Amidoxal drug monoclonal antibody and application thereof

Abstract

The embodiment of the application discloses an amide alcohol drug monoclonal antibody and application thereof, and belongs to the technical field of biology.The amide alcohol drug monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain variable region are respectively shown as SEQ ID No.5-SEQ ID No.7, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain variable region are respectively shown as SEQ ID No.8-SEQ ID No.10.The application provides an amide alcohol drug monoclonal antibody with high titer and high sensitivity, and a test strip based on the antibody has high sensitivity, thereby providing strong technical support for the fields of amide alcohol drug residue detection, drug research and development and quality control.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to an amide alcohol drug monoclonal antibody and its application. Background Technology

[0002] Amide alcohols are a class of broad-spectrum antibiotics, mainly including chloramphenicol (CAP), thiamphenicol (TAP), florfenicol (FF), and their metabolites. They exert their antibacterial effects by inhibiting bacterial ribosome protein synthesis and are widely used in livestock, poultry, and aquaculture. Chloramphenicol, due to its severe toxicity, including irreversible aplastic anemia and gray baby syndrome, has been completely banned for use in animal food in many countries. While thiamphenicol and florfenicol have reduced toxicity, they still pose risks such as embryotoxicity and immunosuppression. Strict maximum residue limits (MRLs) have been set by various countries for these antibiotics, and long-term low-dose intake of their residues could threaten food safety and public health.

[0003] Currently, the detection of amide alcohol drug residues is mainly divided into two categories: instrumental detection and immunoassay. Instrumental detection, such as liquid chromatography-mass spectrometry (LC-MS / MS) and high performance liquid chromatography (HPLC), is accurate and sensitive, but the equipment is expensive, the pretreatment is complex, and the detection cycle is long, making it impossible to achieve rapid on-site screening. Immunoassay technology, due to its speed and portability, has become the main means of on-site screening, but its core relies on antibody performance. Existing polyclonal antibodies have poor specificity and large batch-to-batch variability, while monoclonal antibodies are mostly single-target ligands, making it difficult to simultaneously identify multiple amide alcohol drugs, and some antibodies have insufficient affinity and low sensitivity.

[0004] Therefore, developing a broad-spectrum, high-affinity, low-cross-reactivity amide alcohol monoclonal antibody to achieve simultaneous, ultra-trace rapid screening of multiple amide alcohol drugs is of great significance for improving the ability to rapidly screen for veterinary drug residues and ensuring food safety. Summary of the Invention

[0005] Therefore, embodiments of the present invention provide an amide alcohol drug monoclonal antibody and its application.

[0006] To achieve the above objectives, the embodiments of the present invention provide the following technical solutions: According to a first aspect of the present invention, the present invention provides an amide alcohol drug monoclonal antibody, comprising a heavy chain variable region and a light chain variable region, wherein the amino acid sequences of CDR1, CDR2, and CDR3 of the heavy chain variable region are shown in SEQ ID No. 5 to SEQ ID No. 7, respectively; and the amino acid sequences of CDR1, CDR2, and CDR3 of the light chain variable region are shown in SEQ ID No. 8 to SEQ ID No. 10, respectively.

[0007] Further, the amino acid sequence of the heavy chain variable region is shown in SEQ ID No. 1; the amino acid sequence of the light chain variable region is shown in SEQ ID No. 2.

[0008] Furthermore, the amide alcohol drug monoclonal antibody is prepared using chloramphenicol-carrier protein conjugate as an immunogen. The preparation method of the chloramphenicol-carrier protein conjugate includes: chloramphenicol succinate is activated by EDC / NHS and then subjected to an amidation reaction with a carrier protein to form chloramphenicol-carrier protein conjugate, wherein the carrier protein is bovine serum albumin or ovalbumin.

[0009] According to a second aspect of the present invention, the present invention provides the use of the monoclonal antibody against amide alcohols as described above in the preparation of products for detecting amide alcohols.

[0010] Furthermore, the product is a test strip.

[0011] Furthermore, the amide alcohol drugs are chloramphenicol, florfenicol, and thiamphenicol.

[0012] According to a third aspect of the present invention, the present invention provides a test strip for detecting amide alcohol drugs, wherein the conjugation pad of the test strip is coated with a monoclonal antibody against an amide alcohol drug as described above.

[0013] The embodiments of the present invention have the following advantages: This invention utilizes the complete antigen chloramphenicol-carrier protein to immunize mice, successfully obtaining high-titer, highly sensitive monoclonal antibodies against amide alcohol drugs. Based on this antibody, a highly efficient, sensitive, and specific immunoassay method for amide alcohol drugs was further established. This method not only improves the accuracy of detection but also demonstrates promising application prospects, providing strong technical support for the detection of amide alcohol drug residues, drug development, and quality control. Attached Figure Description

[0014] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other embodiments based on the provided drawings without creative effort.

[0015] Figure 1 This is an SDS-PAGE image of a monoclonal antibody against an amide alcohol drug provided by the present invention; Figure 2 The standard curve diagram provided for this invention; Figure 3A diagram showing the homology comparison results of the heavy chain gene sequence of the amyl alcohol drug monoclonal antibody provided by this invention; Figure 4 This is a homology comparison diagram of the heavy chain amino acid sequence of the amyl alcohol drug monoclonal antibody provided by the present invention; Figure 5 Homology comparison diagram of the light chain gene sequence of the amide alcohol drug monoclonal antibody provided by the present invention; Figure 6 This is a homology comparison diagram of the light chain amino acid sequence of the amyl alcohol drug monoclonal antibody provided by the present invention; Figure 7 The image shows the interpretation results of the test strip provided by this invention. Detailed Implementation

[0016] The following specific embodiments illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0017] Example 1: Preparation of artificial antigens for amide alcohol drugs The artificial antigen of amyl alcohol drugs is formed by coupling chloramphenicol with a carrier protein via succinimide ester. Its structural formula is as follows: Protein is bovine serum albumin (BSA) or ovalbumin (OVA).

[0018] The preparation method of artificial antigens for amide alcohol drugs includes the following steps: Weigh 20 mg of chloramphenicol succinate (CAP-HS) and dissolve it in 2 mL of N,N-dimethylformamide (DMF). Then, add 25 mg of EDC and 15 mg of NHS sequentially and stir at room temperature for 3 h. Dissolve 50 mg of BSA in 5 mL of 0.1 mol / L PBS (pH=7.4). Then, slowly add the chloramphenicol succinate solution dropwise to the carrier protein solution and stir at room temperature in the dark for 6 h. After the reaction is complete, dialysis with 0.1 mol / L PBS buffer (pH=7.4) to obtain immunogen CAP-BSA, which is then aliquoted and stored at -20 °C.

[0019] The preparation method of the coating antigen CAP-OVA is the same as that of CAP-BSA, except that the carrier protein BSA is replaced with an equal amount of OVA.

[0020] Example 2: Preparation of monoclonal antibodies against amide alcohol drugs 1. Animal immunization Four 6-week-old Balb / c mice, numbered #1 to #4, were selected. The immunogen CAP-BSA prepared in Example 1 was thoroughly emulsified with an equal volume of Freund's complete adjuvant and injected subcutaneously into the mice at a dose of 30 μg per mouse. Two booster immunizations were performed every two weeks for a total of two booster immunizations, with the adjuvant replaced by Freund's incomplete adjuvant. The immunization method and dose were the same as the first immunization. One week after the second booster immunization, tail blood was collected from the mice, and serum was separated for later use.

[0021] Using the CAP-OVA coating antigen prepared in Example 1 as the coating material, the antibody titer and inhibition rate of the target drug in mouse serum were detected by indirect competitive ELISA. PBS buffer was used as a blank control, and mouse serum before immunization was used as a negative control. The detection results are shown in Table 1.

[0022] Table 1

[0023] The results showed that #2 mice exhibited high and stable inhibition rates against chloramphenicol, thiamphenicol, and florfenicol at serum dilutions of 1:10000 to 1:80000, and the antibody titers met the experimental requirements. Therefore, #2 mice were selected for subsequent cell fusion experiments.

[0024] Mice #2 were aseptically sacrificed, spleens were isolated and spleen cells were prepared. These spleen cells were fused with SP2 / 0 myeloma cells and seeded into 96-well cell culture plates. The cells were cultured in HAT selective medium to screen for hybridoma cells. When cell colonies at the bottom of the wells reached 1 / 10 to 1 / 5 of the well area, positive hybridoma cells were screened using an indirect competitive ELISA method. Using 0.5 ppb chloramphenicol as a standard, the OD difference and inhibition rate of the cell supernatant were measured. The OD difference was calculated as: OD value of drug-free wells - OD value of drug-treated wells; the inhibition rate was calculated as: (OD value of drug-free wells - OD value of drug-treated wells) / OD value of drug-free wells × 100%.

[0025] The screening results of some positive hybridoma cell lines are shown in Table 2 below.

[0026] Table 2

[0027] The screening results showed that the 3A7 cell line had a high antibody expression level (drug-free OD value of cell supernatant = 2.4261) and the highest inhibition rate against 0.5 ppb chloramphenicol (83.06%), exhibiting the best antibody activity. The 3A7 cell line was cryopreserved in liquid nitrogen for subsequent large-scale preparation of monoclonal antibodies.

[0028] 2. Preparation and Identification of Monoclonal Antibodies 2.1 Monoclonal Antibody Preparation Cell resuscitation: The 3A7 positive hybridoma cell line frozen in liquid nitrogen was taken out, resuscitated using conventional cell resuscitation methods, and then transferred into cell culture flasks for expansion culture. When the cells are in the logarithmic growth phase, they are used to induce ascites in vivo.

[0029] Ascites was prepared using an in vivo induction method: Eight-week-old Balb / c mice were pretreated by intraperitoneal injection of 0.8 mL of sterile paraffin oil; seven days later, each mouse was intraperitoneally injected with 8 × 10⁸ 3A7 hybridoma cells in logarithmic growth phase. 5 Ten days after injection, ascites fluid was collected from mice under aseptic conditions. The collected ascites fluid was purified using the caprylic acid-saturated ammonium sulfate method to obtain high-purity monoclonal antibodies against amide alcohols, which were then stored at -20°C for later use.

[0030] 2.2 Monoclonal antibody purity identification The purified monoclonal antibody was analyzed by SDS-PAGE electrophoresis, such as... Figure 1 As shown, the purified antibody lanes exhibited only two characteristic bands: the IgG heavy chain (50 kDa) and the light chain (25 kDa), with no obvious impurities. Quantification by grayscale analysis confirmed the purified antibody purity to be ≥95%.

[0031] 2.3 Monoclonal antibody sensitivity detection The detection sensitivity of the purified monoclonal antibody was analyzed using an indirect competitive ELISA method: CAP-OVA prepared in Example 1 was used as the coating antigen, and chloramphenicol, florfenicol, and thiamphenicol standards at concentration gradients (0, 0.05, 0.25, 0.5, 1 ng / mL) were added respectively. Monoclonal antibody working solution diluted to an appropriate concentration was then added for competitive binding reaction. Subsequent ELISA procedures included color development and OD value measurement. The inhibition rate at each concentration was calculated, and the results are shown in Table 3 below. A standard curve was plotted. Figure 2 ), calculate the half-maximal inhibitory concentration (IC50) 50 ).

[0032] Table 3

[0033] The results showed that the three standard curves almost overlapped, indicating that the monoclonal antibody showed a high degree of consistency in recognizing chloramphenicol, thiamphenicol, and florfenicol. The standard curve R... 2 >0.99, IC 50 The concentrations were all 0.22 ng / mL, and the limit of detection was 0.05 ng / mL, indicating that the monoclonal antibody has high detection sensitivity for the three amide alcohol drugs.

[0034] 2.4 Monoclonal antibody specificity analysis The specificity of monoclonal antibodies was analyzed using an indirect competitive ELISA method: Chloramphenicol, florfenicol, and thiamphenicol (three target amide alcohol drugs), florfenicol's metabolite florfenicolamide, and other antibiotics (such as β-lactams, tetracyclines, and quinolones) were selected as analytes. The IC50 of each analyte was determined according to the sensitivity detection method described in Section 2.3. 50 value.

[0035] Using chloramphenicol as a reference (cross-reactivity = 100%), the cross-reactivity (CR) of each analyte was calculated using the following formula: CR = (chloramphenicol IC50) 50 / Analyte IC 50 ) × 100% IC50 of each analyte 50 The results of the cross-reactivity test are shown in Table 4 below.

[0036] Table 4

[0037] The results showed that the monoclonal antibody exhibited 100% cross-reactivity against the three target amyl alcohol drugs: chloramphenicol, florfenicol, and thiamphenicol, and the IC50 value was [not specified]. 50 All three antibodies have a concentration of 0.22 ng / mL, enabling broad-spectrum and highly sensitive simultaneous detection of three target drugs. They also exhibit a certain recognition ability for florfenicol's metabolite florfenicolamine (cross-reactivity rate of 22%), covering the detection of metabolite residues of the target drugs. The cross-reactivity rate with other irrelevant antibiotics is <0.005%, indicating that the antibody has extremely high specificity and the detection results are not affected by other veterinary drugs. It is suitable for the accurate detection of amide alcohol residues in actual samples (such as livestock and poultry meat, aquatic products, feed, etc.).

[0038] Example 3: Cloning of light and heavy chain variable region genes of monoclonal antibodies against amide alcohol drugs (1) Hybridoma cell culture and total RNA extraction Hybridoma cells were cultured at 37°C and 5% CO2 using RPMI 1640 complete medium at 1×10⁻⁶ oz. 7 Total RNA was extracted from cultured cells using a total RNA extraction kit.

[0039] (2) Synthesis of the first strand of cDNA Takara reverse transcription kit synthesizes cDNA.

[0040] (3) Gene amplification Design downstream primers and upstream universal primers for the Lambda, Kappa, and Heavy chains.

[0041] Upstream universal primer F: AAGCGTGGTATCAACGCAGA; Light chain downstream primer R κ :AACATTGATGTCTTTGGGGTAGAA; Light chain λ downstream primer R λ :AATCGTACACACCAGTGTGTGGG; Heavy chain downstream primer R H :AGGGATCCAGAGTTCCAGGT.

[0042] PCR was performed using the first strand of cDNA as a template in a 50 μl reaction volume.

[0043] PCR reaction system: template 3μl, upstream primer (10μM) 2.5μl, downstream primer (10μM) 2.5μl, 2×Taq enzyme 25μl, sterile water 17μl.

[0044] The landing PCR reaction conditions were as follows: 98℃ for 30 seconds; 98℃ for 15 seconds, 64℃-58℃ for 30 seconds, decreasing by 0.5℃ each time until reaching 58℃, for 10 cycles; 72℃ for 30 seconds; 98℃ for 15 seconds, 56℃ for 30 seconds, 72℃ for 30 seconds, for 15 cycles; and the program was terminated at 72℃ for 7 minutes.

[0045] (4) Cloning and screening of PCR amplification products The PCR products were subjected to 1.5% agarose gel electrophoresis. The Kappa, Lambda, and Heavy chain fragments of the antibody were recovered using a PCR product recovery kit. The fragments were inserted into the pLB vector using a pLB zero-background rapid cloning kit and transformed into DH5α competent cells (ampicillin-resistant). Recombinant positive clones were screened and sequenced.

[0046] The amino acid sequence of the heavy chain variable region of the amyl alcohol monoclonal antibody is shown in SEQ ID No. 1, and the amino acid sequence of the light chain variable region is shown in SEQ ID No. 2; the nucleotide sequence encoded by the heavy chain variable region is shown in SEQ ID No. 3, and the nucleotide sequence encoded by the light chain variable region is shown in SEQ ID No. 4.

[0047] The sequences of SEQ ID No. 1 to SEQ ID No. 4 are as follows: QVQLQQSGPQLVWPGASVKISCKASGYSFTSYRMYWVKQRPGQGLEWIGMIDPSDGETGLSQKFKDKATLTVDKSSSTAYMQFSSPTSEDSAVYYCTRRNMDFWGQGTSVTVSS (SEQ ID No. 1); DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVAWFQQKPGQSPKPLIYSASFRYSGVPDRFTGSGSGTDFTLTITNILSEDLAAYFCLQHNTYPYTFGGGTKLEIK (SEQ ID No.2); CAGGTACAACTCCAGCAGTCTGGGCCTCAGTTGGTTTGGCCTGGGGCTTCAGTGAAGATCTCCTGCAAGGCTTCTGGTTACTCATTCACCAGCTACCGGATGTACTGGGTGAAGCAGAGGCCTGGACAAGGTCTTGAGTGGATTGGCATGATTGATCCTTCCGATGGAGAAACTGGGTTAAGTCAGAAGTTCAAGGACAAGGCCACATTGACTGTTGACAAGTCCTCCAGCACAGCCTACATGCAATTCAGCAGCCCGACATCTGAGGACTCTGCGGTGTATTACTGTACAAGAAGAAATATGGACTTCTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID No.3); GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCTGTAGGAGACAGGGTCAGCGTCACCTGTAAGGCCAGTCAGAATGTGGGTACTAATGTAGCCTGGTTTCAACAGAAACCAGGGCAATCTCCTAAACCACTGATTTACTCGGCGTCCTTCCGGTACAGTGGAGTCCCTGATCGCTTCACAGGGAGTGGATCTGGGACAGATTTCACTCTCACCATCACCAATATACTGTCTGAAGACTTGGCAGCGTATTTCTGTCTGCAACATAACACCTATCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID No.4).

[0048] (5) Variable region nucleotide and amino acid sequences and homology analysis Comparative analysis in the NCBI database showed that the monoclonal antibody heavy chain variable region gene sequence had the highest homology with the mouse immunoglobulin heavy chain variable region mRNA (Sequence ID: KU256866.1), with a homology of 322 / 354 and a homology percentage of 91%. Figure 3 As shown. The amino acid sequence of the variable region of the monoclonal antibody heavy chain has the highest homology with the amino acid sequence of the variable region of the mouse immunoglobulin heavy chain (Sequence ID: AML31256.1), with a homology of 10³ / 124, representing a homology percentage of 83%. Figure 4 As shown.

[0049] The gene sequence of the light chain variable region of this monoclonal antibody showed the highest homology with the mRNA of the light chain variable region of mouse immunoglobulin κ (Sequence ID: X55042.1), with a homology of 298 / 310 and a homology percentage of 96%. Figure 5 As shown. The amino acid sequence of the variable region of the monoclonal antibody light chain has the highest homology with the mouse immunoglobulin κ light chain (Sequence ID: AAL77614.1), with a homology of 100 / 106 and a homology percentage of 94%. Figure 6 As shown.

[0050] Homology analysis of the gene and amino acid sequences encoding the light and heavy chain variable regions of monoclonal antibodies against amyl alcohol drugs showed that no sequences identical to those of the present invention were found.

[0051] The CDR regions of the light chain variable region and heavy chain variable region sequences were analyzed at https: / / www.novopro.cn / tools / cdr.html.

[0052] The amino acid sequences of CDR1, CDR2, and CDR3 in the heavy chain variable region of the amide alcohol drug are shown in SEQ ID No. 5 to SEQ ID No. 7, respectively; the amino acid sequences of CDR1, CDR2, and CDR3 in the light chain variable region are shown in SEQ ID No. 8 to SEQ ID No. 10, respectively.

[0053] The sequences of SEQ ID No. 5 to SEQ ID No. 10 are as follows: GYSFTSYR (SEQ ID No. 5); IDPSDGET (SEQ ID No. 6); TRRNMDF (SEQ ID No. 7); QNVGTN (SEQ ID No. 8); SAS (SEQ ID No. 9); LQHNTYPYT (SEQ ID No. 10).

[0054] Example 4: Preparation and application of colloidal gold immunochromatographic test strips for amide alcohol drugs 1. Preparation of colloidal gold immunochromatographic test strips for amide alcohol drugs (1) Preparation of gold-labeled pads: Colloidal gold particles with a particle size of 20 nm were mixed with monoclonal antibodies of amide alcohol drugs at a ratio of 10 μg antibody / mL colloidal gold solution, the pH was adjusted to 8.5, the mixture was sprayed onto a polyester film to form a uniform gold label layer, and the sprayed polyester film was dried at 37°C for 2 h to obtain gold-labeled pads. (2) Processing of NC membrane: Detection lines (T) and control lines (C) are pre-marked on the NC membrane with a spacing of 5 mm. The antigen CAP-OVA prepared in Example 1 is sprayed on the T line, and goat anti-rabbit IgG is sprayed on the C line. The marked NC membrane is dried at 37°C for 2 hours and then set aside. (3) Assembly of test strips: Paste the sample pad, gold label pad, NC membrane and absorbent pad in sequence onto the PVC base plate, ensuring that there is appropriate overlap between each component to form a complete liquid flow channel. Cut, package and seal the assembled test strips.

[0055] Result determination: Both the C and T lines on the test strip show color, with the T line showing much stronger color than the C line. The result is negative (-), indicating that the sample does not contain amide alcohol drugs or the color is much lower than the detection limit. If the C line of the test strip shows color, and the T line shows the same color as the C line, the T line shows a weaker color than the C line, or the T line does not show color, the result is positive (+), indicating that the concentration of amide alcohol drugs in the sample is equal to or higher than the detection limit. If neither the C line nor the T line of the test strip shows color, it indicates improper operation or that the test strip has expired.

[0056] 2. Performance evaluation of colloidal gold immunochromatographic test strips for amide alcohol drugs (1) Sensitivity The standard solutions of amide alcohols at different concentrations were tested, and the results are shown in Table 5 below.

[0057] Table 5

[0058] The results showed that the colloidal gold immunochromatographic test strip for amide alcohols prepared in this invention has high sensitivity for the detection of amide alcohols, with a detection limit of 1 ng / mL for chloramphenicol, florfenicol, and thiamphenicol, and a detection limit of 2 ng / mL for florfenicol.

[0059] (2) Specificity The colloidal gold immunochromatographic test strip for amide alcohols prepared according to this invention was used to detect chloramphenicol, florfenicol, thiamphenicol, florfenicolamide, penicillin, tetracycline, erythromycin, and kanamycin. The results showed that the target amide alcohol drugs, such as chloramphenicol, florfenicol, thiamphenicol, and florfenicolamide, were all positive, while the remaining non-target drugs were negative, indicating that the test strip provided by this invention has good specificity.

[0060] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A monoclonal antibody against an amide alcohol drug, characterized in that, It includes a heavy chain variable region and a light chain variable region, wherein the amino acid sequences of CDR1, CDR2, and CDR3 of the heavy chain variable region are shown in SEQ ID No. 5 to SEQ ID No. 7, respectively; The amino acid sequences of CDR1, CDR2, and CDR3 in the light chain variable region are shown in SEQ ID No. 8 to SEQ ID No. 10, respectively.

2. The amyl alcohol monoclonal antibody according to claim 1, characterized in that, The amino acid sequence of the heavy chain variable region is shown in SEQ ID No. 1; the amino acid sequence of the light chain variable region is shown in SEQ ID No.

2.

3. The amyl alcohol-based monoclonal antibody according to claim 1, characterized in that, The monoclonal antibody against amide alcohols is prepared using chloramphenicol-carrier protein conjugate as an immunogen. The preparation method of the chloramphenicol-carrier protein conjugate includes: chloramphenicol succinate is activated by EDC / NHS and then subjected to amidation reaction with a carrier protein to form the chloramphenicol-carrier protein conjugate, wherein the carrier protein is bovine serum albumin or ovalbumin.

4. The use of the monoclonal antibody against an amide alcohol drug as described in claim 1 in the preparation of a product for detecting an amide alcohol drug.

5. The application according to claim 4, characterized in that, The product in question is a test strip.

6. The application according to claim 4, characterized in that, The amide alcohol drugs mentioned are chloramphenicol, florfenicol, and thiamphenicol.

7. A test strip for detecting amide alcohol drugs, characterized in that, The conjugation pad of the test strip is coated with the amyl alcohol drug monoclonal antibody as described in claim 1.

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