic test paper strip and the result of false inefficacy, therefore, can more correctly define testing result and validity.
Accompanying drawing explanation
Fig. 1 is that the utility model is on-the-spot with high sensitivity malachite green quick testing reagent box structural representation.
In figure, 1, test-strips, 2, traced fluid, 3, positive control solution, 4, negative controls.
Embodiment
For further understanding content of the present utility model, Characteristic, hereby exemplify following examples, and coordinate accompanying drawing to be described in detail as follows:
A high sensitivity malachite green quick testing reagent box for Novel site, comprises test-strips and drop bottle; In drop bottle, traced fluid is housed, positive control solution, negative controls; Test-strips has a plurality of reactive tanks, and reactive tank is coated through malachite green antibody, and confining liquid sealed; The malachite green that traced fluid contains collaurum preliminary making.
In traced fluid the malachite green of colloid gold label specifically the malachite green antibody in being coated on reactive tank be combined.
Positive control solution containing malachite green can with the malachite green competition of colloid gold label in traced fluid, the malachite green antibody of the malachite green that slackens colloid gold label in being coated on reactive tank is combined;
Negative controls contains negative damping fluid; In negative damping fluid, do not contain malachite green.
The specific implementation process of the present embodiment:
As shown in Figure 1, high sensitivity malachite green quick testing reagent box for a kind of Novel site, comprise: be contained in 4 test-strips (1) in plastic vacuum bag and be contained in the traced fluid (2) in drop bottle, each 1 bottle of positive control solution (3) and negative controls (4).
Test-strips is shaping plastic strip, and each test-strips contains 8 reactive tanks and can be used for measuring positive control simultaneously, a negative control and 1-6 testing sample.Reactive tank is prior after the PBS damping fluid containing 1-10ng/ml malachite green antibody is coated, with 2%BSA confining liquid, seals.
Traced fluid is that the malachite green (10ng/ml) of collaurum preliminary making is dissolved in the PBS damping fluid containing 1%BSA.
Positive control solution is containing 0.5ng/ml malachite green, is dissolved in the PBS damping fluid containing 1%BSA.
Negative controls is the PBS damping fluid containing 1%BSA, not containing malachite green.
A high sensitivity malachite green quick testing reagent box for Novel site, comprises a test-strips, traced fluid, positive control solution, negative controls, nitrite ion and plastic suction pipe; Test-strips contains a plurality of reactive tanks, and described reactive tank is coated through malachite green antibody antibody in advance, and confining liquid sealed; The malachite green antibody that traced fluid contains preliminary making.
The specific implementation process of the present embodiment:
Test-strips preparation: select to there is solid shape, contain can carrying liquid a plurality of reactive tanks, under normal temperature and pressure stable chemical nature not with the forming board of sample to be measured and the material generation chemical reaction in detection system.Described reactive tank and protide molecule have high-affinity.With 100mM Tris-HCl (pH8) damping fluid by malachite green antibody dilution to 0.1-30ug/ml, get 100 microlitres (each groove) and make an addition in the groove of test-strips, standing 1 hour or more than; After washing trough with 150 microlitres (each groove), 2% bovine serum albumin-PBS damping fluid sealing 1 hour or more than, again, after washing trough, in ℃ oven dry of 45 degree, use aluminium foil bag vacuum packaging, be placed in refrigerator standby.Described malachite green antibody can be that monoclonal malachite green antibody can be also polyclonal malachite green antibody, and described specific antibody can be buied from market also and can obtain by known preparation method for antibody.
Traced fluid preparation: by the malachite green of hydrogen peroxide enzyme labeling, release to 0.01-5000ng/ml with 1% bovine serum albumin-PBS damping fluid, be contained in after aseptic filtration in drop bottle, be placed in refrigerator standby.
Positive control solution preparation: described positive control solution is containing malachite green.1% bovine serum albumin for malachite green-PBS damping fluid is interpreted into 0.01-5ng/ml, after aseptic filtration, is contained in drop bottle, be placed in refrigerator standby.
Negative controls preparation: described negative controls contains 1% bovine serum albumin-PBS damping fluid, this feminine gender damping fluid, containing malachite green, is not contained in after aseptic filtration in drop bottle, is placed in refrigerator standby.
Nitrite ion preparation: TMB is dissolved in 50mM citrate buffer solution (pH4.5), adjusts concentration to 0.2mg/ml, then add hydrogen peroxide to 0.03%, be contained in after aseptic filtration in lucifuge drop bottle, be placed in refrigerator standby.
The testing process of high sensitivity malachite green quick testing reagent box for the present embodiment Novel site, comprises following several step:
1) application of sample reaction: get a test-strips, give each reactive tank marking, as: reactive tank 1-8; In each reactive tank, add 3 traced fluids, in reactive tank 1 and reactive tank 2, add respectively again 2 positive control solutions and 2 negative controls, with different plastic suction pipes, draw respectively different testing samples, in reactive tank 3-8, drip respectively 2 testing samples, light rolling mixes standing 5-30 minute under rear room temperature.
2) washing: outwell in reactive tank liquid and with filtered water (or tap water) rinse reactive tank 5 times or more than; After each flushing, reactive tank is swung to and fallen at thieving paper (as toilet paper) above to remove the globule in reactive tank.
3) colour developing: in each reactive tank, add 6 nitrite ions, gently shake change color in test-strips observing response groove, until in negative reaction groove, have color clearly to occur,
4) interpretation of result: relatively the shade degree of each reactive tank (positive control, negative control and each sample) is to determine that this test sample is positive or negative.
Due to positive control or positive Malachite Green can be in traced fluid the malachite green of preliminary making vie each other on reactive tank malachite green antibody be combined, and negative control can not; Therefore, positive does not develop the color under normal circumstances, negative colour developing.If the color of test sample is obviously shallow than negative control, illustrate that this test sample is positive; If the color of test sample is identical with negative control or more deeply illustrate that this test sample is negative; If the color of test sample is slightly more shallow than negative control, more positive than this test sample of the dark explanation of positive control, its malachite green concentration is lower than positive control; If the color of test sample is more shallow than positive control, illustrate that this test sample is strong positive; If the color of positive control is identical with negative control or this failure of an experiment is described more deeply.
The present embodiment has simple in structure, efficient and convenient, and detection sensitivity high is realized while working sample in once testing, positive control and negative control, thus can correctly define testing result and validity.
Claims (4)
1. a high sensitivity malachite green quick testing reagent box for Novel site, is characterized in that: described detection kit comprises test-strips and drop bottle; Described test-strips contains reactive tank coated through malachite green antibody and that sealed through confining liquid, and described drop bottle comprises: traced fluid drop bottle, positive control solution drop bottle and negative controls drop bottle.
2. according to claim 1 on-the-spot with high sensitivity malachite green quick testing reagent box, it is characterized in that: test-strips contains three or above reactive tank, can be used for detecting positive control simultaneously negative control and one or more sample.
3. according to claim 1 on-the-spot with high sensitivity malachite green quick testing reagent box, it is characterized in that: described detection kit also contains the drop bottle that nitrite ion is housed.
4. according to claim 1 on-the-spot with high sensitivity malachite green quick testing reagent box, it is characterized in that: described detection kit also contains plastic suction pipe.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320034175.5U CN203502353U (en) | 2013-01-23 | 2013-01-23 | Novel field high-sensitivity rapid Malachitegreen oxalate detection kit |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201320034175.5U CN203502353U (en) | 2013-01-23 | 2013-01-23 | Novel field high-sensitivity rapid Malachitegreen oxalate detection kit |
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| CN203502353U true CN203502353U (en) | 2014-03-26 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105628761A (en) * | 2015-06-06 | 2016-06-01 | 南华大学 | Malachite green voltammetric sensor based on imprinting recognition as well as preparation method and application thereof |
-
2013
- 2013-01-23 CN CN201320034175.5U patent/CN203502353U/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105628761A (en) * | 2015-06-06 | 2016-06-01 | 南华大学 | Malachite green voltammetric sensor based on imprinting recognition as well as preparation method and application thereof |
| CN105628761B (en) * | 2015-06-06 | 2019-05-03 | 南华大学 | Malachite green voltammetric sensor based on print identification and the preparation method and application thereof |
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