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CN-122011170-A - Monoclonal antibody of porcine acute diarrhea syndrome coronavirus capsid protein and application thereof

CN122011170ACN 122011170 ACN122011170 ACN 122011170ACN-122011170-A

Abstract

The invention discloses a monoclonal antibody of a porcine acute diarrhea syndrome coronavirus capsid protein and application thereof, belonging to the technical field of biology. According to the invention, capsid protein of porcine acute diarrhea syndrome coronavirus SADS-CoV is used as a detection marker, recombinant capsid protein of SADS-CoV is expressed by a eukaryotic expression system, a monoclonal antibody against capsid protein is prepared by utilizing a hybridoma technology, and then a SADS-CoV colloidal gold rapid diagnosis test strip is prepared by utilizing a monoclonal antibody and colloidal gold test strip evaluation system. The monoclonal antibody of the porcine acute diarrhea syndrome coronavirus capsid protein provided by the invention is used for preparing an immunochromatography detection test strip, can be used for rapidly detecting the porcine acute diarrhea syndrome coronavirus, is simple and convenient, has a readily determined result, and is suitable for sites.

Inventors

  • SHI ZHENGLI
  • ZHANG WEI
  • HU BEN
  • LIU XI
  • PENG LU
  • Shu Yanni
  • CHEN YING

Assignees

  • 中国科学院武汉病毒研究所

Dates

Publication Date
20260512
Application Date
20260211

Claims (10)

  1. 1. A monoclonal antibody to a porcine acute diarrhea syndrome coronavirus capsid protein, wherein said monoclonal antibody is selected from one of the following: The monoclonal antibody PSA7 comprises a light chain and a heavy chain, wherein the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QDINNY, YTS, LQYDNLYT in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTDYV, IYPGSGST, ARGYGRGVMDY in sequence; The monoclonal antibody PSA10 comprises a light chain and a heavy chain, wherein the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are KSISKY, SGS, QQHNEYPWT in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTDYA, ISTYNGNT, AREGNYYGSNYEYAMDY in sequence; The monoclonal antibody PSA11 comprises a light chain and a heavy chain, wherein the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLVHSNGYTY, KVS, SQSTHVPLT in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSLTGYG, IWGDGST, ASNSYGNPYYGMDS in sequence.
  2. 2. The monoclonal antibody of porcine acute diarrhea syndrome coronavirus capsid protein according to claim 1, wherein the monoclonal antibody PSA7 comprises a light chain variable region having the amino acid sequence shown in SEQ ID No.1 and a heavy chain variable region having the amino acid sequence shown in SEQ ID No. 2.
  3. 3. The monoclonal antibody of porcine acute diarrhea syndrome coronavirus capsid protein according to claim 1, wherein the monoclonal antibody PSA10 comprises a light chain variable region having the amino acid sequence shown in SEQ ID No.5 and a heavy chain variable region having the amino acid sequence shown in SEQ ID No. 6.
  4. 4. The monoclonal antibody of porcine acute diarrhea syndrome coronavirus capsid protein according to claim 1, wherein the monoclonal antibody PSA11 comprises a light chain variable region having the amino acid sequence shown in SEQ ID No.9 and a heavy chain variable region having the amino acid sequence shown in SEQ ID No. 10.
  5. 5. Use of the monoclonal antibody of the porcine acute diarrhea syndrome coronavirus capsid protein of any one of claims 1-4 for preparing a porcine acute diarrhea syndrome coronavirus detection reagent.
  6. 6. An immunochromatography detection test strip for detecting porcine acute diarrhea syndrome coronavirus is characterized by comprising a sample pad, a colloidal gold pad, an NC membrane, an absorption pad and a PVC bottom plate, wherein the colloidal gold pad contains the colloidal gold-labeled porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody according to any one of claims 1-4, the NC membrane is coated with the porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody according to any one of claims 1-4, and the colloidal gold pad and the NC membrane contain different porcine acute diarrhea syndrome coronavirus N protein monoclonal antibodies.
  7. 7. The immunochromatographic test strip according to claim 6, wherein the colloidal gold pad contains a monoclonal antibody PSA11 of a capsid protein of a porcine acute diarrhea syndrome virus marked by colloidal gold, and the monoclonal antibody PSA7 of the capsid protein of the porcine acute diarrhea syndrome virus coated on the NC membrane.
  8. 8. The immunochromatographic test strip according to claim 6, wherein the colloidal gold pad contains a monoclonal antibody PSA11 of a capsid protein of a porcine acute diarrhea syndrome virus marked by colloidal gold, and the monoclonal antibody PSA10 of the capsid protein of the porcine acute diarrhea syndrome virus coated on the NC membrane.
  9. 9. The immunochromatographic test strip according to claim 6, wherein the colloidal gold pad contains a monoclonal antibody PSA10 of a capsid protein of a porcine acute diarrhea syndrome virus marked by colloidal gold, and the monoclonal antibody PSA11 of the capsid protein of the porcine acute diarrhea syndrome virus coated on the NC membrane.
  10. 10. Use of a monoclonal antibody of a porcine acute diarrhea syndrome coronavirus capsid protein according to any one of claims 1-4 or an immunochromatographic test strip according to any one of claims 6-9 for the detection or assisted detection of infection of porcine acute diarrhea syndrome coronavirus in a test sample, said use not being for the purpose of disease diagnosis.

Description

Monoclonal antibody of porcine acute diarrhea syndrome coronavirus capsid protein and application thereof Technical Field The invention belongs to the technical field of biology, and particularly relates to a porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody and a preparation method thereof, and a pathogen detection test strip based on the porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody and application thereof. Background Porcine acute diarrhea syndrome coronavirus (Swine acute diarrhoea syndrome coronavirus, SADS-CoV) was a novel coronavirus isolated in the bodies of the diarrhea piglets in guangdong in 2017. Piglets infected with SADS-CoV clinically exhibit severe diarrhea and become dehydrated and fatal within 3-5. The virus infection has resulted in death of over 27000 piglets within a short month, which brings about significant loss to the pig farming industry. Currently, SADS-CoV is identified primarily by laboratory detection means, including both antibody-based detection and viral nucleic acid detection. The ELISA method has the advantages of simple operation and lower cost, but the specificity and the sensitivity of the ELISA method need to be improved, and the RT-PCR-based detection technology provides higher sensitivity and specificity and can rapidly and accurately detect SADS-CoV. However, these techniques currently require high demands on instruments, operators, and detection environments, and the cost of reagents is also high, and are not suitable for on-site, point-of-care detection. Among them, the pathogen detection method developed for SADS-CoV is mainly nucleic acid detection, and lacks a rapid detection method which is rapid, simple and convenient, and results are easy to judge and suitable for on-site. Disclosure of Invention It is an object of the present invention to provide a monoclonal antibody to a porcine acute diarrhea syndrome coronavirus capsid protein, said monoclonal antibody being selected from one of the following: The monoclonal antibody PSA7 comprises a light chain and a heavy chain, wherein the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QDINNY, YTS, LQYDNLYT in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTDYV, IYPGSGST, ARGYGRGVMDY in sequence; The monoclonal antibody PSA10 comprises a light chain and a heavy chain, wherein the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are KSISKY, SGS, QQHNEYPWT in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTDYA, ISTYNGNT, AREGNYYGSNYEYAMDY in sequence; The monoclonal antibody PSA11 comprises a light chain and a heavy chain, wherein the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLVHSNGYTY, KVS, SQSTHVPLT in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSLTGYG, IWGDGST, ASNSYGNPYYGMDS in sequence. Further, the monoclonal antibody PSA7 comprises a light chain variable region and a heavy chain variable region, wherein the amino acid sequence of the light chain variable region is shown as SEQ ID No.1, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID No. 2; The monoclonal antibody PSA10 comprises a light chain variable region and a heavy chain variable region, wherein the amino acid sequence of the light chain variable region is shown as SEQ ID No.5, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID No. 6; the monoclonal antibody PSA11 comprises a light chain variable region and a heavy chain variable region, wherein the amino acid sequence of the light chain variable region is shown as SEQ ID No.9, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID No. 10. The preparation method of the porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody comprises the steps of immunizing a mouse with a capsid recombinant protein of the porcine acute diarrhea syndrome coronavirus, taking spleen cells, preparing hybridoma cells after fusing with SP2/0 myeloma cells, screening to obtain hybridoma cell strains which express stably and can secrete the porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody, injecting the screened hybridoma cells into a paraffin oil sensitized mouse abdominal cavity, taking mouse ascites, and purifying the ascites antibody to obtain the porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody. The second purpose of the invention is to provide the application of the monoclonal antibody of the porcine acute diarrhea syndrome coronavirus capsid protein in preparing a porcine acute diarrhea syndrome coronavirus detection reagent. The invention further provides an immunochromatography detection test strip for detecting the porcine acute diarrhea syndrome coronavirus, wherein the porcine acute diarrhea syndrome co