426 results about "Classical swine fever virus CSFV" patented technology

The invention discloses a four-gene-deletion weak-toxin strain for African swine fever viruses. The weak-toxin strain is the four-gene-deletion weak-toxin strain for an African swine fever virus SY18separation strain, and the following gene function protein is deleted: CD2v gene coding products and three multigene family genes( MGF360-12L, MGF360-13L and MGF360-14L ) coding products. The invention further discloses an application of the weak-toxin strain of the African swine fever viruses to preparation of vaccines for preventing or treating African swine fever. The weak-toxin strain of the African swine fever viruses can provide complete immunoprotection effect on attack of ASFV parent toxin strains, is high in safety, and is suitable for being used as vaccine candidate strains for preventing the African swine fever.

The invention discloses a colloidal gold immunochromatographic test strip for detecting wild-type classical swine fever virus, which consists of water absorbent paper (1), a cellulose nitrate membrane (2), a colloidal gold pad (3), a sample pad (4) and a support (5), wherein the cellulose nitrate membrane contains a detection line which is formed by coating monoclonal antibody HQ06 of anti-classical swine fever virus E2 protein and a quality control line which is formed by coating rabbit anti-mouse IgG antibody; and the colloidal gold pad is combined with colloidal gold-labeled monoclonal antibody 6E10 of the anti-classical swine fever virus E2 protein. The test strip does not react with C-strain of classical swine fever virus, bovine viral diarrhea virus, porcine reproductive and respiratory syndrome virus, transmissible gastroenteritis virus, porcine epidemic diarrhea virus, porcine rotavirus, pseudorabies virus, porcine parvovirus and porcine circovirus type 2, and can accurately and sensitively identify the wild-type classical swine fever virus, thereby having good specificity, sensitivity and repeatability.

The invention relates to an ELISA kit for distinctively detecting antibodies of classical swine fever (CSF) vaccine immunity and wild virus infection and a preparation method thereof. An indirect ELISA kit or a blockage ELISA kit is formed by expressing and purifying classical swine fever virus (CSFV) non-structural protein NS3, coating a solid-phase carrier and assembling with other matched reagents. The kit has the characteristic of distinctively detecting different antibodies generated by the CSF vaccine immunity and the wild virus infection. The kit can distinctively diagnose the CSF vaccine immunity and the wild virus infection.

The invention discloses a recombinase polymerase application (RPA) primer for rapidly detecting African swine fever virus (ASFV) nucleic acid, a preparation method of the RPA primer, and a kit, belonging to the technical field of biology. A pair of RPA primers, i.e., an upstream primer <210>2 and a downstream primer <210>3 which have high specificity and strong sensibility are screened out; an RPAdetection system for rapidly detecting the ASFV nucleic acid is further established by means of the primers. Compared with the common polymerase chain reaction (PCR) method, RPA-lateral flow assay (LFA) has the advantages that firstly, the RPA-LFA belongs to an isothermal amplification technology and is low in requirements for instruments and equipment, and reactions can be completed only by means of a constant temperature water bath kettle; secondly, the detection speed of the RPA-LFA is fast, and the reaction time is 40 minutes and is shorter than the conventional PCR reaction time; thirdly, the visualization of detection results can be realized. Due to the characteristics, the RPA-LFA method established by the invention can be used for rapid detection and differential diagnosis of ASFVin common laboratories of grassroots units.

The invention relates to an African swine fever virus nucleic acid amplification primer, a detection method and a kit. The primer pair and the specific nucleic acid probe are selected from SEQ ID NO: 1-12. The detection method comprises the following steps: (a) amplifying nucleic acid in the sample to be detected by using a PCR primer; (b) labeling the detection probe labeled by alkyl sulfydryl group on nano gold particles; (c) hybridizing the capture probe labeled by biotin and the detection probe labeled by nano gold in step (b) with a metamorphic PCR product; (d) adding the hybrid system in step (c) to a Streptavidin-coated ELISA plate to capture the hybrid compound; (e) carrying out silver enhancement to capture nano gold; (f) and stoping the silver enhancement reaction, and visually inspecting the grey scale judgment result. The invention has the characteristics of high detection sensitivity, strong detection specificity and low detection cost, can effectively eliminate false positive or false negative result when the PCR method is used for detecting African swine fever virus, and quickly detect the African swine fever virus nucleic acid.

The invention discloses a triple fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection reagent for African swine fever viruses, swine fever viruses and respiratory syndrome viruses and a preparation method and application thereof. Three sets of specific primers and Taqman probes as well as positive controls specific to African swine fever virus CP530R genes, swine fever virus 5 minute-UTR genes and swine respiratory syndrome virus NSP2 genes respectively are designed and synthesized, and a rapid, easy and convenient triple fluorescent RT-PCR detection system with high specificity and high sensitivity is established by using the three sets of primers and probes, so that nucleic acids of the African swine fever viruses, the swine fever viruses and the respiratory syndrome viruses can be detected synchronously from a detected sample within 3-4 hours in a rapid, accurate, specific, safe, easy and convenient way. The detection reagent can be applied to synchronous detection of the nucleic acids of trace African swine fever viruses, swine fever viruses and respiratory syndrome viruses in hogs and relevant samples thereof.

The invention discloses a monoclonal antibody against virulent strain E2 protein of classical swine fever virus and a hybridoma cell strain secreting the monoclonal antibody. The hybridoma cell strain is obtained by using hog cholera lapinized virus vaccine strain E2 protein expressed by Baculovirus as tolerogen, selecting Shimen strain E2 protein as immunogen, immunizing mouse by cyclophosphamide immunosuppression method, carrying out cell fusion, and sieving hybridoma cell strain capable of stably secreting monoclonal antibody against E2 protein. The monoclonal antibody can react with Shimen strain and can produce specific reaction with virulent strain of classical swine fever viruses of 1.1, 2.1, 2.2 and 2.3 gene sub-groups. The monoclonal antibody has neutralization activity and does not react with hog cholera lapinized virus vaccine strain, so that the monoclonal antibody can be used for differentiating virulent strain of classical swine fever virus and hog cholera lapinized virus vaccine strain, which establishes the foundation for establishing a method for differentiating wild virus infection of classical swine fever and vaccine immunity and for researching the molecular difference between CSFV virulent strain and mild strain.

The invention provides a triple real-time fluorescent quantitative PCR detection primer for detecting an African swine fever wild strain and a gene deletion strain, and a kit and a detection method thereof. The triple fluorescent quantitative detection kit is developed and researched for three genes CD2V, VP72 and MGF-360 14L of the African swine fever virus by utilizing a multiple fluorescent PCRtest means, and whether a sample is infected with the African swine fever virus and whether gene deletion exists in the infected virus or not can be determined at the same time. The method can detecta large number of samples at the same time, provides an effective tool for scientifically and reasonably preventing and controlling African swine fever, guarantees the healthy development of the pigindustry, and has the advantages of convenience in operation, high sensitivity, strong specificity, short detection time and the like.

The invention discloses a dual fluorescence PCR detection reagent, a kit and a detection method for swine fever virus and African swine fever virus, and belongs to the field of animal pathogen detection. Aiming at a 5'-UTR gene of the swine fever virus and a P72 gene of the African swine fever virus, primers and probes capable of covering all strains and suitable for double detection are separately designed and screened, including two pairs of specific primers and two specific probes; the invention also describes a kit containing the primers and the probes and a PCR detection method using theprimers and the probes; the invention, through the design of the primers and the adjustment of each component of PCR, realizes purposes of one-time analysis and simultaneous detection and differentiation of the swine fever virus and the African swine fever virus on the premise of no reduction in sensitivity and specificity, which not only reduces the workload and cost of detection, but also greatly saves the detection time, thus gaining valuable time for epidemic disease prevention and treatment.

The invention relates to a condon optimized African swine fever virus P54 gene and a nucleic acid vaccine based on the gene, wherein the nucleic acid vaccine consists of an eukaryotic expression vector and the condon optimized African swine fever virus P54 gene. The nucleic acid vaccine is constructed through the following steps of optimizing the condon of P54protein, designing a specific primer, transferring sequence of the artificially synthesized African swine fever virus P54 gene into a cloning vector and recombining the purified P54 gene into the eukaryotic expression vector. The nucleic acid vaccine can be used for preventing the occurrence of African swine fever and is a powerful tool for solving the immune prevention and control of the African swine fever. Compared with the traditional vaccines, the nucleic acid vaccine has the advantages of small quantity, security, long-term effect, stability and convenience in operation when reaching same immune effect.

The invention relates to an African swine fever virus ASFV-LAMP detection primer set, reagent kit and detection method. In order to cope with the epidemic situation of African swine fever in Chin at 2018, an African swine fever virus P72 gene is used as an LAMP detection primer set. The primer set specially detects the African swine fever virus gene P72, ASFV can be effectively detected, a new technique means is provided for prevention and control of the African swine fever, and toxin strain detection and entry and exit quick screening are facilitated. The LAMP detection method can realize quick detection of the African swine fever viruses, detection results can be directly tested visually, and the entire reaction can be realized only needing heating. The relying of a traditional nucleic acid detection technique on a PCR instrument can be shaken off. The method is high in detection sensitivity, high in specificity, good in repeatability and high in detection speed, and can be used as an effectively on-field detection means for African swine fever.

The invention provides a preparation method of a recombined new castle disease virus vaccine strain which expresses African swine fever virus p72 proteins and a recombined new castle disease virus vaccine strain. The method provided by the invention comprises the following steps: constructing a recombinant transcriptional plasmid which is inserted with a p72 gene of African swine fever virus (ASFV); constructing a transcriptional helper plasmid system; carrying out a contransfection for the transcriptional plasmids and the transcriptional helper plasmids into host cells BHK-21 which can be duplicated in new castle disease attenuate strains; and saving and obtaining the recombined virus stain. The vaccine strain for expressing the African swine fever virus p72 proteins provided by the invention has important preservation and strategy meaning in the aspect of animal epidemic disease prevention and control, and can be applied to the treatment and prevention of African swine fever virus.

The invention discloses a method for identifying a wild strain and a vaccine strain of a hog cholera virus. According to the method, a primer specially used for identifying the wild strain and the vaccine strain of the hog cholera virus is designed in a conservative region shared by the wild strain and the vaccine strain. The primer is high in specificity and can well multiply the wild strain and the vaccine low virulent strain; according to a reverse transcription-polymerase chain reaction technology and a high resolution melting (HRM) technology, the wild strain and the vaccine strain can be obviously identified. The method is a simple, quick and practical novel technology for identifying the wild strain and the vaccine strain of the hog cholera virus.

The invention provides a pig virus gene chip and a detection method thereof. The gene chip comprises a probe fixedly arranged on a substrate carrier, wherein the probe is selected from the characteristic segments of the following viruses: a classical swine fever virus (CSFV), a porcine reproductive and respiratory comprehensive virus (PRRSV), a pseudorabies virus (PRV), a porcine circovirus (PCV) and a porcine parvovirus (PPV). The types of the viruses detected by the method are various and the viruses cover common porcine viruses substantially. Random primer polymerase chain reaction (PCR) and multiplex-PCR are adopted to mark, so that false positive which is easy to cause when a PCR result is detected by gel electrophoresis is avoided, and high-flux accurate detection with short time is realized. The substrate carrier is a glass sheet which is subjected to aldehyde treatment, so that the combination between the probe and a target is facilitated, and higher noise is not brought to detection.

The invention discloses a molecular probe for detecting African swine fever virus, a kit and application thereof, so as to solve the technical problems of poor reliability and poor practicability of the existing detection technology. The nucleotide sequence of the molecular probe for detecting African swine fever virus is shown as in SEQ ID NO.2. An RT-PCR reaction solution for detecting African swine fever virus is prepared, lyophilized powder for the RT-PCR reaction solution for detecting African swine fever virus is prepared, an African swine fever virus detection kit is prepared, and an African swine fever virus qualitative and quantitative detection method is provided. The designed molecular probe and the built RT-PCR detection method have the characteristics of high sensitivity, goodrepeatability and strong specificity, the reliability and the usability of ASFV detection by the PCR technology are improved, pollution-free and high-flux detection is realized, and ASFV nucleic acids in susceptible animals and related animal products can be detected.

The invention relates to the technical field of livestock immunology and antibody preparation and especially relates to an anti-African swine fever antibody and a preparation method thereof. The antibody is prepared according to the following steps: using an African swine fever virus for immunizing a healthy female poultry animal; collecting yolks after the content of African swine fever antibodyin the yolks of the immunized poultry animal exceeds 40ng/mL; extracting water-soluble components in the yolks; purifying, thereby acquiring the antibody. The preparation technology for producing theantibody is simple, low-cost and high-feasibility and can be used for large-scale preparation of African swine fever antibody. The acquired antibody can be applied to contaminated pig farms and earlyinfected swine, so as to reduce the duplication and propagation of African swine fever virus.