CN210514332U - Detection card for detecting aflatoxin B1, zearalenone and vomitoxin - Google Patents
Detection card for detecting aflatoxin B1, zearalenone and vomitoxin Download PDFInfo
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- CN210514332U CN210514332U CN201921206938.3U CN201921206938U CN210514332U CN 210514332 U CN210514332 U CN 210514332U CN 201921206938 U CN201921206938 U CN 201921206938U CN 210514332 U CN210514332 U CN 210514332U
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Abstract
The utility model relates to a detect aflatoxin B1, zearalenone, vomit toxin's detection card, it includes the test paper strip to detect the card, the test paper strip includes the reaction membrane, the combination pad of setting in one of reaction membrane is served and the sample pad of setting on the combination pad, the length direction who follows the reaction membrane on the reaction membrane distributes in proper order and has the first detection line that is used for detecting aflatoxin B1, a second detection line for detecting zearalenone, a third detection line and quality control line for detecting vomitoxin, three kinds of mycotoxin monoclonal antibodies that the peridium has the UCP mark on the combination pad. The detection card sets up the detection line that detects aflatoxin B1, zearalenone, three kinds of mycotoxins of vomitoxin on same reaction film, simplifies the structural design and the detection card miniaturization of detection card greatly. And the detection card is marked by UCP, has high sensitivity and good stability, has no interference of organism fluorescence, and can simultaneously and quantitatively detect the residues of three mycotoxins in a sample to be detected.
Description
Technical Field
The utility model belongs to the technical field of biological short-term test, concretely relates to can be simultaneously quantitative determination fodder various raw materials and off-the-shelf aflatoxin B1, zearalenone and vomitoxin's triplex detect card.
Background
Over 300 structurally distinct mycotoxins have been discovered worldwide, of which over 20 have been isolated and identified. The annual grain loss due to mycotoxin pollution is about 5 percent. Mycotoxin contamination in feed is also a serious threat to the livestock breeding industry, not only causing economic loss, but also threatening the health of people and animals, causing mycotoxin poisoning. For example: aflatoxin B1, a carcinogen in class I regulated by the world health organization, has 68 times toxicity as that of arsenic, and is the first of the three major etiological factors of liver cancer.
The current methods for detecting mycotoxins mainly comprise: ELISA, HPLC, colloidal gold immunochromatography, Up-converting luminescence (UPT) immunochromatography, and the like. ELISA and HPLC methods have high requirements on instruments, equipment, environment, operation skills and the like, and the detection period is long and the price is high, so that the development of the feed industry in China is severely restricted. The colloidal gold immunochromatography has the problems of uneven quality, inaccurate detection result or narrow application range. The methods can only detect a single toxin at one time and cannot detect a plurality of toxins simultaneously. For example, chinese patent CN206420883U discloses a detection card for detecting aflatoxin B1, zearalenone and vomitoxin, which comprises: the kit comprises a base, wherein three detection strips and a sample loading pad are arranged on the base, each detection strip is respectively connected with the sample loading pad, each detection strip respectively comprises a detection pad, and a quality control line and a detection line which are arranged on the detection pad, the quality control line is coated with multiple antibodies of goat antibodies and mice diluted with phosphate, and the detection line is coated with antigens of aflatoxin B1, zearalenone and vomitoxin diluted with phosphate; the surface cover is arranged on the base and is provided with a sample application hole and a visual window, the sample application hole corresponds to the position of the sample loading pad, and the position of the visual window corresponds to the position of the detection pad. However, the detection card detects three kinds of moulds through three detection strips, and the structure is more complex. Moreover, aflatoxin B1, zearalenone and vomitoxin monoclonal antibodies marked by chloroauric acid are respectively sprayed on the gold pad in the patent, so that the problems of uneven detection quality, inaccurate detection result or narrow application range exist.
The up-conversion luminescence (UPT) immunochromatography is a novel mycotoxin detection technology, and has a similar principle to the colloidal gold immunochromatography: the principle of chromatographic antibody immune competition is applied. The difference lies in that UCP particles are used to replace colloidal gold particles, UCP (Up-Converting Phosphor) particles are Up-conversion luminescent materials, are optical markers, have high sensitivity and good stability, have no biological fluorescence interference, and are widely applied to the fields of grain and feed detection, disease control emergency, military equipment national safety, emergent public health events, in-vitro diagnosis and the like. However, the current up-converting luminescence (UPT) immunochromatography can only detect a single toxin at a time, and cannot detect a plurality of toxins simultaneously.
Disclosure of Invention
The utility model aims to solve the technical problem that a modified detects aflatoxin B1, zearalenone and vomitoxin's detection card is provided.
For solving the technical problem, the utility model discloses a following technical scheme:
the utility model provides a detect triple detection card of aflatoxin B1, zearalenone, vomitoxin, is in including casing and setting test paper strip in the casing, add appearance hole and observation window have been seted up respectively on the casing, the test paper strip is including being located reaction film in the casing, setting are in one of reaction film serves binding pad and setting are in sample pad on the binding pad, follow on the reaction film the length direction of reaction film distributes in proper order and has the first detection line that is used for detecting aflatoxin B1, is used for detecting the second detection line of zearalenone, is used for detecting the third detection line and the quality control line of vomitoxin, zearalenone, aflatoxin B1 monoclonal antibody that the package was had UCP mark on the binding pad.
Further, the first detection line is coated with an aflatoxin B1 artificial antigen, the second detection line is coated with a zearalenone artificial antigen, and the third detection line is coated with a vomitoxin artificial antigen.
Preferably, the artificial antigen of aflatoxin B1 is a conjugate of aflatoxin B1 and bovine serum albumin; the artificial antigen of the zearalenone is a conjugate of the zearalenone and bovine serum albumin; the vomitoxin artificial antigen is a conjugate of vomitoxin and bovine serum albumin.
Furthermore, the quality control line is coated with an anti-antibody which can be combined with an anti-aflatoxin B1 monoclonal antibody, an anti-zearalenone monoclonal antibody and an anti-vomitoxin monoclonal antibody.
Preferably, the anti-antibody is goat anti-mouse IgG.
According to some implementation aspects of the utility model, the test paper strip is still including setting up bottom plate in the casing, the reaction membrane sets up on the bottom plate, the test paper strip is still including setting up the pad that absorbs water on the other end of reaction membrane, the pad that absorbs water is close to matter accuse line, the combination pad is close to first detection line.
According to some embodiments of the utility model, in the length direction of reaction membrane, the part of combination pad, the part of the pad that absorbs water respectively with the reaction membrane overlaps, just the overlapping region length of combination pad and reaction membrane is 1~2mm, the overlapping region length of the pad that absorbs water and reaction membrane is 1~2 mm.
According to some embodiments of the invention, the bonding pad is a glass fiber membrane. The reaction membrane is a nitrocellulose membrane. The sample pad is a glass fiber membrane. The bottom plate is made of polyvinyl chloride. The absorbent pad is filter paper.
Further, the first detection line, the second detection line, the third detection line and the quality control line are parallel to each other, the distance between the adjacent first detection line, the second detection line and the third detection line is 3-4 mm, and the distance between the third detection line and the quality control line is 4.5-5.5 mm. Preferably, the distance between two adjacent first detection lines, second detection lines and third detection lines is 3.5mm, and the distance between the third detection line and the quality control line is 5 mm. The first detection line, the second detection line and the third detection line are close to each other, and the third detection line is far from the quality control line, so that pollution and interference possibly caused to the quality control line are avoided. The distance between the adjacent first detection line, the second detection line and the third detection line avoids mutual interference among the three detected toxins.
Because of the application of the technical scheme, compared with the prior art, the utility model has the following advantages:
the utility model discloses a detection card will detect aflatoxin B1, zearalenone, the three kinds of mycotoxin's of vomitoxin detection line setting on same reaction film, simplifies the structural design who detects the card greatly, and detects the card miniaturization. The detection card is marked by UCP, has high sensitivity, good stability and no biological fluorescence interference, can simultaneously detect aflatoxin B1, zearalenone and vomitoxin by only one-time sample treatment and one-time plate dropping during detection, is simple, convenient, time-saving and labor-saving, greatly shortens the sample detection time, is suitable for large-scale screening detection of a base layer, and can simultaneously quantitatively detect the residues of the mycotoxins of aflatoxin B1, zearalenone and vomitoxin in a sample to be detected.
The utility model discloses a detect aflatoxin B1, zearalenone, the minimum detection limit of vomitoxin (sensitivity) of card are 2 mug/kg, 50 mug/kg and 200 mug/kg respectively, have reached better level at the fodder detection, are applicable to all kinds of enterprises and detection mechanism. The method can be suitable for detecting various feeds, such as corn and products thereof, wheat and products thereof, broken rice, rice bran and various finished products, and has the accuracy rate of more than 90 percent compared with liquid phase detection (HPLC).
Drawings
Fig. 1 is a schematic side view of a detection card according to an embodiment of the present invention (with the housing omitted);
fig. 2 is a schematic top view of a detection card according to an embodiment of the present invention;
in the figure: 1. a sample pad; 2. a bonding pad; 3. a reaction film; 4. a base plate; 5. a water absorbent pad; 6. a first detection line; 7. a second detection line; 8. a third detection line; 9. a quality control line; 10. a sample application hole; 11. an observation window; 12. a housing.
Detailed Description
The invention is further described below with reference to the drawings in the specification:
as shown in figures 1-2, the utility model discloses a detect aflatoxin B1, zearalenone, vomitoxin's triplex detection card of an embodiment, including casing 12 and the fixed test paper strip that sets up in casing 12, set up application of sample hole 10 and observation window 11 on the casing 12 respectively. Specifically, the casing 12 is composed of an upper plastic casing and a lower plastic casing, a protruding positioning groove for fixing the test strip is arranged on the lower plastic casing, and the sample adding hole 10 and the observation window 11 are arranged on the upper plastic casing.
The test strip comprises a bottom plate 4 fixed in a shell 12 and a reaction membrane 3 arranged on the bottom plate 4, wherein a first detection line 6 for detecting aflatoxin B1, a second detection line 7 for detecting zearalenone, a third detection line 8 for detecting vomitoxin and a quality control line 9 are sequentially distributed on the reaction membrane 3 along the length direction of the reaction membrane 3, the test strip further comprises a combination pad 2 and a water absorption pad 5 which are respectively arranged at two ends of the reaction membrane 3, the combination pad 2 is arranged at one end, close to the first detection line 6, of the reaction membrane 3, the water absorption pad 5 is arranged at one end, close to the quality control line 9, of the reaction membrane 3, the test strip further comprises a sample pad 1 arranged on the combination pad 2, the combination pad 2 is positioned between the sample pad 1 and the reaction membrane 3, a sample adding hole 10 is arranged at a position corresponding to the sample pad 1, an observation window 11 is arranged at the first detection line 6, the second detection line 7, a detection window is, And the third detection line 8 and the quality control line 9 correspond to each other.
In this example, the first detection line 6 is coated with aflatoxin B1 artificial antigen, the second detection line 7 is coated with zearalenone artificial antigen, the third detection line 8 is coated with vomitoxin artificial antigen, and the quality control line 9 is coated with anti-antibody capable of combining with anti-aflatoxin B1 monoclonal antibody, anti-zearalenone monoclonal antibody and anti-vomitoxin monoclonal antibody. Specifically, the artificial antigen of aflatoxin B1 is a conjugate of aflatoxin B1 and bovine serum albumin; the artificial antigen of the zearalenone is a conjugate of the zearalenone and bovine serum albumin; the vomitoxin artificial antigen is a conjugate of vomitoxin and bovine serum albumin; the anti-antibody is goat anti-mouse IgG.
In this example, the conjugate pad 2 is a glass fiber membrane coated with UCP-labeled monoclonal antibodies to vomitoxin, zearalenone and aflatoxin B1. The reaction membrane 3 is a nitrocellulose membrane. The sample pad 1 is a glass fiber membrane. The absorbent pad 5 is filter paper.
In this example, the first detection line 6, the second detection line 7, the third detection line 8 and the quality control line 9 are parallel to each other, the distance between the adjacent first detection line 6, the second detection line 7 and the third detection line 8 is 3.5mm, and the distance between the third detection line 8 and the quality control line 9 is 5 mm. The adjacent two of the first detection line 6, the second detection line 7 and the third detection line 8 are close to each other, and the third detection line 8 is far away from the quality control line 9, so that the pollution and the interference possibly caused to the quality control line are avoided. The distance between the adjacent first detection line 6, the second detection line 7 and the third detection line 8 avoids mutual interference among the three detected toxins.
In this example, in the longitudinal direction of the reaction membrane 3, the bonding pad 2 and the water absorption pad 5 are respectively overlapped with the reaction membrane 3, and the overlapping region length of the bonding pad 2 and the reaction membrane 3 is 1-2 mm, and the overlapping region length of the water absorption pad 5 and the reaction membrane 3 is 1-2 mm.
The actual use method of the detection card comprises the following steps: and taking out the detection card from the packaging bag, sucking 100 mu L of sample solution to be detected, dripping the sample solution into the sample adding hole 10, timing for 15min after sample adding, and inserting the detection card into the up-conversion luminescence immunoassay analyzer for measurement after 15 min.
The working principle of the detection card is as follows:
vomitoxin antigen, zearalenone antigen and aflatoxin B1 antigen in a sample to be detected can be combined with UCP-labeled vomitoxin, zearalenone and aflatoxin B1 monoclonal antibodies in the chromatography process, then the chromatography is continued upwards, the antigen in the sample competes with the antigen coated by a T line (namely a detection line), the antigen of the T line can be combined with the rest of the combination, and a solid phase antigen-antibody-UCP particle compound can be formed at the position of the T line. On the C line (i.e. the quality control line), a solid phase sheep anti-mouse-antigen-antibody-UCP particle compound or a solid phase sheep anti-mouse-antibody-UCP particle compound is formed. The UCP particles emit visible light signals under exciting light, the ratio (T/C) of T line signals to C line signals is inversely proportional to the concentration of toxins in the sample, the detection card is inserted into the up-conversion luminescence immunoassay analyzer for measurement, and the content of the toxins in the sample to be detected can be read from the analyzer.
Through the above arrangement, the detection clamp has the following advantages:
(1) and (3) multi-residue quantitative detection: the detection clamp is provided with three detection lines for detecting aflatoxin B1, zearalenone and vomitoxin residues, the combination pad is marked by UCP, and the detection clamp can be matched with an up-conversion luminescence immunoassay analyzer to quantitatively detect the residues of aflatoxin B1, zearalenone and vomitoxin in a sample to be detected at the same time.
(2) The sensitivity is high: the minimum detection limits (sensitivities) of aflatoxin B1, zearalenone and vomitoxin of the detection card are respectively 2 mug/kg, 50 mug/kg and 200 mug/kg, and the detection card achieves a better level in feed detection and is suitable for various enterprises and detection mechanisms.
(3) The operation is simple and quick: the detection card has simple pretreatment and easy result judgment, can finish the detection of aflatoxin B1, zearalenone and vomitoxin residue in a feed sample only by sample treatment and plate dripping once, can finish the whole detection process within 30 minutes, and greatly shortens the sample detection time.
(4) Low cost and easy popularization: the detection card has simple production process and mature flow. Low production cost, low investment and quick effect.
(5) The accuracy is high, the practicality is strong: the detection card can be suitable for detecting various feeds, such as corn and products thereof, wheat and products thereof, broken rice, rice bran and various finished products, and has an accuracy rate of more than 90% compared with liquid phase detection (HPLC).
The above embodiments are only for illustrating the technical concept and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and to implement the present invention, so as not to limit the protection scope of the present invention, and all equivalent changes or modifications made according to the spirit of the present invention should be covered by the protection scope of the present invention.
Claims (10)
1. The utility model provides a detect aflatoxin B1, zearalenone, vomitoxin's detection card, is in including casing and setting test paper strip in the casing, add appearance hole and observation window have been seted up respectively on the casing, its characterized in that, the test paper strip is including being located reaction film in the casing, setting are in one of reaction film serves binding pad and setting and is in sample pad on the binding pad, follow on the reaction film the length direction of reaction film distributes in proper order and has the first detection line that is used for detecting aflatoxin B1, the second detection line that is used for detecting zearalenone, is used for detecting the third detection line and the quality control line of vomitoxin, the package is by aflatoxin B1, zearalenone, the vomitoxin monoclonal antibody of UCP mark on the binding pad.
2. The detection card for detecting aflatoxin B1, zearalenone, vomitoxin according to claim 1, characterized in that: the first detection line is coated with an aflatoxin B1 artificial antigen, the second detection line is coated with a zearalenone artificial antigen, and the third detection line is coated with a vomitoxin artificial antigen.
3. The detection card for detecting aflatoxin B1, zearalenone, vomitoxin according to claim 2, characterized in that: the artificial antigen of the aflatoxin B1 is a conjugate of aflatoxin B1 and bovine serum albumin; the artificial antigen of the zearalenone is a conjugate of the zearalenone and bovine serum albumin; the vomitoxin artificial antigen is a conjugate of vomitoxin and bovine serum albumin.
4. The detection card for detecting aflatoxin B1, zearalenone, vomitoxin according to claim 1, characterized in that: the quality control line is coated with an anti-antibody which can be combined with an anti-aflatoxin B1 monoclonal antibody, an anti-zearalenone monoclonal antibody and an anti-vomitoxin monoclonal antibody.
5. The detection card for detecting aflatoxin B1, zearalenone and vomitoxin according to claim 4, wherein: the anti-antibody is goat anti-mouse IgG.
6. The detection card for detecting aflatoxin B1, zearalenone and vomitoxin according to any one of claims 1 to 5, wherein: the test strip further comprises a bottom plate arranged in the shell, the reaction membrane is arranged on the bottom plate, the test strip further comprises a water absorption pad arranged at the other end of the reaction membrane, the water absorption pad is close to the quality control line, and the combination pad is close to the first detection line.
7. The detection card for detecting aflatoxin B1, zearalenone and vomitoxin according to claim 6, wherein: in the length direction of the reaction film, the part of the combination pad and the part of the water absorption pad are respectively overlapped with the reaction film, the length of an overlapping area of the combination pad and the reaction film is 1-2 mm, and the length of an overlapping area of the water absorption pad and the reaction film is 1-2 mm.
8. The detection card for detecting aflatoxin B1, zearalenone and vomitoxin according to any one of claims 1 to 5, wherein: the combination pad and the sample pad are respectively glass fiber membranes.
9. The detection card for detecting aflatoxin B1, zearalenone and vomitoxin according to any one of claims 1 to 5, wherein: the reaction membrane is a nitrocellulose membrane.
10. The detection card for detecting aflatoxin B1, zearalenone and vomitoxin according to any one of claims 1 to 5, wherein: the first detection line, the second detection line, the third detection line and the quality control line are parallel to each other, the distance between the adjacent first detection line, the second detection line and the third detection line is 3-4 mm, and the distance between the third detection line and the quality control line is 4.5-5.5 mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921206938.3U CN210514332U (en) | 2019-07-29 | 2019-07-29 | Detection card for detecting aflatoxin B1, zearalenone and vomitoxin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921206938.3U CN210514332U (en) | 2019-07-29 | 2019-07-29 | Detection card for detecting aflatoxin B1, zearalenone and vomitoxin |
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| Publication Number | Publication Date |
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| CN210514332U true CN210514332U (en) | 2020-05-12 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201921206938.3U Active CN210514332U (en) | 2019-07-29 | 2019-07-29 | Detection card for detecting aflatoxin B1, zearalenone and vomitoxin |
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| CN (1) | CN210514332U (en) |
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Effective date of registration: 20220222 Address after: 239, middle Yuewang Xingang Road, Shaxi Town, Taicang City, Suzhou City, Jiangsu Province Patentee after: ANYOU BIOTECHNOLOGY GROUP Co.,Ltd. Address before: Yuewang Taiwan funded science and Technology Innovation Industrial Park, Shaxi Town, Taicang City, Suzhou City, Jiangsu Province Patentee before: TAICANG ANYOU BIOLOGICAL TECHNOLOGY Co.,Ltd. |