33 results about "African swine fever virus Antigen" patented technology

The invention relates to colloidal gold immunochromatographic test paper detecting African swine fever virus antigen, a preparation method and application thereof, belonging to virus disease diagnosistechnologies and the field of animal quarantine. The test paper comprises a PVC bottom board, wherein a sample pad, a colloidal gold pad, a nitrocellulose membrane and a water absorption pad are fixed on the PVC bottom board in sequence, the colloidal gold pad is internally provided with one epitope of monoclonal antibody Mab1 of an anti-ASFV p30 antigen marked by colloidal gold, the surface of the nitrocellulose membrane is provided with a detection line and a quality control line, the detection line is designed for another epitope of monoclonal antibody Mab2 for ASFVp30 antigen, and the quality control line is a goat anti-mouse IgG antibody. The colloidal gold immunochromatographic test paper has the advantages of being convenient, rapid, sensitive and specific, can detect ASFV infectedcells directly, and is suitable for ASFV infected diagnosis, epidemiologic investigation and international trade quarantine for swine.

The invention discloses an epitope antigen polypeptide of an African swine fever virus and an application of the epitope antigen polypeptide of the African swine fever virus. The epitope antigen polypeptide of the African swine fever virus is a polypeptide as shown in a sequence 2 or a polypeptide as shown in a sequence 3 in a sequence table. A composition of the epitope antigen polypeptide is thecomposition formed by the polypeptide as shown in the sequence 2 in the sequence table and the polypeptide as shown in the sequence 3 in the sequence table. According to the epitope antigen polypeptide or the composition thereof disclosed by the invention, a detection kit is prepared by coating an elisa plate with a chemically synthesized antigen peptide, so that the epitope antigen polypeptide is small in amount of antigen, high in sensitivity and high in specificity; whether an African swine fever virus antibody exists or not can be efficiently detected; and the kit has the advantages of being high in sensitivity, good in specificity and convenient to operate, and has a good market prospect

The invention discloses an African swine fever virus antigen duplex detection reagent and a preparation method thereof. The African swine fever virus antigen duplex detection reagent comprises a PVC bottom plate, wherein a sample pad, a colloidal gold marking pad, a nitrocellulose membrane detection pad and a water absorption pad are arranged at the top of the PVC bottom plate from left to right in sequence. According to the African swine fever virus antigen duplex detection reagent disclosed by the invention, by means of the detection of main and fairly conserved antigenic two proteins (p30 and p72) of AFSV, two results can be simultaneously obtained by once sample injection, furthermore, AFSV infection can be prompted as long as any one of the two results is positive, so that the detection rate is improved, whether early pig fever is caused by African swine fever virus infection can be quickly detected so as to discover the epidemic situation and take effective prevention and controlmeasures, furthermore, the African swine fever virus antigen duplex detection reagent has high sensitivity, high specificity, high speed (5-10min) ), simple and convenient operation, good repeatability, stable structure, good use performance, and is suitable for farmers to carry out self-examination or for professional institutions to carry out investigation of epidemic situation.

The invention discloses a kit and detection method capable of synchronously detecting African swine fever virus antigens and antibodies in swine blood, and belongs to the technical field of biology. According to the kit provided by the invention, a detection line for detecting African swine fever virus antigens and antibodies is arranged on an analysis membrane of a colloidal gold duplex card at the same time; preferably, the African swine fever virus P30 protein is used as an antibody for detection, the monoclonal antibody of the African swine fever virus P72 protein is used as an antigen fordetection, and the matched sample diluent is matched for use, so the African swine fever virus P30 protein antibody and the P72 protein antigen in the whole blood of the pig can be simultaneously detected through one-step operation.

The invention provides an African swine fever virus chimeric protein. The chimeric protein comprises: (1) an African swine fever virus p72 structural domain I; (2) an African swine fever virus p72 structural domain II; (3) an African swine fever virus p72 structural domain III; and (4) an African swine fever virus antigenic protein. According to the chimeric protein provided by the invention, the African swine fever virus p72 protein is used as a skeleton for the first time, antigenic epitopes of African swine fever virus antigenic proteins p54, p30, CD2v and p12 are well displayed, the immune effect is good, and remarkable body fluid and cellular immunity can be generated.

The invention discloses a eukaryotic-expression African swine fever virus p72 antigen and application thereof. The invention further discloses a detection kit comprising a p72 protein. The kit disclosed by the invention can be used for specifically detecting ASFV, is high in sensitivity and good in specificity and provides a novel detecting means for clinical diagnosis and epidemiological survey on ASFV infection.

The invention discloses a recombinant baculovirus for displaying African swine fever virus antigen P30 protein on the surface as well as a preparation method and application of the recombinant baculovirus. A gene sequence of P30 is inserted between an N-terminal signal peptide SP sequence of baculovirus envelope glycoprotein gp64 and a fusion structural domain TMD to construct a fusion gene gp64-Fp30, then the fusion gene gp64-Fp30 is connected with a recombinant vector pFPG to construct a recombinant expression vector pFPG-gp64-Fp30, then the pFPG-gp64-Fp30 is converted into DH10Bac cells to obtain recombinant Bacmid plasmids, finally, the recombinant Bacmid plasmids are transfected into Sf9 cells, and the recombinant baculovirus envelope glycoprotein gp64 fusion protein is obtained. The recombinant baculovirus with the surface displaying the African swine fever virus antigen P30 protein is obtained. The recombinant P30 protein produced by using the recombinant baculovirus has a conformation similar to that of natural protein, can be induced to generate a serum antibody with excellent sensitivity and specificity, can also realize efficient and rapid expression of the P30 antigen, and has a wide application prospect in preparation of ASFV diagnostic reagents or ASF vaccines.

The invention relates to a recombinant African swine fever antigen'cocktail 'vaccine containing an intramolecular adjuvant and application of the recombinant African swine fever antigen'cocktail' vaccine, and belongs to the field of biotechnological pharmacy, and the'cocktail 'vaccine contains three recombinant African swine fever virus antigen fusion proteins, namely OprI-p30-modified p54-TT, an N-segment amino acid sequence-TT of OprI-p72 epitope-pE248R and an N-segment amino acid sequence of OprI-CD2v and a C-segment amino acid sequence-TT of pEP153R. The invention provides expression and purification methods of the three recombinant African swine fever virus antigen fusion proteins, and component proportions and adjuvant compatibility for preparing the cocktail vaccine. The cocktail vaccine prepared by the invention can stimulate an organism to generate a neutralizing antibody and specific cellular immune response, and can be used as a subunit vaccine for preventing African swine fever.

The invention relates to a recombinant African swine fever virus antigen'cocktail 'vaccine and application thereof, and belongs to the field of biotechnological pharmacy, the'cocktail' vaccine is prepared by compatibility of three components including an N-segment amino acid sequence of recombinant protein p30-modified p54, p72 epitope-pE248R, an N-segment amino acid sequence of CD2v and a C-segment amino acid sequence of pEP153R with an adjuvant. The invention provides expression and purification methods of the three recombinant proteins, and proportions and adjuvants of the recombinant proteins for preparing the cocktail vaccine. The cocktail vaccine for immunizing pigs can stimulate the body to generate neutralizing antibodies and cellular immunity, and is an African swine fever subunit vaccine with a very good application prospect.

The invention provides polypeptides for promoting a swine body to generate an African swine fever virus (ASFV) antigen-specific immune response and application of the polypeptides, and belongs to thetechnical field of biological medicines. The polypeptides comprises one or more of a first polypeptide, a second polypeptide, a third polypeptide, a fourth polypeptide and a fifth polypeptide; the amino acid sequence of the first polypeptide, the amino acid sequence of the second polypeptide, the amino acid sequence of the third polypeptide, the amino acid sequence of the fourth polypeptide and the amino acid sequence of the fifth polypeptide are as shown by SEQ ID NO.1-SEQ ID NO.5 respectively. The polypeptides provided by the invention can significantly promote the proliferation of ASFV sensitized immune cells, and can promote the swine body to generate the ASFV antigen-specific immune response by promoting swine CD4+T and CD8+T lymphocytes to secrete interferon-gamma (IFN-gamma). Afteran animal is immunized, the immune response of the swine body can be remarkably promoted. The polypeptides or a polypeptide polymer obtained by polymerization of the polypeptides can be used for preparing subunit vaccines of the African swine fever virus.

The invention provides polypeptides for promoting a swine body to generate an African swine fever virus (ASFV) antigen-specific immune response and application of the polypeptides, and belongs to thetechnical field of biological medicines. The polypeptides comprises one or more of a first polypeptide, a second polypeptide, a third polypeptide, a fourth polypeptide and a fifth polypeptide; the amino acid sequence of the first polypeptide, the amino acid sequence of the second polypeptide, the amino acid sequence of the third polypeptide, the amino acid sequence of the fourth polypeptide and the amino acid sequence of the fifth polypeptide are as shown by SEQ ID NO.1-SEQ ID NO.5 respectively. The polypeptides provided by the invention can obviously promote the proliferation of ASFV sensitized immune cells, and can promote the swine body to generate the ASFV antigen-specific immune response by promoting swine mononuclear macrophages and B and T lymphocytes to secrete interferon-gamma (IFN-gamma). After an animal is immunized, the immune response of the swine pig body can be remarkably promoted. The polypeptides or a polypeptide polymer obtained by polymerization of the polypeptides can be used for preparing subunit vaccines of the African swine fever virus.

The invention discloses an antigen-binding protein, an antibody or an active fragment capable of specifically competing with a positive serum to bind to an African swine fever virus B646L antigen, an African swine fever virus competition ELISA detection kit comprising the antigen-binding protein, an antibody or an active fragment, and Its preparation method and application. The African swine fever virus competition ELISA detection kit of the invention has high sensitivity and good specificity, and can be used for detecting the specific antibody of the African swine fever virus. The competition ELISA detection kit can specifically compete for the antibody that binds to the B646L antigen in the serum, and has the advantages of rapidity, simplicity, sensitivity and specificity in the clinical detection of the African swine fever antibody.

The invention provides an African swine fever virus genetic engineering subunit oral vaccine, which comprises genetic engineering lactococcus lactis for recombinant expression of African swine fever virus antigen protein, and the antigen protein is ASFV p30 protein with an amino acid sequence of SEQ ID NO: 1, ASFV p54 protein with an amino acid sequence of SEQ ID NO: 3 or ASFV p72 protein with an amino acid sequence of SEQ ID NO: 5. The antigen protein is connected with an Escherichia coli heat-labile enterotoxin B subunit LTB gene protein. The African swine fever virus genetic engineering subunit oral vaccine can effectively stimulate local immune cells of intestinal tracts to generate secretory IgA, stimulate an organism to generate mucosal reaction, and further stimulate the organism to generate cellular immunity and humoral immunity through blood circulation.