Enrofloxacin and florfenicol combined detection card

Abstract

The utility model discloses an enrofloxacin and florfenicol bigeminy detection card which comprises a shell and at least two test strips, at least two detection windows and a sample adding hole are arranged on the shell, and the test strips are assembled in the shell at intervals; each test strip comprises a bottom plate as well as a sample pad, a colloidal gold conjugate pad, a nitrocellulose membrane and a water absorption pad which are arranged on the bottom plate and are sequentially connected, detection lines and a quality control line which are distributed at intervals are arranged on the nitrocellulose membrane, goat anti-mouse IgG antibodies are coated in the quality control line, florfenicol antigens are coated in the detection line of one test strip, and the detection line of the other test strip is coated in the water absorption pad. A gold-labeled antibody conjugate in the colloidal gold conjugate pad is a colloidal gold labeled florfenicol antibody; the detection line of the other test strip is coated with an enrofloxacin antigen, and the gold-labeled antibody conjugate in the colloidal gold conjugate pad is an enrofloxacin antibody labeled by colloidal gold. According to the invention, enrofloxacin and florfenicol can be rapidly and reliably detected at the same time with low cost.

Description

TECHNICAL FIELD

[0001] The utility model relates to biological detection, medical instrument technical field, especially a kind of enrofloxacin and florfenicol double detection card. BACKGROUND

[0002] Enrofloxacin is the third generation quinolone antibacterial drug of synthesis, also named ethyl cyclopropyloxacin, is quinolone antibacterial drug for special livestock and aquatic animals. It can be combined with bacterial DNA gyrase subunit A, so as to inhibit the cutting and connection function of enzyme, prevent the replication of bacterial DNA, and present antibacterial effect, with broad-spectrum antibacterial activity, with very strong permeability, with very strong killing effect to gram-negative bacteria, also with good antibacterial effect to gram-positive bacteria, oral absorption is good, blood drug concentration is high and stable, can be widely distributed in tissue, its metabolite is cyclopropyloxacin, still has strong antibacterial effect, almost all pathogenic bacteria of aquatic animals have strong antibacterial activity. It is effective for serious infection caused by drug-resistant pathogenic bacteria, and there is no cross resistance with other antibacterial agents. If people long-term consume food containing enrofloxacin exceeding standard, enrofloxacin may accumulate in the body, further harm human function, also may make human body produce drug resistance, affect cartilage development, cause deformity.

[0003] Florfenicol, also known as fluoromethanesulfonamide, is a 3-fluorine derivative of methanesulfonamide, which belongs to a new type of amidoalcohol antibiotic. Florfenicol is a new type of chloramphenicol broad-spectrum antibacterial drug for veterinary use. At present, florfenicol has been widely used in veterinary clinical prevention and treatment of bacterial diseases, such as bovine respiratory infection caused by Pasteurella multocida, mastitis, swine contagious pleuropneumonia, yellow dysentery, white dysentery, chicken colibacillosis, avian cholera, etc.

[0004] In recent years, there have been many cases of enrofloxacin and florfenicol detection exceeding standard in animal-derived food. According to the unqualified product situation report in recent years, the detection frequency of enrofloxacin and florfenicol in products with large circulation of freshwater fish is particularly prominent.

[0005] At present, the detection methods of enrofloxacin mainly include instrument detection method, fluorescence spectrophotometry, etc., but these methods need expensive instrument equipment, the detection cost is high, the operation time is long, the operation technical requirement is high and cannot be used for on-site detection. The detection methods of florfenicol mainly include high performance liquid chromatography, gas chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry and enzyme-linked immunosorbent assay. These methods have high sensitivity and good reproducibility, among which the national standard method is liquid chromatography-mass spectrometry, but the sample pretreatment is relatively complex, time-consuming, requires professional personnel and specific instrument equipment. UTILITY MODEL CONTENT

[0006] Therefore, it is necessary to provide an enrofloxacin and florfenicol double detection card.

[0007] The enrofloxacin and florfenicol double detection card is provided.

[0008] The enrofloxacin and florfenicol double detection card comprises a shell and at least two test paper strips.

[0009] In some embodiments, the test paper strip has a strip-shaped structure, and the sample pad, the colloidal gold conjugate pad, the nitrocellulose membrane and the water absorption pad all extend along the length direction of the test paper strip.

[0010] In some embodiments, the length of the test paper strip is 6cm-8cm.

[0011] In some embodiments, the width of the test paper strip is 2.5mm-4mm.

[0012] In some embodiments, the detection window has a strip-shaped window, and the two ends of each detection window are flush respectively.

[0013] In some embodiments, the detection window has a strip-shaped window, and the detection line and the quality control line are located at the middle position of the detection window.

[0014] In some embodiments, the size of the sample hole gradually decreases from the side of the outer surface of the shell to the side of the inner part of the shell.

[0015] In some embodiments, the shell comprises a shell cover and a base, the shell cover is detachably connected with the base, the test strip is assembled in the base, and the shell cover is provided with a detection window and a sample adding hole.

[0016] In some embodiments, the base is provided with a clamping groove, and the test strip is clamped in the clamping groove.

[0017] In some embodiments, the base and the shell cover are respectively provided with a limiting groove and a buckle, and the base and the shell cover are detachably connected through the limiting groove and the buckle.

[0018] In some embodiments, the shell is further provided with a detection result contrast area.

[0019] In some embodiments, the shell is provided with an identification area, and the identification area is provided with a one-dimensional code and / or a two-dimensional code capable of being scanned and identified by an external scanning device.

[0020] The enrofloxacin and florfenicol double detection card of the application detects the drug residues of enrofloxacin and florfenicol in samples by using a colloidal gold immunochromatography method, and multiple items can be simultaneously detected after one-time sample pretreatment by using the double detection card. The sample pretreatment does not need to use organic solvents and complex instruments, is less affected by sample matrix, and meets the requirements of on-site detection of water products, livestock and poultry products, and egg samples. Compared with the immunofluorescence chromatography detection card for detecting the contents of florfenicol and trimethoprim and the mass spectrometer rapid analysis of veterinary drug residues in the prior art, the operation of the application is more simple, green and environmentally friendly, one-step on-site detection, rapid detection, and low cost, and therefore the application is worthy of wide application and promotion. BRIEF DESCRIPTION OF DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the application, the following will briefly introduce the drawings needed to be used in the embodiment description. Obviously, the drawings in the following description are only some embodiments of the application, and other drawings can be obtained by those skilled in the art without creative labor.

[0022] In order to more completely understand the application and its beneficial effects, the following will be described with reference to the drawings. In the following description, the same reference numerals represent the same parts.

[0023] Figure 1 The schematic diagram of the enrofloxacin and florfenicol double detection card according to an embodiment of the application;

[0024] Figure 2 The schematic diagram of the test strip of the enrofloxacin and florfenicol double detection card according to an embodiment of the application.

[0025] Reference Signs List

[0026] 10. Enrofloxacin and florfenicol double detection card; 100. Shell; 101. Detection window; 102. Sample adding hole; 103. Detection result contrast area; 200. Test strip; 210. Bottom plate; 201. Sample pad; 202. Colloidal gold conjugate pad; 203. Nitrocellulose membrane; 204. Water absorption pad; 205. Detection line; 206. Quality control line. DETAILED DESCRIPTION

[0027] In order to make the above objectives, characteristics and advantages of the present application more apparent, concrete embodiments of the present application will be described in detail below with reference to the drawings. In the following description, a large number of specific details are set forth in order to provide a comprehensive understanding of the present application. However, the present application can be implemented in many other different ways than those described herein, and those skilled in the art can make similar improvements without departing from the spirit of the present application, and therefore the present application is not limited to the specific embodiments disclosed below.

[0028] In the description of the present application, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and therefore cannot be understood as indicating or implying that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the present application.

[0029] In the present application, unless otherwise explicitly specified and limited, the terms "mounting", "connection", "connection", "fixing" and the like should be understood broadly, for example, it can be fixedly connected, or it can be detachably connected, or it can be integrated; it can be mechanically connected, or it can be electrically connected; it can be directly connected, or it can be indirectly connected through an intermediate medium; it can be the internal communication of two elements or the interaction relationship between two elements, unless otherwise explicitly limited. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0030] In the present application, unless otherwise explicitly specified and limited, the first feature is "on" or "under" the second feature, which can be direct contact between the first and second features, or indirect contact through an intermediate medium. Moreover, the first feature "above", "above" and "above" the second feature can be directly above or obliquely above the first feature, or only indicate that the horizontal height of the first feature is higher than that of the second feature. The first feature "below", "below" and "below" the second feature can be directly below or obliquely below the first feature, or only indicate that the horizontal height of the first feature is less than that of the second feature.

[0031] In the description of the present application, the meaning of several is more than one, the meaning of multiple is more than two, greater than, less than, more than, etc. Understand as not including the number, above, below, etc. Understand as including the number. If it is described to the first, second, only for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features or implicitly indicating the order of indicated technical features.

[0032] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present application belongs. The terms used in the specification of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0033] In this paper, "optionally", "optional", "optional" means optional, that is, optional from "have" or "no" two parallel schemes. If there are multiple "optional" in a technical solution, if there is no special explanation, and there is no contradiction or mutual restriction relationship, each "optional" is independent. In this application, "optionally contains", "optionally contains" and the like, means "contains or does not contain".

[0034] In the present application, when a numerical interval (i.e., a numerical range) is involved, the distribution of the optional values in the numerical interval is considered to be continuous and includes both numerical endpoints (i.e., the minimum value and the maximum value) of the numerical interval and every value between the two numerical endpoints, unless otherwise specified. When a numerical interval refers only to integers within the numerical interval, unless otherwise specified, the two endpoint integers of the numerical range and every integer between the two endpoints are equivalent to directly listing each integer. When multiple numerical ranges are provided to describe a feature or a characteristic, the numerical ranges can be combined. In other words, unless otherwise specified, the numerical ranges disclosed in the present application should be understood to include any and all sub-ranges included therein. The "numerical" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. The "numerical interval" is intended to broadly include quantitative intervals such as percentage intervals, ratio intervals, and value intervals.

[0035] The embodiments of the present application provide an enrofloxacin and florfenicol double detection card to solve the problems of complex experimental process, the need for large instruments, long time consumption, and the need to use a large amount of organic reagents in the prior art. The enrofloxacin and florfenicol double detection card will be described below with reference to the accompanying drawings.

[0036] The enrofloxacin and florfenicol double detection card 10 provided by the embodiments of the present application is exemplarily shown in Figure 1 Figure 1 The enrofloxacin and florfenicol double detection card 10 provided by the embodiments of the present application is exemplarily shown in

[0037] In order to more clearly illustrate the structure of the enrofloxacin and florfenicol double detection card 10, the enrofloxacin and florfenicol double detection card 10 will be introduced below with reference to the accompanying drawings.

[0038] The enrofloxacin and florfenicol double detection card 10 provided by the embodiments of the present application is exemplarily shown in Figure 1 Figure 2 The enrofloxacin and florfenicol double detection card 10 provided by the embodiments of the present application is exemplarily shown in Figure 2 ​​This is a schematic diagram of a test strip 200 for an enrofloxacin and florfenicol dual test card 10 according to an embodiment of the present invention. The test strip 200 includes a base plate 210 and a sample pad 201, a colloidal gold coupling agent pad 202, a nitrocellulose membrane 203, and an absorbent pad 204, which are disposed on the base plate 210 and connected in sequence. The sample pad 201 is located at the corresponding sample application well 102. The nitrocellulose membrane 203 is provided with spaced-apart detection lines 205 and control lines 206. The detection lines 205 and control lines 206 are located at the corresponding detection windows 101. The detection lines 205 are closer to the colloidal gold coupling agent pad 202. The control lines 206 are closer to the absorbent pad 204. The control lines 206 are coated with goat anti-mouse IgG antibodies. One test strip 200 has a test line 205 coated with florfenicol antigen, and the gold-labeled antibody conjugate in the colloidal gold conjugate pad 202 is a colloidal gold-labeled florfenicol antibody. The other test strip 200 has a test line 205 coated with enrofloxacin antigen, and the gold-labeled antibody conjugate in the colloidal gold conjugate pad 202 is a colloidal gold-labeled enrofloxacin antibody.

[0039] This application presents a dual detection card 10 for enrofloxacin and florfenicol, which uses colloidal gold immunochromatography to detect drug residues of enrofloxacin and florfenicol in samples. Using this dual detection card, multiple items can be detected simultaneously after a single sample pretreatment. Pretreatment requires no organic solvents or complex instruments, is less affected by the sample matrix, and is suitable for on-demand testing of samples such as aquatic products, livestock and poultry products, and eggs. Compared to traditional methods using immunofluorescence chromatography detection cards and mass spectrometry for rapid analysis of veterinary drug residues of florfenicol and trimethoprim, this application is simpler to operate, more environmentally friendly, allows for one-step on-site testing, is rapid, and is cost-effective, thus warranting widespread application and promotion.

[0040] Preferably, see Figure 1 As shown, Figure 1 The image shows two detection windows 101 and two test strips 200. It is easy to understand that the number of detection windows 101 and test strips 200 can be set as needed.

[0041] In some of these embodiments, please refer to Figure 2 As shown, the test strip 200 has a long strip structure. The sample pad 201, colloidal gold coupling pad 202, nitrocellulose membrane 203, and absorbent pad 204 all extend along the length of the test strip 200.

[0042] In some embodiments, the length of the test strip 200 is 6cm to 8cm. The length of the test strip 200 can be set according to actual needs.

[0043] In some embodiments, the width of the test strip 200 is 2.5mm-4mm. The width of the test strip 200 can be set according to actual needs.

[0044] In some embodiments, as shown in Figure 1 Each sample well 102 is arranged at the same end of the shell 100 and aligned with the corresponding detection window 101. Preferably, as shown in Figure 1 The detection window 101 of the strip-shaped window extends along the length direction of the shell 100.

[0045] In some embodiments, as shown in Figure 1 The detection window 101 is a strip-shaped window. The detection line 205 and the quality control line 206 are located at the middle position of the detection window 101.

[0046] In some embodiments, the size of the sample well 102 gradually decreases from the side of the outer surface of the shell 100 to the side of the inner part of the shell 100. The sample well 102 with gradually decreasing size can facilitate sample addition and facilitate the smooth entry of the sample solution onto the sample pad 201.

[0047] In some embodiments, the shell 100 includes a shell cover and a base. The shell cover and the base are detachably connected. The test strip 200 is assembled in the base. The shell cover is provided with the detection window 101 and the sample well 102.

[0048] In some embodiments, the base is provided with a clamping groove. The test strip 200 is clamped in the clamping groove.

[0049] In some embodiments, the base and the shell cover are respectively provided with limiting grooves and buckles, and the base and the shell cover are detachably connected through the limiting grooves and the buckles. The buckle can be inserted into the corresponding limiting groove under the action of external force to realize fixation and clamping. A plurality of limiting grooves and buckles are arranged at both sides of the length direction of the base.

[0050] In some embodiments, as shown in Figure 1 The shell 100 is further provided with a detection result comparison area 103, and the detection result comparison area 103 includes a plurality of detection mode comparison patterns. As shown in Figure 1 The detection result comparison area 103 includes four detection mode comparison patterns. Figure 1 The detection result comparison area 103 includes four detection mode comparison patterns.

[0051] In some embodiments, an identification area is provided on the upper surface of the housing 100, and a one-dimensional code and / or a two-dimensional code capable of being scanned and identified by an external scanning device is provided in the identification area. The one-dimensional code and / or the two-dimensional code is used for scanning the record information by the scanning device. Preferably, the identification area is located at a position on the upper surface of the housing 100, for example, at one end of the upper surface of the housing 100. The identification area is not shown in the drawings.

[0052] When the above-mentioned enrofloxacin and florfenicol double detection card 10 is used for sample detection, the following steps are included:

[0053] (1) Sample pretreatment: take 1 g ± 0.1 g of sample and add it to a 15 mL centrifuge tube, accurately add 3 mL of special extraction solution to the centrifuge tube, tightly seal the bottle cap, and shake vigorously for 2 min at a frequency of 120 times / min to 150 times / min, stand for 3 min to 5 min or place in a centrifuge at room temperature at a speed of 4000 rpm to 4000 rpm for 1 min to 3 min, and the supernatant is the test solution. The extraction solution includes a phosphate solution, and the concentration of the extraction solution is 0.01 moL to 0.02 moL. The phosphate solution includes sodium phosphate (Na3PO4), potassium phosphate (K3PO4), etc.

[0054] (2) Detection:

[0055] Before testing, read the instructions carefully, and then restore the enrofloxacin and florfenicol double detection card 10 and the test solution to room temperature (20°C to 30°C).

[0056] Tear open the aluminum foil bag of the detection card, take out the detection card from the packaging bag, and place it horizontally for standby. The detection card needs to be used within 30 min to avoid moisture affecting the test results.

[0057] Use a pipette to suck up 100 μL of the above-mentioned test solution or a dropper to add 3 drops to 4 drops, and vertically drop it into the sample well 102 of the detection card. Stand for 5 min to 8 min, and according to the schematic diagram, the result is invalid for other times.

[0058] (3) Regarding the result interpretation, combined with Figure 1 shown, the main interpretation method is as follows:

[0059] Visual inspection:

[0060] Negative (-): When the detection line 205 (T line) and the quality control line 206 (C line) show purple-red bands at the same time, it indicates that the sample does not contain the test substance or its concentration is lower than the detection limit.

[0061] Positive (+): C line coloration, T line no coloration, indicating that the test substance in the sample has a residual concentration equal to or higher than the detection limit.

[0062] Invalid: C line does not appear purple red band, indicating that the incorrect operation process or the test card has been spoiled, in which case, the measured result is invalid, and the test needs to be performed again.

[0063] In summary, the enrofloxacin and florfenicol double detection card 10 of the application has the following beneficial effects:

[0064] (1) In the detection of the application, there is no organic reagent in the sample extraction solution, and no centrifugation and nitrogen blowing and other complex steps are needed, only shaking and standing are needed, and the whole sample processing process can be completed without other auxiliary equipment.

[0065] (2) The operation is simple, and the detection technician does not need much training and high skills to complete the experimental test and result interpretation well.

[0066] (3) The application uses colloidal gold immunochromatography technology, so that the product has strong specificity, high sensitivity, high accuracy, good stability and the like.

[0067] (4) The result interpretation after detection of the application is simple and intuitive, and the detection result can be obtained by comparing the detection form with the pattern, when the detection line 205 (T line) and the quality control line 206 (C line) both show purple red bands, it indicates that the sample does not contain the measured substance or its concentration is lower than the detection limit; when the detection line 205 does not show color, it indicates that the concentration of the measured substance in the sample is equal to or higher than the detection limit; when the quality control line 206 does not appear, it indicates that the operation process is incorrect or the test card has been invalid, in which case, the instruction manual should be read again, and the test should be performed again with a new test card.

[0068] (5) After pre-treatment of a sample, multiple items can be detected at the same time, the operation is efficient and fast, and it is suitable for rapid detection laboratories, entry and exit quarantine, individual breeding and production, and the like, and provides a good technical service basis for rapid screening and detection of food safety, and can meet the detection of aquatic products, livestock and poultry products and egg products.

[0069] (6) No complex reagents and expensive instruments are needed, and the on-site detection is one-step, rapid and low in cost, so it is worth wide application and promotion.

[0070] (7) The structure is simple, the sample can be detected immediately, the use is convenient, and the detection is rapid.

[0071] The technical features of the above-described embodiments can be combined arbitrarily, and in order to make the description simple, all possible combinations of the technical features in the above-described embodiments are not described, however, as long as the combination of the technical features does not exist contradictory, it should be considered as the scope of the present disclosure.

[0072] The above-described embodiments only express several implementation manners of the utility model, the description is more specific and detailed, but can not therefore be understood as the limitation of the utility model patent range. It should be pointed out that for ordinary skilled person in the art, without departing from the utility model concept, several deformations and improvements can be made, which belong to the protection range of the utility model. Therefore, the protection range of the utility model patent should be subject to the appended claims.

Claims

1. A dual test card for enrofloxacin and florfenicol, characterized in that, The application relates to a test kit, which comprises a shell and at least two test strips, the shell is provided with at least two detection windows and sample holes, the test strips are arranged in the shell at intervals, and each test strip corresponds to one detection window; the test strip comprises a bottom plate and a sample pad, a colloidal gold conjugate pad, a nitrocellulose membrane and a water absorption pad arranged on the bottom plate in sequence, the sample pad corresponds to the position of the corresponding sample hole, the nitrocellulose membrane is provided with detection lines and a quality control line arranged at intervals, the detection lines and the quality control line correspond to the positions of the corresponding detection windows, the detection lines are closer to the colloidal gold conjugate pad, the quality control line is closer to the water absorption pad, the quality control line is coated with a goat anti-mouse IgG antibody, the detection lines of one test strip are coated with florfenicol antigens, and the gold-labeled florfenicol antibody conjugate in the colloidal gold conjugate pad is a colloidal gold-labeled florfenicol antibody; the detection lines of another test strip are coated with enrofloxacin antigens, and the gold-labeled enrofloxacin antibody conjugate in the colloidal gold conjugate pad is a colloidal gold-labeled enrofloxacin antibody.

2. The enrofloxacin and florfenicol double test card according to claim 1, characterized in that, The test strip has a long strip structure, and the sample pad, the colloidal gold conjugate pad, the nitrocellulose membrane and the water absorption pad all extend along the length direction of the test strip.

3. The enrofloxacin and florfenicol double test card according to claim 1, characterized in that, The length of the test strip is 6cm-8cm. The width of the test strip is 2.5mm-4mm.

4. The enrofloxacin and florfenicol double test card according to any one of claims 1 to 3, characterized in that, The detection window is a long strip window, and the two ends of each detection window are flush with each other, and each sample hole is arranged at the same end of the shell and aligned with the corresponding detection window.

5. The Enrofloxacin and Florfenicol double test card according to any one of claims 1 to 3, characterized in that, The detection window is a long strip window, and the detection lines and the quality control line are located at the middle part of the detection window.

6. The enrofloxacin and florfenicol double test card according to any one of claims 1 to 3, characterized in that, The size of the sample hole gradually decreases from one side on the outer surface of the shell to the other side in the shell.

7. The enrofloxacin and florfenicol double test card according to any one of claims 1 to 3, characterized in that, The shell comprises a shell cover and a base, the shell cover and the base are detachably connected, the test strip is arranged in the base, and the shell cover is provided with detection windows and sample holes.

8. The enrofloxacin and florfenicol double detection card according to claim 7, characterized in that, The base is provided with a clamping groove, and the test strip is clamped in the clamping groove. The base and the shell cover are respectively provided with a limiting groove and a buckle, and the base and the shell cover are detachably connected through the limiting groove and the buckle.

9. The enrofloxacin and florfenicol double test card according to any one of claims 1 to 3, 8, characterized in that, The shell is further provided with a detection result comparison area.

10. The Enrofloxacin and Florfenicol double test card according to any one of claims 1 to 3, 8, characterized in that, The shell is provided with an identification area, and the identification area is provided with a one-dimensional code and / or a two-dimensional code which can be scanned and identified by an external scanning device.

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