39 results about "African Swine Fever Virus Infection" patented technology

The invention discloses a fluorescent quantitative PCR (Polymerase Chain Reaction) detection reagent, a kit and a detection method for an African swine fever virus (ASFV). The detection reagent is targeted to a conserved segment of an ASFV VP72 gene, and comprises a pair of specific primers, a specific probe and an internal labeling probe. The kit comprises a PCR mixed solution, Taq DNA polymerase, DEPC (diethylpyrocarbonate)-H2O, an ASFV positive quality product, an ASFV negative quality product, a quantification standard substance and a pseudovirus internal labeling solution, wherein the PCR mixed solution comprises a pair of specific primers, a specific probe and an internal labeling probe. The detection method comprises the steps of extraction of total DNAs, preparation of reaction components, making of a standard curve by diluting the standard substance, amplifying and result judgment. The reagent and the kit have the advantages of high specificity, high sensitivity, no pollution and high flux, and can be used for accurately detecting infection, inapparent infection or continuous carrying of a toxic host of low-content ASFV. The detection method is quick, specific and sensitive, and can be used for quantitatively detecting a large quantity of samples at the same time.

The invention discloses an RPA (recombinase polymerase amplification) primer and a detection kit for rapidly detecting African swine fever viruses. Target genes can be effectively amplified, specificity is 100%, detection sensitivity is 102 copy/reaction, and sensitivity is equivalent to that of fluorescent quantitative PCR (polymerase chain reaction). Cross reaction between the RPA amplificationprimer and one of classical swine fever viruses, vesicular exanthema swine viruses I, porcine reproductive and respiratory syndrome viruses, porcine circoviruses and the like is omitted. A RPA isothermal amplification system is rapidness in reaction and wide in temperature range, effective amplification of the target genes can be achieved at the temperature of 38-46 DEG C, and the detection kit can rapidly, efficiently and sensitively detect the African swine fever viruses and has the advantages that the kit is simple to operate, high in specificity, safe and free from pollution, reaction results are easily observed and the like. Effective technical means are provided for on-site rapidness detecting and screening of infection nucleic acid of the African swine fever viruses, and the RPA amplification primer has great significance for control of infection spreading of the African swine fever viruses in China and inspection and quarantine in infected areas and entry and exit ports.

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 provides a fluorescent quantitative PCR detection reagent, detection kit and detection method for identifying and diagnosing infection and immunization of African swine fever, and belongs to the technical field of biology. Specific primers and probes are designed aiming at CD2v of the African swine fever and eGFP of a fluorescent protein. An ASFVP72/CD2v dual fluorescent PCR detection method or ASFVP72/CD2v/eGFP triple fluorescent PCR detection method formed through combination with a P72 gene can be used for detecting the infection of the African swine fever and identifying a wild strain of a African swine fever virus and a CD2v gene deletion type recombinant vaccine strain, and thus, infected animals and immunized animals can be differentiated. The fluorescent PCR detectionkit has the advantages of simplicity and convenience in operation, high sensitivity, strong specificity, good repeatability and the like, and an important monitoring measure is provided for epidemicsituation detection and disease prevention and control.

The invention provides a PCR primer and probe for detecting infectivity of an African swine fever virus, a kit and a detection method. Primer sequences are shown in SEQ ID No. 1~2, and a probe sequence is shown in SEQ ID No. 3. The kit provided by the invention contains the PCR primer and probe and a nucleic acid dye, which is bonded to nucleic acid and has photosensitive characteristics; and whether the infective African swine fever virus is present in a sample to be detected or not is rapidly judged through pretreating the sample to be detected with the photosensitive nucleic acid dye, taking a sample free of photosensitive dye pretreatment as a control sample, then, extracting nucleic acids from the sample to be detected and the control sample, detecting African swine fever virus conserved gene copy numbers of the sample to be detected and the control sample separately by a qPCR method, and comparing the gene copy numbers of the sample to be detected and the control sample. The kitis short in detection consumed time, high in specificity, reliable in result and simple in result judgment, is applicable to any laboratory, prevention and control units and veterinary stations of alllevels of grass roots, large-, medium- and small-size farms and the like and is of far reaching importance in judgment on measures such as killing, prevention and control on the African swine fever virus.

The invention relates to a real-time fluorescent PCR amplification primer group for identifying and testing African swine fever viruses, the amplification primer group consists of upstream and downstream primers and probe primers of an African swine fever virus p72 gene, a CD2v gene and an MGF360-14L gene, and the invention also relates to a kit containing the real-time fluorescent PCR amplification primer group for identifying and testing the African swine fever viruses. By utilizing the high sensitivity of the primer group, wild strain infection or gene deletion vaccine strain immunization can be distinguished, the African swine fever virus infection can be prevented and controlled as soon as possible, and meanwhile, the kit can accurately detect suspected infection samples from different regional sources.

The invention discloses an immortalized porcine macrophage cell strain as well as a construction method and an application thereof, and belongs to the technical field of bioengineering. The immortalized porcine macrophage cell strain provided by the invention is obtained by co-transfecting immortalized genes SV40LT and hTERT into primary porcine bone marrow-derived macrophages in a lentivirus infection manner, the provided immortalized porcine macrophage cell strain is suitable for culturing African swine fever virus, can obtain high virus titer, is reliable and stable, can be continuously passed, and can be effectively applied to immunological research in the aspect of resisting African swine fever virus infection and production of African swine fever virus vaccines.

The invention belongs to the technical field of African swine fever treatment, and particularly relates to novel application of ML-60218 in preventing or treating African swine fever. It is accidentally finded that ML-60218 can remarkably inhibit the expression of African swine fever virus protein p30 and prevent viruses from invading host cells, and can be used for inhibiting early infection of ASFV; and the ML-60218 can obviously inhibit the replication of the African swine fever virus and reduce the virus titer after infection of the African swine fever virus, and can be used as an inhibitor of the African swine fever virus for preventing or treating the African swine fever.

The present invention discloses a class of shRNAs used for inhibiting replication of African swine fever viruses. The shRNAs contain two shRNA sequences such as SEQ ID NO.1 and SEQ ID NO.2. The present invention also provides a use of the aforementioned shRNAs in inhibiting the replication of the African swine fever viruses. The present invention also provides a recombinant expression vector containing the aforementioned shRNAs, and an adeno-associated virus (AAV) and a preparation method thereof. The provided shRNAs and recombinant AAV can be used to prepare drugs for preventing and/or treating African swine fever virus infection, such as therapeutic vaccines and can also effectively inhibit the replication and infection of the African swine fever viruses, and the preparation process is simple and conducive to large-scale production and promotion and application.

The invention discloses an African swine fever virus ELISA antibody detection kit, and belongs to the technical field of virus detection, the kit comprises an ELISA plate coated with antigens, and the antigens are prokaryotically expressed African swine fever virus p72, p30 and p54 proteins. The ELISA plate of the kit is coated with antigens, the antigens are prokaryotically expressed African swine fever virus p72, p30 and p54 proteins, the antigens are coated with extremely low antigen content, and the kit has no cross reaction with other main swine diseases and has good specificity through a specific test; the kit is high in sensitivity, good in repeatability and long in storage life, antibodies in different periods can be effectively detected through combined use of the three kinds of protein p72, p30 and p54 of the African swine fever viruses, a basis is provided for clinical swine herd infection conditions, and the kit plays an important role in preventing and controlling African swine fever virus infection.

The invention relates to an African swine fever virus p54 protein epitope peptide and application thereof. According to the invention, a new African swine fever virus p54 protein epitope is selected out through screening, the identification of the p54 protein N-terminal B cell epitope is completed, and the antigen epitope map of the p54 protein is improved. The selected antigen epitope (2DSEFFQPV9) is discovered for the first time, is highly conservative among various strains of African swine fever, and has important significance for serological detection of ASFV. The antigen epitope peptide and the antigen epitope peptide derived from the antigen epitope peptide and having the same function can be used for preparing a specific antibody of the African swine fever virus p54 protein, preparing a reagent for detecting or identifying the ASFVp54 protein, and laying a foundation for researching and developing vaccines for preventing and/or treating African swine fever virus infection.

The invention relates to the technical field of medicines, in particular to application of Triapine to treatment of African swine fever virus infection. The invention discloses application of Triapine and salt forms, hydrates, solvates and isotope substitutes thereof in preparation of drugs for treating African swine fever viruses. The invention has the following advantages: (1) Triapine has low toxicity to cells and is safe; and (2) Triapine has a good antiviral effect and can effectively treat African swine fever virus infection. (3) the selectivity coefficient is high, and safety is realized. And (4) the composition can be combined for use to enhance the treatment effect of African swine fever virus infection.

The invention discloses a method for rapidly detecting African swine fever virus by using a fluorescence in-situ detection technology. The method comprises the following steps: preparing a fluorescent probe, collecting and fixing cells of a sample to be detected, and hybridizing and dyeing the fluorescent probe and the cells, observing the dyed hybrid cells, and identifying whether the sample to be detected is infected by the African swine fever virus; signals are amplified through a primer pool method, a signal point is composed of thousands of DNA fragments, compared with a traditional in-situ hybridization method, the signals are stronger and easy to capture, a super-resolution imaging system is not needed, a signal cluster can be analyzed through a common fluorescence microscope, and the experiment cost and the operation difficulty are reduced.

The invention discloses application of a small molecule compound in anti-African swine fever virus ( ASFV) infection, and belongs to the technical field of medicine. According to the invention, an enzyme active center of E165R to exert hydrolytic action is defined by carrying out a crystal structure analysis of an E165R-dUMP complex, and then 1,2,3,4,6-O-Pentagalloylglucose capable of matching theE165R enzyme active center and significantly inhibiting E165R enzyme activity is obtained through in vitro screening by using the enzyme active center of E165R as a target. Further virus level experiments show that the 1,2,3,4,6-O-Pentagalloylglucose can significantly inhibit the replication of the ASFV in porcine alveolar macrophages, with high application value in clinical treatment or prevention of ASFV infection.

The invention belongs to the technical field of biology, and discloses an African swine fever virus CD2v truncated protein as well as application thereof in preparation of wild virus and natural attenuated virus detection kit Through comparison, it is found that positive serum infected by the African swine fever virus cannot be detected when prokaryotic expression CD2v truncated protein serves as the ELISA coating antigen; however, positive serum infected by the African swine fever virus can be detected when eukaryotic expression CD2v protein serves as the ELISA coating antigen. An indirect ELISA kit prepared by taking the eukaryotic expression CD2v protein as a coating antigen is matched with an indirect ELISA kit taking the African swine fever virus p54 protein as a coating antigen, so that the kit can be used for identifying infection of ASFV wild strains and natural attenuated strains; and moreover, the operation steps are simple, and the result is reliable. Therefore, the African swine fever virus indirect ELISA detection kit prepared from the eukaryotic expression truncated protein provided by the invention is very suitable for clinical large sample detection, and is suitable for large-scale popularization.

The invention relates to the technical field of medicines, in particular to application of Tilorone to prevention/treatment of African swine fever virus infection. The invention relates to application of telolone and salt forms, hydrates, solvates and isotope substitutes thereof in preparation of drugs for preventing or treating African swine fever viruses. The invention has the following advantages: (1) the bioavailability is high, and the half-life period is long; and (2) the maximum tolerance dose is high, the selectivity coefficient is high, and the safety is high. And (3) the activity of preventing/resisting African swine fever virus infection is strong. And (4) the drug delivery modes are multiple, and the application range is wide. And (5) the composition can be combined for use and has a good effect of preventing/treating African swine fever virus infection.