Mab 12H6 antibody, colloidal gold test strip and application

Abstract

The invention belongs to the technical field of virus immunodetection, and relates to detection of swine transmissible gastroenteritis virus, in particular to a Mab 12H6 antibody, a colloidal gold test strip and application. According to the invention, TGEV N protein is used as an antigen of a detection line, Mab 12H6 is used as a detection antibody combined with colloidal gold, condition optimization is carried out through different methods, the specificity and sensitivity of the methods are evaluated, and a detection method for detecting the antigen based on an N protein TGEV colloidal gold competition method is established. The established TGEV colloidal gold test strip has the advantages of strong specificity, high sensitivity, simplicity and convenience in operation, low production cost and the like, and can be applied to rapid detection of TGEV in a porcine diarrhea sample clinically.

Description

Technical Field

[0001] This invention belongs to the field of viral immunoassay technology and relates to the detection of porcine transmissible gastroenteritis virus. Background Technology

[0002] Transmissible gastroenteritis (TGEV) in pigs is a highly contagious intestinal disease caused by the transmissible gastroenteritis virus (TGEV), with vomiting and watery diarrhea as the main symptoms. Pigs of all ages are susceptible, especially piglets under 2 weeks old, which have a high mortality rate (usually 100%) after infection. However, the mortality rate is lower in piglets over 5 weeks old. In recent years, TGEV and porcine epidemic diarrhea (PED) have frequently co-occurred in Asian countries, causing significant economic losses. During TGEV outbreaks, pigs of all ages quickly become infected and exhibit symptoms of anorexia, vomiting, and diarrhea. Suckling piglets under 2-3 weeks old show more pronounced symptoms and have a high mortality rate, but the mortality rate decreases with age. Co-infection with porcine rotavirus (PRO) and PED virus is common clinically, seriously threatening the development of the pig industry. Therefore, rapid and accurate differential diagnosis of TGEV is a crucial aspect of its prevention and control.

[0003] Colloidal gold immunochromatography assay (GICA), developed in the 1990s, is widely used in animal and plant quarantine, food safety residue detection, and animal disease diagnosis due to its advantages such as simplicity, speed, low cost, and easy observation of results. Compared with traditional laboratory testing methods, colloidal gold test strips are more suitable for rapid on-site detection of large-scale samples. Application CN109232736A discloses a monoclonal antibody, pharmaceutical composition, kit, and application that specifically binds to porcine transmissible gastroenteritis virus (TGEV). This monoclonal antibody targets classic or circulating strains of TGEV, but its sensitivity needs improvement.

[0004] Therefore, a simple, rapid, and highly specific TGEV detection method is of great significance for the prevention and control of this disease in clinical practice. Summary of the Invention

[0005] This invention proposes a Mab 12H6 antibody, a colloidal gold test strip, and its application, and establishes a detection method for antigens based on the N protein TGEV colloidal gold competitive assay.

[0006] The technical solution of this invention is implemented as follows: One of the technical solutions of the present invention provides a Mab 12H6 antibody, wherein the Mab 12H6 antibody comprises a Mab 12H6 light chain and a Mab 12H6 heavy chain, the Mab 12H6 light chain comprising CDR1 as in SEQ ID No. 1, CDR2 as in SEQ ID No. 2, and CDR3 as in SEQ ID No. 3; and the Mab 12H6 heavy chain comprising CR1 as in SEQ ID No. 4, CR2 as in SEQ ID No. 5, and CR3 as in SEQ ID No. 6.

[0007] Furthermore, the aforementioned Mab 12H6-light chain also includes VL-FR1 as in SEQ ID No. 7, VL-FR2 as in SEQ ID No. 8, and VL-FR3 as in SEQ ID No. 9. The Mab 12H6-heavy chain also includes VH-FR1 as in SEQ ID No. 10, VH-FR2 as in SEQ ID No. 11, and VH-FR3 as in SEQ ID No. 12.

[0008] Specifically, the amino acid sequence of the Mab 12H6-light chain is shown in SEQ ID No. 13, and the amino acid sequence of the Mab 12H6-heavy chain is shown in SEQ ID No. 14.

[0009] The second technical solution of the present invention provides a product containing the above-mentioned Mab 12H6 antibody.

[0010] The third technical solution of this invention provides a colloidal gold test strip containing the aforementioned Mab 12H6 antibody. The colloidal gold test strip comprises a base plate and a sample pad, a conjugate pad, a detection pad, and an absorbent pad sequentially stacked on the base plate. The conjugate pad contains colloidal gold-labeled Mab 12H6 antibody. The detection pad has a control line (C line) and a detection line (T line), wherein the control line (C line) contains goat anti-mouse IgG antibody, and the detection line (T line) contains TGEN N protein. The amount of colloidal gold-labeled Mab 12H6 antibody added to the conjugate pad is 8 μL. The streak concentration of the control line (C line) is 1 mg / mL; the streak concentration of the detection line (T line) is 0.7 mg / mL.

[0011] The fourth technical solution of the present invention provides the application of the above-mentioned colloidal gold test strip in the preparation of products for detecting porcine transmissible gastroenteritis virus.

[0012] The present invention has the following beneficial effects: 1. The TGEV colloidal gold detection method established in this invention, when Mab 12H6 is selected as the gold-labeled antibody, the T-line coated N protein concentration is 0.7 mg / mL, the probe volume is 10 μL, and the detection time is 10 min, can detect viral fluid with an antigen content of 1 / 256 (5 × 10⁻⁶). 4 TCID 50 The test strip exhibits high specificity and good specificity, with a clinical sample detection accuracy rate of 96.8%. It boasts advantages such as high specificity, high sensitivity, ease of operation, and low production cost, making it suitable for rapid detection of TGEV in porcine diarrhea samples in clinical settings.

[0013] 2. The colloidal gold competitive assay strip established in this application is less affected by interfering substances in the sample and is suitable for the detection of complex clinical samples; it has better specificity and can more accurately distinguish the target substance from analogues. Attached Figure Description

[0014] To more clearly illustrate the technical solutions in the embodiments of the present invention or 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 only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0015] Figure 1 To express and identify the TGEV N protein.

[0016] Figure 2 Preparation and identification of colloidal gold solutions.

[0017] Figure 3 Determination of antibody screening for TGEV N colloidal gold probes.

[0018] Figure 4 To determine the concentration of TGEV N protein coating.

[0019] Figure 5 Determination of the amount of Mab 12H6 antibody added.

[0020] Figure 6 Determination of the amount of Mab 12H6 probe added.

[0021] Figure 7 Determining the optimal color development time for TGEV.

[0022] Figure 8 Sensitivity testing of TGEV colloidal gold test strips.

[0023] Figure 9 This is for the specific detection of TGEV colloidal gold test strips. Detailed Implementation

[0024] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0025] Unless otherwise specified, the experimental methods used in the following experimental examples are conventional methods; the materials and reagents used are commercially available unless otherwise specified.

[0026] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0027] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0028] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0029] Example 1: Expression and purification of TGEV N protein and preparation of colloidal gold solution 1. Expression and purification of TGEV N protein Take one vial of pET-32a-TGEV-N expression strain stored in the laboratory, inoculate it into 5 mL of liquid LB medium, add ampicillin (A+) to a final concentration of 50 g / mL, and incubate at 220 r / min for 8 h on a 37℃ water bath with shaker. Add the activated bacterial solution to 200 mL of LB medium containing A+ at a 1:100 ratio, and incubate on an air shaker for 6 h until the OD600nm value is between 0.6 and 0.8. Add IPTG to a final concentration of 0.8 mM, and continue incubation on a shaker for 10 h. Collect the bacterial solution, centrifuge at 8000 r / min for 10 min, discard the supernatant, and resuspend the precipitate in 20 mL of PBS. Sonicate the precipitate under ice bath conditions, sonicating for 15 min with a 2-second sonication interval of 6 seconds. Centrifuge the lysed bacterial solution at 12000 r / min for 20 min at 4℃, and collect the supernatant.

[0030] Ni-Agarose Resin packing was mixed and added to a chromatography column. The nickel column was equilibrated with TBS containing 10 mM imidazole, and then the lysed bacterial supernatant was added, allowing the liquid to drip at a rate of one drop every 3 seconds. After loading, impurities were washed away with 10 mL of TBS containing 50 mM imidazole, followed by 10 mL of TBS containing 200 mM imidazole, and the eluent was collected. The nickel column was rinsed with pure water and stored in 20% anhydrous ethanol. The eluent was placed in a dialysis bag with an 8000D pore size and dialyzed overnight in PBS at 4°C. The protein concentration was determined, and the eluent was aliquoted and stored at -80°C. The protein was identified, and the results are as follows: Figure 1 As shown, a specific band is visible around the 60 kDa molecular weight, consistent with the theoretical molecular weight of the expected recombinant protein. The target protein is expressed in both the supernatant and the precipitate. Furthermore, this protein reacts with clinically positive serum, demonstrating good immunogenicity.

[0031] 2. Preparation of colloidal gold solution Take two silanized Erlenmeyer flasks and add 50 mL of quadruple-distilled water to each. Heat them on a hot plate until boiling. Then, add 100 µL of 10% chloroauric acid concentrate and 2.5 mL of 1% trisodium citrate solution to each flask, and mix quickly. Next, rapidly add the trisodium citrate solution to the chloroauric acid solution, shake vigorously to mix, and then keep the mixture boiling on the hot plate. During boiling, the colloidal gold solution will change color from colorless to gray, black, purple, and wine red. Once the color of the colloidal gold solution remains constant, continue heating for 5 minutes to stabilize the solution.

[0032] After the colloidal gold solution cooled naturally, quadruple-distilled water was added to bring the volume to 100 mL. The colloidal gold solution was filtered through a 0.22 μm sterile filter membrane, and then the size and distribution of the colloidal gold particles were measured using a transmission electron microscope (TEM). Currently, electron microscopy is the most accurate method for determining the diameter and uniformity of colloidal gold particles. The results are as follows: Figure 2 As shown, the colloidal gold system gradually changed from colorless to gray, black, and purple during heating, eventually appearing wine-red. After the solution stabilized, heating was continued for 5 minutes to complete the colloidal stabilization treatment. The final obtained colloidal gold nanoparticle solution was a clear, homogeneous, and stable wine-red color. UV absorption spectroscopy determined the highest absorption peak of the colloidal gold solution to be around 521 nm, indicating a single absorption peak and thus a homogeneous and stable colloidal gold solution. Furthermore, using the known formula for calculating gold particle size, the prepared gold particles were estimated to be approximately 20 nm in diameter. After identification, the solution was stored at 4°C in the dark. The particle size of the colloidal gold particles in the solution was 20 nm.

[0033] Example 2: Preliminary screening of antibodies for TGEV colloidal gold test strips and selection of TGEV N protein coating concentration 1. Preliminary screening of antibodies for TGEV colloidal gold test strips Preliminary antibody screening was conducted using six existing antibodies from the laboratory: Mab 12H6, TGEV-polyclonal antibody, Mab 5B4, Mab 6H9, Mab 6H10, and Mab 13G7. These antibodies were used as probes and bound to colloidal gold. The pH was adjusted before adding an appropriate amount of antibody. The mixture was incubated with shaking for 30 min, followed by the addition of 10% BSA blocking agent and another 30 min of shaking incubation. Six probe antibodies were prepared: TGEV N protein (0.7 mg / mL) was drawn along the T line on the NC membrane; monoclonal antibody C line was drawn along the sheep anti-mouse antibody; and polyclonal antibody C line was drawn along the sheep anti-rabbit antibody. The sample was TGEV virus stock solution prepared in the laboratory (10... 7 TCID 50 The virus solution was mixed with different probe antibodies, and the preliminarily prepared test strips were inserted into the different mixtures. The color development results were recorded after ten minutes. A deeper T-line indicated better pairing, and the antibody with the deepest T-line was used for subsequent condition optimization. Results are as follows: Figure 3 As shown in the results, Mab12H6 exhibits the best performance in antigen detection, with its colorimetric signal intensity significantly higher than that of other antibodies.

[0034] The sequence of the Mab 12H6 antibody was sequenced, and the results are as follows: Mab 12H6 antibody light chain: FR1 amino acid: DIVLTQSPASLAVSLGQRATISCRAS; FR1 nucleotide: ttgtgctgacccaatctccagcttctttggctgtgtctctagggcagagggccaccatatcctgcagagccagtgaa; CDR1 amino acid: ESVDRYGNSF; CDR1 nucleotide: agttgatcgttatggcaatagttttatg; FR 2 amino acids: MHWFQQKPGQPPKLLIY; FR 2 nucleotide: cactggttccagcagaaaccaggacagccacccaaactcctcatctatcg; CDR2 amino acid: RAS; CDR2 nucleotide: tgcatccaa; FR3 amino acids: NLESGIPARFSGSGSRTDFTLTINPVEADDVATYYC; FR3 nucleotides: cctagaatctgggatccctgccaggttcagtggcagtgggtctaggacagacttcaccctcaccattaatcctgtggaggctgatgatgttgcaacctattactgt; CDR3 amino acid: QQSNEDPYT; CDR3 nucleotide: cagcaaagtaatgaggatccgtacacg; The amino acid sequence of the Mab 12H6- light chain is: DIVLTQSPASLAVSLGQRATISCRASESVDRYGNSF MHWFQQKPGQPPKLLIYRASNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPYT.

[0035] Mab 12H6 antibody heavy chain: FR1 amino acids: QVQLQQSGAELMRPGASVKLSCKAS; FR1 nucleotide: tccagctgcagcagtctggagctgagctgatgaggcctggagcttcagtgaagctgtcctgcaaggcttctggc; CR1 amino acid: GFTFTSSW; CR1 nucleotide: ttcaccttcaccagctcctggatg; FR2 amino acid: MHWAKQRPGQGLEWIGE; FR2 nucleotide: cactgggcgaagcagaggcctggacaaggccttgagtggattggagaaat; CR2 amino acid: IHPNSINT; CR2 nucleotide: tcatcctaatagtattaatactaa; FR3 amino acids: NYNEKFKGKATLTVDTSSSTAYVDLSRLTSEDSAVYYC; FR3 nucleotides: Ctacaatgaaaaattcaagggcaaggccacactgactgtagacacatcctccagcacagcctacgtggatctcagtcgtctgacatctgaggactctgcggtctattactgt; CR3 amino acid: AREEVRGLDY; CR3 nucleotide: gcaagagaggaggtacgaggtttggactac; The amino acid sequence of the Mab 12H6- heavy chain is: QVQLQQSGAELMRPGASVKLSCKASGFTFTSSW MHWAKQRPGQGLEWIGEIHPNSINTNYNEKFKGKATLTVDTSSSTAYVDLSRLTSEDSAVYYCAREEVRGLDY.

[0036] 2. Selection of TGEV N protein coating concentration Mab 12H6 was used as a monoclonal antibody bound to colloidal gold. Different concentrations of TGEV-N protein were drawn along the T line, and goat anti-mouse antibody was drawn along the C line. The antibody and probe concentrations were fixed, as was the antibody concentration along the C line. The T line concentrations were set to 0.1 mg / mL, 0.3 mg / mL, 0.5 mg / mL, 0.7 mg / mL, and 1 mg / mL, respectively. TGEV virus solution (10... 7 TCID 50 The sample and probe antibody were mixed. Different pre-prepared test strips were inserted into the mixture and incubated for 10 min. The color development results were recorded; the darkest T-line indicated the optimal streak concentration. The streak concentration with the darkest T-line was used for subsequent condition optimization. Results are as follows... Figure 4As shown, the colorimetric effect is best when the concentration of TGEV N protein coated on the detection line is 0.7 mg / mL and the concentration of goat anti-mouse IgG coated on the control line is 1 mg / mL.

[0037] Example 3: Selection of Mab 12H6 and probe dosage 1. Selection of Mab 12H6 antibody dosage Mab 12H6 antibody was used as a monoclonal antibody to bind to colloidal gold, and different amounts of antibody were added during probe preparation. 4 μL, 6 μL, 8 μL, 10 μL, and 12 μL of antibody were added to 1 mL of colloidal gold solution, respectively. The amount of probe added, the concentration of antibody at the C-line, and the concentration at the T-line were kept constant. TGEV virus solution (10... 7 TCID 50 The sample and probe antibody were mixed, and the preliminarily prepared test strips were inserted into the mixture and incubated for 10 min. The color development results were recorded; the darkest T line indicated the optimal antibody dosage. The antibody dosage with the darkest T line was used for subsequent condition optimization. Results are as follows: Figure 5 As shown, it can be observed by the naked eye that the T and C lines of the test strip show the best color development when 8 μL of antibody is added to each milliliter of colloidal gold solution. Therefore, the optimal amount of antibody added to the test strip is 8 μL.

[0038] 2. Selection of Mab 12H6 probe dosage Mab 12H6 was used as a monoclonal antibody to bind to colloidal gold. Different amounts of probe antibody were added during probe preparation. 4 μL, 6 μL, 8 μL, 10 μL, and 12 μL of probe antibody were added to 200 μL of sample solution, respectively. The amount of probe added, the concentration of antibody at the C-line, and the concentration at the T-line were kept constant. TGEV virus solution (10... 7 TCID 50 Mix the sample with different probe antibodies, insert the preliminarily prepared test strips into the mixture, and incubate for 10 min. Record the color development results, as shown below. Figure 6 As shown, a gradient of probe usage was established, with 4 μL, 6 μL, 8 μL, 10 μL, and 12 μL of probe added to 200 μL of sample solution, respectively. The figures show test strips prepared with different probe-antibody amounts. The results indicate that a probe usage of 10 μL yields the best results. The darkest T-line indicates the optimal probe-antibody dosage; the probe-antibody dosage with the darkest T-line was used for subsequent condition optimization.

[0039] Example 4: Optimization of the best color development time and sensitivity detection of TGEV colloidal gold test strips 1. Optimize the best color development time Mab 12H6 was used as a monoclonal antibody to bind to colloidal gold. A fixed amount of probe antibody was added when preparing the probe. 10 μL of probe antibody was added to 200 μL of sample solution. The amount of probe added, the concentration of antibody at the C-line, and the concentration at the T-line were kept constant. TGEV virus solution (10... 7 TCID 50 The sample and immobilized probe antibody were mixed, and the preliminarily prepared test strips were inserted into the mixture. Different incubation times were set: 8 min, 10 min, 12 min, and 15 min. The color development results were recorded; the deepest T-line indicated the optimal incubation time, and the incubation time with the deepest T-line was used for subsequent condition optimization. The results are as follows: Figure 7 As shown, when the reaction proceeds for 10 minutes, the detection results reach a stable state, at which point the color intensity of both the T line and the C line is significant and stable.

[0040] 2. Sensitivity testing of TGEV colloidal gold test strips Using Mab 12H6 antibody as a monoclonal antibody to bind to colloidal gold, a fixed amount of probe antibody was added when preparing the probe. 10 μL of probe antibody was added to 200 μL of sample solution. The amount of probe added, the concentration of antibody at the C-line, and the concentration at the T-line were kept constant. TGEV virus solution (10... 7 TCID 50 Using this as a sample, set different dilution factors to create a gradient. Fix the probe-antibody mixture, and insert the preliminarily prepared test strips into the mixture. Set the same incubation time of 10 min. Record the color development results, as shown below. Figure 8 As shown, a series of dilutions of TGEV virus stock solution were performed: the virus stock solution was serially diluted 10 times at an initial concentration of 1:2 (dilution range 1:2-1:1024), with DMEM culture medium used as a negative control. The results are shown in the figure. When the virus was diluted to a maximum of 1:256, the color intensity of the T line on the test strip was lower than that of the negative control. Therefore, the limit of detection for TGEV virus on this test strip was determined to be a dilution of 1:256. This result indicates that the test strip has high detection sensitivity and can effectively identify low concentrations of virus samples. It can be seen that a virus solution with an antigen content of 1 / 256 (5×10⁻⁶) is... 4 TCID 50 The closest multiple of T-chromosome NC and positive result is used as the cutoff point, which is also the detection limit of the lowest antigen amount.

[0041] Example 5: Specificity detection and clinical sample testing of TGEV colloidal gold test strips 1. Specificity detection of TGEV colloidal gold test strips Using Mab 12H6 antibody as a monoclonal antibody to bind to colloidal gold, a fixed amount of probe antibody was added when preparing the probe. 10 μL of probe antibody was added to 200 μL of sample solution. The amount of probe added, the concentration of antibody at the C-line, and the concentration at the T-line were kept constant. TGEV virus solution (10... 7 TCID 50 Using different viral solutions as samples, TGEV, PEDV, PSV, AstV, MRV, and GETV viral solutions were taken respectively, and probe antibodies were fixed and mixed. The preliminarily prepared test strips were then inserted into the mixture. The same incubation time of 10 min was set. The colorimetric results were recorded. The results are as follows: Figure 9 As shown, the test strip of this application can specifically detect TGEV.

[0042] 2. Clinical sample testing Take a clinical stool sample using a sterile cotton swab, centrifuge it in 1 mL of sample dilution at 12000 rpm for 10 min, collect the supernatant, and test the clinical sample using the prepared test strip.

[0043] The results are shown in Table 1: Table 1. Clinical trial results of TGEV colloidal gold test strips Table 1 shows that the TGEV colloidal gold detection method established in this invention, when Mab 12H6 is selected as the gold-labeled antibody, the T-line coated N protein concentration is 0.7 mg / mL, the probe volume is 10 μL, and the detection time is 10 min, can detect viral fluid with an antigen content of 1 / 256 (5 × 10⁻⁶). 4 TCID 50 It also has good specificity, with a clinical sample detection concordance rate of 96.8% using the RT-PCR method.

[0044] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A Mab 12H6 antibody, characterized by: The Mab 12H6 antibody comprises a Mab 12H6-light chain and a Mab 12H6-heavy chain, the Mab 12H6-light chain comprises a CDR1 as shown in SEQ ID No. 1, a CDR2 as shown in SEQ ID No. 2, and a CDR3 as shown in SEQ ID No. 3; the Mab 12H6-heavy chain comprises a CR1 as shown in SEQ ID No. 4, a CR2 as shown in SEQ ID No. 5, and a CR3 as shown in SEQ ID No.

6.

2. The Mab 12H6 antibody of claim 1, characterized in that: The Mab 12H6-light chain further comprises a VL-FR1 as shown in SEQ ID No. 7, a VL-FR2 as shown in SEQ ID No. 8, and a VL-FR3 as shown in SEQ ID No.

9.

3. The Mab 12H6 antibody of claim 2, wherein: The Mab 12H6-heavy chain further comprises a VH-FR1 as shown in SEQ ID No. 10, a VH-FR2 as shown in SEQ ID No. 11, and a VH-FR3 as shown in SEQ ID No.

12.

4. The Mab 12H6 antibody of claim 3, characterized in that: The amino acid sequence of the Mab 12H6-light chain is shown in SEQ ID No. 13, and the amino acid sequence of the Mab 12H6-heavy chain is shown in SEQ ID No.

14.

5. A product comprising the Mab 12H6 antibody of any one of claims 1-4.

6. A colloidal gold test strip, characterized in that: A product comprising the Mab 12H6 antibody of any one of claims 1-4.

7. The colloidal gold test strip according to claim 6, comprising a base plate and a sample pad, a conjugate pad, a detection pad and an absorbent pad which are sequentially laminated on the base plate, characterized in that: The Mab 12H6 antibody labeled with colloidal gold is added to the binding pad; a quality control line C and a detection line T are provided on the detection pad, wherein the quality control line C comprises a goat anti-mouse IgG antibody, and the detection line T comprises a TGEN N protein.

8. The colloidal gold test strip according to claim 7, characterized in that: The Mab 12H6 antibody labeled with colloidal gold is added to the binding pad in an amount of 8 μL, and the antibody concentration is 1 mg / mL.

9. The colloidal gold test strip according to claim 7, characterized in that: The quality control line C is drawn in a concentration of 1 mg / mL, and the detection line T is drawn in a concentration of 0.7 mg / mL.

10. Use of the colloidal gold test strip of any one of claims 6-9 in the preparation of a product for detecting porcine transmissible gastroenteritis virus.

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