CN211741306U - Colloidal gold test strip and colloidal gold reagent plate for detecting malachite green - Google Patents
Colloidal gold test strip and colloidal gold reagent plate for detecting malachite green Download PDFInfo
- Publication number
- CN211741306U CN211741306U CN201922497578.3U CN201922497578U CN211741306U CN 211741306 U CN211741306 U CN 211741306U CN 201922497578 U CN201922497578 U CN 201922497578U CN 211741306 U CN211741306 U CN 211741306U
- Authority
- CN
- China
- Prior art keywords
- colloidal gold
- pad
- line
- malachite green
- test strip
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims abstract description 77
- 229940107698 malachite green Drugs 0.000 title claims abstract description 38
- 238000012360 testing method Methods 0.000 title claims abstract description 36
- FDZZZRQASAIRJF-UHFFFAOYSA-M malachite green Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](C)C)C=C1 FDZZZRQASAIRJF-UHFFFAOYSA-M 0.000 title claims abstract description 34
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 22
- 239000012528 membrane Substances 0.000 claims abstract description 30
- 238000006243 chemical reaction Methods 0.000 claims abstract description 24
- 238000001514 detection method Methods 0.000 claims abstract description 22
- 239000000872 buffer Substances 0.000 claims abstract description 16
- 238000010521 absorption reaction Methods 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000007853 buffer solution Substances 0.000 claims abstract description 10
- 239000003365 glass fiber Substances 0.000 claims abstract description 9
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 8
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 8
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 8
- 238000003908 quality control method Methods 0.000 claims abstract description 8
- 241000907663 Siproeta stelenes Species 0.000 claims abstract description 7
- 238000002791 soaking Methods 0.000 claims abstract description 6
- 241000283707 Capra Species 0.000 claims abstract description 5
- 239000002250 absorbent Substances 0.000 claims description 7
- 230000002745 absorbent Effects 0.000 claims description 7
- 102000014914 Carrier Proteins Human genes 0.000 claims description 4
- 108010078791 Carrier Proteins Proteins 0.000 claims description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 3
- 229940098773 bovine serum albumin Drugs 0.000 claims description 3
- 239000010931 gold Substances 0.000 claims description 3
- 229910052737 gold Inorganic materials 0.000 claims description 3
- 229920006267 polyester film Polymers 0.000 claims description 3
- 108010058846 Ovalbumin Proteins 0.000 claims description 2
- 239000000853 adhesive Substances 0.000 claims description 2
- 230000001070 adhesive effect Effects 0.000 claims description 2
- 108060003552 hemocyanin Proteins 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 229940092253 ovalbumin Drugs 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 12
- 238000011161 development Methods 0.000 abstract description 11
- 238000012031 short term test Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 29
- 238000000034 method Methods 0.000 description 15
- 239000012488 sample solution Substances 0.000 description 9
- 239000000243 solution Substances 0.000 description 7
- 238000002965 ELISA Methods 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 239000000020 Nitrocellulose Substances 0.000 description 4
- 239000003550 marker Substances 0.000 description 4
- 229920001220 nitrocellulos Polymers 0.000 description 4
- 241000251468 Actinopterygii Species 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 238000003018 immunoassay Methods 0.000 description 3
- 238000003317 immunochromatography Methods 0.000 description 3
- 239000002207 metabolite Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- JLTDJTHDQAWBAV-UHFFFAOYSA-N N,N-dimethylaniline Chemical compound CN(C)C1=CC=CC=C1 JLTDJTHDQAWBAV-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- YADSGOSSYOOKMP-UHFFFAOYSA-N dioxolead Chemical compound O=[Pb]=O YADSGOSSYOOKMP-UHFFFAOYSA-N 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000013547 stew Nutrition 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- WZKXBGJNNCGHIC-UHFFFAOYSA-N Leucomalachite green Chemical compound C1=CC(N(C)C)=CC=C1C(C=1C=CC(=CC=1)N(C)C)C1=CC=CC=C1 WZKXBGJNNCGHIC-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000036983 biotransformation Effects 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003640 drug residue Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012764 semi-quantitative analysis Methods 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 231100000378 teratogenic Toxicity 0.000 description 1
- 230000003390 teratogenic effect Effects 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- AAAQKTZKLRYKHR-UHFFFAOYSA-N triphenylmethane Chemical compound C1=CC=CC=C1C(C=1C=CC=CC=1)C1=CC=CC=C1 AAAQKTZKLRYKHR-UHFFFAOYSA-N 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
Images
Abstract
The utility model relates to a biological detection field, in particular to detect malachite green's colloidal gold test paper strip and reagent board. The test strip comprises a bottom plate and a sample pad, a colloidal gold combination pad, a buffer pad, a reaction membrane and a water absorption pad which are sequentially connected on the bottom plate; the colloidal gold conjugate pad is coated with a conjugate of an anti-malachite green monoclonal antibody and colloidal gold; the buffer pad is prepared by soaking a glass fiber membrane in a buffer solution and then airing; a detection line T line and a quality control line C line are sequentially arranged on the reaction membrane along the chromatography direction; wherein, the T line is coated with a malachite green-carrier protein conjugate, and the C line is coated with a goat anti-mouse IgG secondary antibody. The utility model discloses a test paper strip can quick quantitative determination malachite green, and the color development is good, and accuracy and sensitivity are higher, and it is more simple and convenient to operate, is applicable to the short-term test of a large amount of samples.
Description
Technical Field
The utility model relates to a biological detection field, in particular to detect malachite green's colloidal gold test paper strip and colloidal gold reagent board.
Background
Malachite Green (MG) is a triphenylmethane dye, and is prepared by condensing benzaldehyde and N, N-dimethylaniline in hydrochloric acid or sulfuric acid to generate hidden alkali body of tetramethyldiaminotriphenylmethane, and oxidizing the hidden alkali body by lead dioxide in an acid medium. Because the chloridizing potential is similar to that of some amino acids composing enzyme, it can block the formation of protein peptide by competition action, and can be used as antibiotic and pesticide.
Malachite green has been widely used for preventing and treating saprolegniasis, branchiomycosis, ichthyophthiriasis and the like of various aquatic animals. Malachite green, however, is highly toxic, highly residual, highly carcinogenic, and highly teratogenic. After entering human or animal bodies, malachite green can be reduced and metabolized into fat-soluble recessive malachite green through biotransformation, and the toxicity is higher than that of the malachite green, thereby seriously threatening the human health. Meanwhile, the residue of malachite green also seriously influences the export of aquatic products in China, and brings a huge barrier to export trade of aquatic products in China.
The European Union and the United states have announced that the use of the fish in the culture process of economic fishes (except ornamental fishes) is forbidden. The Ministry of agriculture published No. 235 in 2002 of "highest limit of veterinary drug residue in animal food" in the publication of Ministry of agriculture in 12 months lists malachite green as a drug for prohibited use, and cannot be detected in animal food.
At present, the commonly used detection method for malachite green residue in aquatic products mainly comprises an instrument method and an immunoassay method. The instrumental methods include High Performance Liquid Chromatography (HPLC), liquid-mass chromatography (LC-MS), gas-mass chromatography, thin layer chromatography, common spectrophotometry, etc. The national Bureau of quality control in 9 months in 2005 and the State standards Commission commonly issued and implemented GB/T19857-2005 "measurement of residual amount of malachite green and crystal violet in aquatic products", which requires that the detectable rate of malachite green and its metabolites in aquatic products should not exceed 1 g/1000 tons (1 ppb). The standard specifies a liquid chromatography-tandem mass spectrometry and a high performance liquid chromatography determination method for the residual quantity of malachite green and metabolites thereof, namely recessive malachite green, crystal violet and metabolites thereof, in aquatic products, wherein the detection limit of the liquid chromatography-tandem mass spectrometry is 0.5ppb, and the detection limit of the high performance liquid chromatography is 2 ppb. The instrument method has good specificity and high accuracy, but has a certain requirement on equipment, environment, operation skill and the like, so the detection period is long, and the large-scale sample screening is not facilitated. The immunoassay method includes enzyme-linked immunosorbent assay (ELISA) and Gold Immunochromatography (GICA). The enzyme-linked immunosorbent assay has large detection amount and relatively simple operation, but the assay method also needs the assistance of laboratory equipment, has long analysis time and has certain limitation.
Colloidal Gold Immunochromatography (GICA) is a solid-phase labeled immunoassay developed in the 80 s following three major labeling techniques (fluorescein, radioisotope and enzyme). The method applies the chromatographic antibody immune competition principle, specifically detects the toxic and harmful substance residue in the sample, and has the advantages of high sensitivity, low cost, simple and quick operation and no need of special equipment for assistance.
The ELISA product for detecting the malachite green drug residue in the aquatic products provided by the markets at home and abroad has wide application and the sensitivity of 0.5-2 ppb. The method is simpler and more convenient to operate, the market cognition degree of the GICA product with lower cost is lower, the sensitivity is 2ppb, and the current detection requirement can not be met. The existing immunochromatography test strip products generally have the problems of poor color development, low sensitivity and high false positive/false negative. Therefore, the development of the colloidal gold reagent plate for detecting malachite green, which meets the requirement of detection limit and has good color development and higher sensitivity and accuracy, is of great significance.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide a detect malachite green's colloidal gold reagent board that color development is good, sensitivity and the degree of accuracy are higher.
The utility model provides a colloidal gold test strip for detecting malachite green, which comprises a bottom plate and a sample pad, a colloidal gold combination pad, a buffer pad, a reaction membrane and a water absorption pad which are connected on the bottom plate in sequence;
the colloidal gold conjugate pad is coated with a conjugate of an anti-malachite green monoclonal antibody and colloidal gold; the buffer pad is prepared by soaking a glass fiber membrane in a buffer solution and then airing; a detection line T line and a quality control line C line are sequentially arranged on the reaction membrane along the chromatography direction; wherein, the T line is coated with a malachite green-carrier protein conjugate, and the C line is coated with a goat anti-mouse IgG secondary antibody.
In the utility model discloses, buffer solution comprises PBS, Triton-100 and BSA.
Preferably, the buffer solution consists of 0.1mol/LPBS, 1% Triton-100 and 1% BSA.
The buffer pad is treated by the buffer solution, so that the pH value of the sample solution and the reaction solution of the gold-labeled antibody can be adjusted.
Preferably, the length of the cushion pad is 6 mm.
In the present invention, in the conjugate of malachite green and carrier protein, the carrier protein is bovine serum albumin, ovalbumin or hemocyanin. Preferably, the carrier protein is bovine serum albumin.
The utility model discloses in, the interval of T line and C line is 4 ~ 6 mm. Preferably, the T line and the C line are spaced apart by 5 mm.
In the colloidal gold test strip provided by the utility model, the adjacent parts of the sample pad and the colloidal gold combined pad are overlapped by 1-2 mm; the colloidal gold bonding pad is overlapped with the adjacent part of the buffer pad by 1-2 mm; the buffer pad is overlapped with the adjacent part of the reaction membrane by 1-2 mm; the reaction membrane is overlapped with the adjacent part of the water absorption pad by 1 mm-2 mm. In the colloidal gold reagent plate, the sample pad, the colloidal gold combined pad, the nitrocellulose membrane and the water absorption pad are overlapped by 1-2 mm, so that the chromatographic action is ensured to be smoothly carried out from the sample pad to the water absorption pad, and a buffer system on the sample pad can neutralize the pH value of the sample solution and ensure that effective components in the sample solution and a gold-labeled antibody on the colloidal gold combined pad are smoothly reacted in order to ensure that the sample solution and the sample pad have sufficient reaction time.
The bottom plate is made of a non-absorbent and tough material with one surface coated with non-setting adhesive, and is preferably a PVC plate; the sample pad is made of glass fibers; the colloidal gold bonding pad is made of a polyester film; the reaction membrane is a nitrocellulose membrane; the absorbent pad is filter paper.
The utility model also provides a detect malachite green's colloidal gold test paper strip and colloidal gold reagent board, including the plastic casing of above-mentioned colloidal gold test paper strip and parcel colloidal gold test paper strip, plastic casing plays fixed colloidal gold test paper strip and marks the function, is equipped with application of sample hole and observation window on the plastic casing. And dropping a sample to be detected on the sample pad of the colloidal gold test strip through the sample adding hole, wherein the observation window is used for observing the color development conditions of the C line and the T line.
Use the utility model provides a reagent board examines time measuring, wait to detect solution drop to application of sample hole S with a certain amount, wait for observing after a few minutes stew T line and C line colour depth (as shown in figure 2) in the observation window, judge according to the interpretation method that figure 3 shows and detect the sample mesomalachite green condition of remaining.
The utility model provides a test paper strip has increased the long blotter of a 6mm between colloidal gold combines to fill up and the reaction membrane, compares detection method with prior art and compares and have following advantage:
the sensitivity of the reagent strip is obviously improved, so that the detection limit is lower. The application method is simple, the color development is good, the sensitivity is high, large-scale instruments and equipment are not needed, the operation of special experiment technicians is not needed, and the method is particularly suitable for the spot check of primary supervision personnel; the prepared reagent has no toxicity or extremely low toxicity, and cannot cause harm to human bodies; the detection time is short, a large number of samples can be tested simultaneously, and the method is particularly suitable for field and field operation and easy to popularize and use; low cost, low price and convenient carrying.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a schematic view of the cross-sectional structure of a colloidal gold test strip for detecting malachite green according to the present invention;
wherein, 1 is a sample pad, 2 is a colloidal gold combined pad, 3 is a buffer pad, 4 is a reaction membrane, 5 is a detection line, 6 is a quality control line, 7 is a water absorption pad, and 8 is a bottom plate;
FIG. 2 is a top view of the colloidal gold test strip for detecting malachite green of the present invention;
FIG. 3 is a schematic diagram of the operation of the inventive gold reagent plate for detecting malachite green;
wherein, S shows a sample adding hole, 9 shows a plastic shell, and 10 shows an observation window;
fig. 4 is a schematic diagram of the test strip and the reagent plate of the present invention for judging the result, wherein T is a detection line and C is a quality control line.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
Referring to fig. 1, the cross-sectional view of the colloidal gold test strip for detecting malachite green provided by the present invention is schematically shown, and fig. 2 is a top view, and comprises a bottom plate 8 and a sample pad 1, a colloidal gold conjugate pad 2, a buffer pad 3, a reaction membrane 4 and a water absorption pad 7 which are sequentially connected to the bottom plate 8; the colloidal gold combined pad 2 is coated with a combination of an anti-malachite green monoclonal antibody and colloidal gold; the cushion pad 3 is prepared by soaking a glass fiber membrane in a buffer solution and then airing; a detection line (T line) 5 and a quality control line (C line) 6 are sequentially arranged on the reaction membrane 4 along the chromatography direction; wherein, the T line is coated with a malachite green-carrier protein conjugate, and the C line is coated with a goat anti-mouse IgG secondary antibody.
Example 1
The utility model provides a detect malachite green's colloidal gold test paper strip:
the sample pad 1 is made of glass fiber and plays a role of absorbing the pH value of the sample solution and buffering the pH value of the sample solution.
The colloidal gold conjugate pad 2 is made of a polyester film, and conjugates of an anti-malachite green monoclonal antibody and colloidal gold are labeled on the colloidal gold conjugate pad, and are sites where the active ingredients in the sample solution react with the gold-labeled antibody.
The length of the cushion pad 3 is 6mm, and the cushion pad is prepared by soaking a glass fiber membrane in a buffer solution and then airing, and the preparation method comprises the following steps: soaking the glass fiber membrane in 0.1mol/LPBS + 1% Triton-100+ 1% BSA, and air drying.
The reaction membrane 4 is a nitrocellulose membrane, and a detection line (T line) 5 and a quality control line (C line) 6 are sequentially arranged on the reaction membrane 4 along the chromatography direction; the T line is coated with a malachite green-carrier protein conjugate, the C line is coated with a goat anti-mouse IgG secondary antibody, and the main effect is to characterize the reaction result by a color which can be seen by naked eyes.
The absorbent pad 7 is a filter paper, and functions as an absorbent part to absorb the excess solution that has moved upward.
The bottom plate 8 is a PVC plate and plays a role in fixing and supporting other components of the test paper.
Wherein, the sample pad 1 and the adjacent part of the colloidal gold combined pad 2 are overlapped by 1 mm-2 mm; the adjacent parts of the colloidal gold bonding pad 2 and the buffer pad 3 are overlapped by 1-2 mm; the buffer pad 3 and the adjacent part of the reaction membrane 4 are overlapped by 1 mm-2 mm; the reaction membrane 4 is overlapped with the adjacent part of the absorbent pad 7 by 1 mm-2 mm. In the colloidal gold test strip, the sample pad 1, the colloidal gold combined pad 2, the nitrocellulose membrane 3 and the water absorption pad are overlapped by 1-2 mm, so that the chromatographic action is ensured to be smoothly carried out from the sample pad to the water absorption pad, and the buffer system on the sample pad can neutralize the pH value of the sample solution to ensure that effective components in the sample solution smoothly react with the gold-labeled antibody on the colloidal gold combined pad.
Referring to fig. 3, fig. 3 is the operation schematic diagram of the colloidal gold reagent board for detecting malachite green of the present invention, the colloidal gold reagent board includes the above colloidal gold test strip and the plastic casing 9 wrapping the colloidal gold test strip, and the plastic casing 9 is provided with the sample adding hole S and the observation window 10. The plastic shell 9 has the functions of fixing the colloidal gold test strip and marking, a sample to be detected is dripped on the sample pad 1 of the colloidal gold test strip through the sample adding hole S, and the color development conditions of the C line and the T line are observed through the observation window 10.
Use the utility model provides a reagent board examines time measuring, wait to detect solution drop to application of sample hole S with a certain amount, stew and observe after waiting for a few minutes T line and C line colour depth (as shown in fig. 3) in the observation window, judge according to the interpretation method that fig. 4 shows and detect the sample in the green circumstances of remaining of malachite, the interpretation method is: the color development of the T line is stronger or the same as that of the C line, and the T line is negative; the T line is positive without color development or weaker than the C line; the C line did not develop color, and was not effective regardless of whether T developed color or not. The identification method is based on the principle of competitive immune colloidal gold chromatography, colloidal gold is used as a tracer marker to be combined with protein molecules (antibodies) to form a gold-labeled antibody, an antigen and a substance to be detected compete to combine with the gold-labeled antibody, a color reaction is generated after the antigen and the substance to be detected are intercepted and is gathered on a detection band, a free marker crosses the detection band, the purpose of separating the free marker from the combined marker is achieved, and qualitative or semi-quantitative analysis is performed according to the purpose.
Example 2
The experiment compares the sensitivity of the buffered and unbuffered reagent plates. The malachite green standard solution was added to the negative solution to final concentrations of 0.1, 0.25, 0.5 and 1 μ g/kg, the buffered and unbuffered reagent plates were titrated separately, and the assay data and assay results were determined using a gold colloid reader, the experimental results are shown in table 1.
TABLE 1 results of sensitivity test on reagent plate
The result shows, the utility model provides a colloidal gold test paper strip has increased the blotter between colloidal gold combination pad and reaction film, has obviously improved sensitivity, and reagent board detection limit is lower, the color development is better.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (8)
1. A colloidal gold test strip for detecting malachite green comprises a base plate and a sample pad, a colloidal gold combination pad, a buffer pad, a reaction membrane and a water absorption pad which are sequentially connected on the base plate;
the colloidal gold conjugate pad is coated with a conjugate of an anti-malachite green monoclonal antibody and colloidal gold; the buffer pad is prepared by soaking a glass fiber membrane in a buffer solution and then airing; a detection line T line and a quality control line C line are sequentially arranged on the reaction membrane along the chromatography direction; wherein, the T line is coated with a malachite green-carrier protein conjugate, and the C line is coated with a goat anti-mouse IgG secondary antibody.
2. The colloidal gold test strip of claim 1, wherein the malachite green-carrier protein conjugate comprises a carrier protein selected from the group consisting of bovine serum albumin, ovalbumin, and hemocyanin.
3. The colloidal gold test strip of claim 1, wherein the length of the buffer pad is 6 mm.
4. The colloidal gold test strip of claim 1, wherein the T line and the C line are separated by 4-6 mm.
5. The colloidal gold test strip of claim 1, wherein the sample pad overlaps the gold conjugate pad by 1-2 mm; the colloidal gold bonding pad is overlapped with the adjacent part of the buffer pad by 1-2 mm; the buffer pad is overlapped with the adjacent part of the reaction membrane by 1-2 mm; the reaction membrane is overlapped with the adjacent part of the water absorption pad by 1 mm-2 mm.
6. The colloidal gold test strip of claim 1, wherein the base plate is a non-absorbent, tough material coated with a non-setting adhesive on one side; the sample pad and the reaction membrane are made of glass fibers; the colloidal gold bonding pad is made of a polyester film; the absorbent pad is filter paper.
7. A colloidal gold reagent plate for detecting malachite green, which comprises the colloidal gold test strip of any one of claims 1-6 and a plastic shell for wrapping the colloidal gold test strip.
8. A colloidal gold reagent plate according to claim 7, wherein the plastic casing is provided with a sample application hole and a viewing window.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201922497578.3U CN211741306U (en) | 2019-12-31 | 2019-12-31 | Colloidal gold test strip and colloidal gold reagent plate for detecting malachite green |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201922497578.3U CN211741306U (en) | 2019-12-31 | 2019-12-31 | Colloidal gold test strip and colloidal gold reagent plate for detecting malachite green |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN211741306U true CN211741306U (en) | 2020-10-23 |
Family
ID=72867414
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201922497578.3U Active CN211741306U (en) | 2019-12-31 | 2019-12-31 | Colloidal gold test strip and colloidal gold reagent plate for detecting malachite green |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN211741306U (en) |
-
2019
- 2019-12-31 CN CN201922497578.3U patent/CN211741306U/en active Active
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Han et al. | An octuplex lateral flow immunoassay for rapid detection of antibiotic residues, aflatoxin M1 and melamine in milk | |
| Wang et al. | Novel fabrication of immunochromatographic assay based on up conversion phosphors for sensitive detection of clenbuterol | |
| EP0677170B1 (en) | Test strip for immunoassays | |
| Li et al. | A sensitive immunochromatographic assay using colloidal gold–antibody probe for rapid detection of pharmaceutical indomethacin in water samples | |
| CN1191603A (en) | Competitive immunoassay device | |
| GB1603001A (en) | Determination of immunogens and antibodies | |
| CN109374907B (en) | Colistin colloidal gold detection kit and application thereof | |
| CN109061134B (en) | Fluorescent microsphere-colloidal gold double-color-development qualitative and quantitative immunochromatographic test strip for detecting salbutamol and preparation method thereof | |
| CN107561273A (en) | A kind of time-resolved fluorescence test strips for detecting Aflatoxins M1 and its application | |
| Xu et al. | Functional up-conversion nanoparticle-based immunochromatography assay for simultaneous and sensitive detection of residues of four tetracycline antibiotics in milk | |
| CN202794177U (en) | Kit for enzyme-linked immune-chromatography | |
| CN107589265A (en) | A kind of time-resolved fluorescence test strips for detecting aflatoxin B1 and its application | |
| CN109061205B (en) | Fluorescent microsphere-colloidal gold double-color-development qualitative and quantitative immunochromatographic test strip for detecting ractopamine and preparation method thereof | |
| CN211741306U (en) | Colloidal gold test strip and colloidal gold reagent plate for detecting malachite green | |
| CN212031502U (en) | Lateral flow reagent strip with indicating area and detection card comprising same | |
| CN110596378A (en) | Multichannel universal chromatography method for detecting small molecules, test strip and kit | |
| JPS63210664A (en) | Dry test piece for device using oxygen demand detection system and detecting method of analytic component in fluid to be inspected | |
| CN111474359B (en) | Preparation method and application of colloidal gold immunochromatographic test strip for multi-joint detection | |
| CN112763728B (en) | Detection method and test strip for detecting concentration of liquid sample | |
| CN211785569U (en) | Colloidal gold test strip and colloidal gold reagent plate for detecting chloramphenicol | |
| CN112986567A (en) | Reagent component ratio and preparation method of multi-project rapid quantitative drug detection box | |
| CN106370843A (en) | Quick measurement method of lean meat powder on the basis of gold magnetic immuno-chromatography | |
| JPH02120665A (en) | Analyzer/reader | |
| CN210514332U (en) | Detection card for detecting aflatoxin B1, zearalenone and vomitoxin | |
| CN111141907A (en) | Florfenicol rapid detection kit and detection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231121 Address after: Room 451, Building 4, Wenhua Road, Jiande Economic Development Zone, Jiande City, Hangzhou City, Zhejiang Province, 311618 Patentee after: Zhejiang Zhikang Biotechnology Co.,Ltd. Address before: Building B, 10th Floor, Union Center South Zone, No. 481 Minhe Road, Xiaoshan District, Hangzhou City, Zhejiang Province, 311215 Patentee before: HANGZHOU NANKAI BIOTECH Co.,Ltd. |
|
| TR01 | Transfer of patent right |