145 results about "Wild strain" patented technology

The invention discloses a kitchen waste oil-based sophorolipid high-yield candida utilis strain and an application thereof. Belongs to the technical field of directional domestication breeding. According to the method, wild candida bombicola is used as an original strain, and is sequentially domesticated in a culture medium containing kitchen waste oil with the concentration gradient of 2%, 4%, 6%, 8% and 10% to obtain the S.bombicola x10. The invention further discloses an application of the domesticated strain in fermentation production of sophorolipid, after the strain is subjected to shake flask fermentation, the sophorolipid yield reaches 60.19 g / L and is improved by 16.22% compared with that of a wild strain, and the acid type sophorolipid yield is improved by 32.11%. When the concentration of the lactone sophorolipid generated by fermentation is 1.25 mg / mL, the growth of the bacillus subtilis is completely inhibited. The invention provides an excellent production strain for industrial production of sophorolipid, and has good theoretical research significance and practical application value.

The invention discloses a streptococcus suis type 2 gene deletion strain as well as a construction method and application thereof, and belongs to the technical field of biological products. According to the invention, the B9H0105210 gene in the genome of the streptococcus suis serotype 2 SC19 is deleted in a homologous recombination manner for the first time, the hydrogen peroxide and nitric oxide stress resistance of the constructed streptococcus suis serotype 2 gene deleted strain is reduced, and the pathogenicity to mice is obviously weakened; moreover, the growth performance of the streptococcus suis type 2 gene-deleted strain is not greatly different from that of a wild strain SC19, the condition of gene recovery does not exist after continuous passage, and the streptococcus suis type 2 gene-deleted strain has an application prospect as an attenuated live vaccine, has a good immune protection effect on different genotypes of streptococcus suis such as the streptococcus suis type 2 strain SC19 and streptococcus suis type 2 P1 / 7, and has a good application prospect. And a certain cross immune protection performance is achieved.

The invention discloses an E3 ubiquitin ligase gene ulA, the nucleotide sequence of which is shown as SEQ ID NO: 1. The gene is knocked out from fusarium oxysporum by using a homologous recombination method to obtain a mutant strain ulA; compared with a wild strain, the mutant hulA has the advantages that the spore yield is reduced, the spore germination is delayed, the response capability to various abiotic stresses is reduced, the pathogenicity to panax notoginseng plants is reduced, and the sensitivity to six bactericides is increased, and experimental results show that the gene plays an important role in pathogenicity, drug resistance and the like of fusarium oxysporum. The gene can be used as a target gene for developing an anti-fusarium oxysporum preparation.

The invention relates to the technical field of carotenoid cleavage dioxygenase, and discloses a carotenoid cleavage dioxygenase mutant and application thereof. In order to improve the efficiency of converting carotenoid in tobacco into aroma molecule beta-ionone, specific mutation of characteristic sites is carried out on carotenoid cleavage dioxygenase (PhCCD1) from petunia, that is, serine (S) at the 428 site of the carotenoid cleavage dioxygenase is mutated into cysteine (C) or tyrosine (Y) to obtain a mutant S428C or S428Y, and the mutant S428C or S428Y is converted into beta-ionone. The yield of the beta-ionone converted from the carotenoid is greatly improved. Experiments prove that the yields of beta-ionone of S428C and S428Y respectively reach 54.2 mg / L and 50.6 mg / L, which are respectively increased by 69.7% and 58.5% compared with those of a wild strain WT.

The invention belongs to the technical field of microorganisms, and particularly relates to stenotrophomonas maltophilia yg3 and application thereof. The invention discloses stenotrophomonas maltophilia yg3, and the preservation number of the stenotrophomonas maltophilia yg3 is CGMCC (China General Microbiological Culture Collection Center) No.34131. The stenotrophomonas maltophilia yg3 disclosed by the invention has the capability of efficiently synthesizing indoleacetic acid (IAA), and the IAA yield of 98.10 mg / L can be realized without optimization in an NB culture medium containing 5mM L-tryptophan, which is obviously higher than that of other wild strains. Moreover, the stenotrophomonas maltophilia yg3 has a remarkable growth promoting effect on plants (such as hot peppers), pot experiments show that the stenotrophomonas maltophilia yg3 can remarkably increase the number of effective leaves, the plant height and the biomass of the hot peppers, and the stenotrophomonas maltophilia yg3 can be applied to the field of microorganisms. Particularly, the method has important application value in the fields of microbial plant growth promoters and green synthesis of IAA (indoleacetic acid).

The invention discloses engineering chlamydomonas reinhardtii for synthesizing high-value pigment and a construction method and application thereof.An engineering chlamydomonas reinhardtii strain Blup-BKT-CHYB-CHLC is preserved in the China General Microbiological Culture Collection Center on September 1, 2025, and the preservation number is CGMCC NO.46634. The content of astaxanthin in the constructed engineering chlamydomonas reinhardtii strain reaches 25.5 mg / L, and the content of astaxanthin in the engineering chlamydomonas reinhardtii strain reaches 9.5 mg / L; in addition, the light energy utilization efficiency is remarkably enhanced, the stress resistance is improved, very strong oxidation resistance is achieved, the accumulation of astaxanthin endows the engineering strain with highlight tolerance, the yield of high-value pigments is increased, and the biomass of the engineering strain is greatly superior to that of a wild strain under the highlight condition. In addition, the chlorophyll c synthase is applied to the reconstruction of a pigment system of the chlamydomonas reinhardtii for the first time, a frontier tool is quickly converted, and a molecular mechanism of'chlorophyll c-astaxanthin-blue pigment 'for synergistically regulating the photosynthetic efficiency of the chlamydomonas reinhardtii and product synthesis is clarified for the first time. The research blank of pigment system coupling regulation and control of photosynthesis and metabolism in the chlamydomonas reinhardtii is filled.

The invention discloses a construction method of a nocardia seriolae mycolic acid deficient strain, which belongs to the technical field of aquaculture prevention and control, and comprises the following steps: strain and material preparation, culture and genome extraction, upstream and downstream fragment cloning to construct deletion plasmids, competent cell preparation, electrotransformation and screening, and biological characteristic and toxicity analysis. The nocardia seriola seriola mycolic acid defect type strain is constructed and compared with a wild strain and a back-filling strain for research, the effect of the mycolic acid in the aspects of growth, form, toxicity and the like of thalli can be defined, the relationship between the mycolic acid and the pathogenicity of the thalli can be deeply known, and an important clue is provided for revealing the pathogenic mechanism of the nocardia seriola seriola.

The invention relates to a monoclonal antibody DIDA55 aiming at hog cholera virus, and a preparation method and application thereof, belonging to the field of medical preparations. The monoclonal antibody DIDA55 or an antigen binding fragment thereof comprises a heavy chain variable region and a light chain variable region, amino acid sequences of LCDR1, LCDR2 and LCDR3 in a light chain variable region of the antibody are shown as 24th to 34th, 50th to 56th and 89th to 97th in SEQ ID No: 4; amino acid sequences of HCDR1, HCDR2 and HCDR3 in a heavy chain variable region of the antibody are shown as the 31st to 35th, the 50th to 65th and the 95th to 102th of SEQ ID No: 5. The monoclonal antibody provided by the invention can be used in the fields of swine fever E2 protein labeled subunit vaccine immunity, serological diagnosis of wild strains, swine fever vaccine immune effect evaluation, related experiments and the like, and provides antibody resources for prevention, control and purification of CSF.

The invention belongs to the technical field of biology, and particularly relates to a human metapneumovirus mouse adapted strain and application thereof. Compared with a wild strain, the strain has higher susceptibility to animals; the strain can be used for treating BALB / c mice and can cause typical clinical symptoms and even death, for example, after the strain infects the BALB / c mice, the disease symptoms are obvious, the weight of 83% infected mice is severely reduced to 75% or below and reaches the humanity killing standard, and the death rate reaches 50% or above 4 days after the strain infects the BALB / c mice; the strain can be used for constructing a cell model or an animal model of human metapneumovirus infection or diseases caused by the human metapneumovirus infection, and is further used for screening drugs for preventing and / or treating the human metapneumovirus infection or the diseases caused by the human metapneumovirus infection. In addition, the strain can also be used for preparing medicines for preventing and / or treating human metapneumovirus infection or diseases caused by the human metapneumovirus infection, preparing antibodies or antiserum for resisting the human metapneumovirus and the like.

The invention discloses an application of an isoeugenol monooxygenase mutant in production of isoeugenol vanillin, which comprises the steps of recombinant strain construction, fermentation culture of recombinant strains and catalytic reaction of fermentation broth, and the isoeugenol monooxygenase mutant is mutated through an isoeugenol monooxygenase amino acid sequence of wild fungi WT. And the thalli are expressed by escherichia coli, so that a series of thalli with a promoting effect on the catalytic efficiency of vanillin production are formed. The mutant super-efficient enzyme catalyst with the catalytic conversion rate reaching up to 98.5% or above is obtained through screening, the defects of low efficiency, pollution, high energy consumption and the like of traditional chemical catalyst production are overcome, and the mutant super-efficient enzyme catalyst is more suitable for green industrial production requirements and worthy of being popularized to industrial use.

The invention discloses kluyveromyces marxianus and application of the kluyveromyces marxianus in production of single-cell protein by taking straw as a raw material. The Kluyveromyces marxianus is a ZH-2 bacterial strain which is CGMCC (China General Microbiological Culture Collection Center) No.32295 The strain is obtained from a wild strain in a laboratory adaptive evolution mode, wheat straw and an inorganic nitrogen source can be used as raw materials for efficiently producing single-cell protein, compared with a parent strain, the utilization speed and the utilization rate of biomass, glucose and xylose are remarkably increased, the yield of ZH-2 SCP can reach 0.047 g.g <-1 >, and the yield of ZH-2 SCP can reach 0.045 g.g <-1 >. And the essential amino acid mode basically accords with adult and infant amino acid modes, and can be used as high-quality protein nutrition supply. The Kluyveromyces marxianus ZH-2 strain capable of obtaining single-cell protein by fermenting wheat straw as a raw material is provided from the perspective of utilizing cheap waste resources, so that the production cost of SCP is greatly reduced, and the problem of protein resource shortage is effectively solved.

Provided are an Aeromonas dhakensis mutant strain with deletion of a surface polysaccharide synthesis gene, a complemented strain, a construction method therefor, and the use thereof. The mutant strain is obtained by deleting one or more of ugd, ugd2, and cap1J from the Aeromonas dhakensis wild strain A. dhakensis C160501.

The invention relates to the field of microorganisms, in particular to bacillus velezensis and application of the bacillus velezensis in preparation of surfactin, the preservation number is CCTCC NO: M 20251689, the bacillus velezensis is a wild type bacillus velezensis strain, and the surfactin yield of the bacillus velezensis is 2-3 times higher than that of a conventional wild strain. At the shake flask level, the yield of the surfactin reaches 10.4 g / L, when fed-batch fermentation is carried out in a 5L fermentation tank, the yield reaches 22.3 g / L, the yield and production intensity are higher than the currently known wild fungus fermentation level, and the method has huge industrial application potential.

The invention discloses a schizochytrium limacinum strain for efficiently converting kitchen waste to produce single-cell protein and application of the schizochytrium limacinum strain, and belongs to the technical field of biological engineering. The preservation number of the schizochytrium sp LOX-2 is CGMCC (China General Microbiological Culture Collection Center) No. 41623. The invention also discloses a method for producing single-cell protein by efficiently converting kitchen waste. The method comprises the following steps: fermenting food waste to produce volatile fatty acid; and culturing schizochytrium limacinum by using the strain as a carbon source to obtain single-cell protein. According to the recombinant strain spLOX-2, the utilization of lactic acid is improved, the biomass and the protein yield of the recombinant strain spLOX-2 are respectively improved by 14.19% and 12% compared with those of a wild strain, and the grease content is obviously improved by 107.1%. According to the invention, a new way is provided for a sustainable biological economic mode, wastes are converted into valuable biological chemicals, and a solid foundation is laid for expanding the scale of the process and the economic feasibility of the process in industrial application.

The invention provides a nucleic acid probe, a primer group, a kit, a detection method and application, and relates to the technical field of biology. The nucleotide sequence of the nucleic acid probe is as shown in SEQ ID NO. 1; wherein the 3'end of the nucleic acid probe is modified with MGB, and locked nucleic acid is modified at a base position corresponding to the difference between the swine fever wild strain and the swine fever vaccine strain in the nucleotide sequence. According to the nucleic acid probe, a specific sequence is utilized to precisely target a swine fever wild strain, and locked nucleic acid modification is introduced at a differential site, so that the recognition capability on single-base mismatch is remarkably enhanced, non-specific binding to a vaccine strain is thoroughly blocked, and false positive interference is eliminated; and 3 '-terminal MGB modification is matched to improve the binding stability and sensitivity, so that the efficient and accurate identification of the swine fever wild strain under the immune background is ensured.

The invention belongs to the technical field of virus detection, and discloses a digital PCR (polymerase chain reaction) kit for simultaneously detecting African swine fever virus, porcine reproductive and respiratory syndrome virus and swine fever virus, and the kit comprises primers and probes as shown in SEQ ID NO.1-SEQ ID NO.24. After a single system of an African swine fever virus-VP72 gene, an African swine fever virus-CD2V gene, an African swine fever virus-14L gene, a porcine reproductive and respiratory syndrome virus-RdRP gene, a porcine reproductive and respiratory syndrome virus-European strain, a porcine reproductive and respiratory syndrome virus-American strain, a swine fever virus-vaccine strain and a swine fever virus-wild strain is tested, a nine-fold digital PCR system containing an internal standard is established; through tests of indexes such as linearity, specificity, minimum detection limit, repeatability, comparison of a single digital PCR system and a multiple digital PCR system, the currently developed digital PCR multiple detection kit completely meets development requirements and is high in specificity and sensitivity, and the minimum detection limit of the digital PCR multiple detection kit can reach 10 Copies / mu L or below.

The present invention belongs to the field of selective culture of microorganisms and relates to a detection medium for Burkholderia cepacia complex in a pharmaceutical water system. A detection medium for Burkholderia cepacia complex in a pharmaceutical water system, the formula (g / L) of the detection medium is: peptone 0.5 - 2.0 g, sodium pyruvate 1 - 4 g, carbon source 1 - 4 g, yeast extract 0.5 - 2.0 g, casein amino acid 1 - 4 g, magnesium sulfate 0.05 - 0.2 g, and the solvent is water. The inventor has screened a detection medium for Burkholderia cepacia complex in a pharmaceutical water system with special components. Through the culture experiment of Burkholderia cepacia complex, it is found that whether it is a standard strain or a wild strain isolated from a pharmaceutical water system, within the same culture time, the OD 600 value results of 1 / 10 TSB and R2A liquid media do not exceed 0.5, while the bacterial concentration in the detection (BCB) medium for Burkholderia cepacia complex in a pharmaceutical water system is significantly higher than that in 1 / 10 TSB and R2A liquid media.

The invention relates to the technical field of agricultural biology, in particular to application of a regulatory gene Mt2312847 in improving the cellulase expression quantity of mycelia thermophila. Through interference or knockout of a regulatory gene Mt2312847 of a wild strain of the mycelia thermophila and knockout of the strain (Mt delta 2312847), the cellulase concentration of fermentation liquor of the interference strain is higher than that of the wild strain, and the CMCase enzyme activity of the Mt delta 2312847 and the CMCase enzyme activity of the Mt delta 2312847 are 2.2 times and 1.6 times that of a control strain.

The invention discloses a dual fluorescent quantitative PCR (polymerase chain reaction) primer probe combination for identifying a PRV (pseudorabies virus) vaccine strain and a wild strain. The dual fluorescent quantitative PCR primer probe combination comprises two pairs of primers gB-F and gB-R, gE-F and gE-R, and probes gB-probe and gE-probe, wherein the two pairs of primers are used for specifically amplifying a PRV gB gene and a PRV gE gene; through existence of real-time fluorescence PCR amplification curves of the PRV gB gene and the gE gene, a PRV vaccine strain (gE gene deletion) and a wild strain can be rapidly, specifically, sensitively and stably identified, technical support can be provided for porcine pseudorabies epidemic disease monitoring, and thorough purification of the PRV in a live pig breeding link is facilitated.

The invention discloses recombinant xanthomonas campestris with xanthan gum hydrolase displayed on the surface and a construction method and application of the recombinant xanthomonas campestris. The recombinant xanthomonas campestris for surface display of xanthan hydrolase is constructed by introducing a recombinant vector obtained by fusion expression of an ice crystal nuclein (INP) gene and a xanthan hydrolase gene into xanthomonas campestris. The gel yield of recombinant xanthomonas campestris fermentation is twice that of a wild strain and reaches 6.7 g / L. The strain can efficiently hydrolyze xanthan gum with high molecular weight (such as more than 100 wDa), the traditional production process which is relatively complicated is simplified, and a new method is provided for large-scale production of the xanthan gum with low molecular weight.

The invention relates to construction and application of an engineering bacterium for high yield of chitinase ChiA, in particular to a bacillus thuringiensis engineering bacterium HD73 delta cdsR (P5014-SPchiD-chiA) strain, and the preservation number of the bacillus thuringiensis engineering bacterium HD73 delta cdsR (P5014-SPchiD-chiA) strain is CGMCC (China General Microbiological Culture Collection Center) No.35677. The HD73 endogenous gene chiA gene is efficiently expressed in the bacillus thuringiensis delta cdsR, and the yield in a fermentation culture medium is far higher than that of a wild strain HD73. Compared with HD73 (P5014-SPchiD-chiA), the compound has a relatively strong inhibition effect on mycelial growth of fusarium graminearum and exserohilum turcicum, and shows a better effect on prevention and treatment of larvae of plutella xylostella compared with HD73 (P5014-SPchiD-chiA).

The invention relates to the technical field of rotaviruses, in particular to a G8P [8] type group A rotavirus CZ-23 and application thereof. The invention relates to a G8P [8] type group A rotavirus CZ-23, which is preserved in China General Microbiological Culture Collection Center (CGMCC) on April 11, 2023, and the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the preservation number is CGMCC (China General Microbiological Culture Collection Center) The invention provides a novel human rotavirus wild strain which is reported and separated for the first time in China, and the strain is strong in virus replication capability and good in hereditary stability, can be applied to development of vaccine strains of human rotavirus vaccines, and is suitable for attenuated rotavirus oral vaccines.

The application belongs to the technical field of biotechnology, and particularly relates to a human metapneumovirus mouse-adapted strain and application thereof. Compared with wild strains, the strain can exhibit stronger susceptibility to animals, and can cause typical clinical symptoms and even death. For example, after the strain infects BALB / c mice, the disease symptoms are obvious, 83% of the mice have a severe decrease in body weight to below 75% after infection, and have reached the humane killing standard. Four days after the strain infects BALB / c mice, the mortality rate is above 50%. The strain can be used to construct a cell model or an animal model of human metapneumovirus infection or a disease caused by the human metapneumovirus, and is further used to screen drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus. In addition, the strain can also be used to prepare drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus, and to prepare antibodies or antisera against the human metapneumovirus.

The present invention discloses the application of miRNA derived from insect vectors in inhibiting rice stripe virus infection, belonging to the field of biotechnology. The miRNA is the following miRNA A1) and A2): A1) a single-stranded RNA molecule having a nucleotide sequence of SEQ ID No.1; A2) a miRNA obtained by replacing and / or deleting and / or adding nucleotides of the nucleotide sequence shown in SEQ ID No.1, which has more than 90% identity with the RNA molecule shown in A1) and is related to plant resistance to rice stripe virus. The present invention has experimentally demonstrated that the NP content in rice overexpressing miR-263a is significantly lower than that in wild strains, indicating that overexpressing miR-263a can inhibit the proliferation of RSV virus in plants.

The invention discloses an amylosucrase mutant and application thereof in synthesis of alpha-arbutin, the mutant is obtained by mutation of an amino acid sequence of amylosucrase derived from coprophilobacterium mediterranei, and the amino acid sequence of the mutant is shown as SEQ ID NO.4. The invention further discloses a preparation method of the amylosucrase mutant. Compared with a wild strain, the catalytic activity, the temperature stability and the substrate tolerance of the amylosucrase mutant disclosed by the invention are remarkably improved; the recombinant bacteria constructed by using the recombinant bacteria can be used as a whole-cell catalyst to efficiently catalyze a substrate to synthesize alpha-arbutin in a reaction system containing hydroquinone. Therefore, the amylosucrase mutant disclosed by the invention has a good application prospect.

The invention relates to the technical field of biological medicines, and discloses a construction method and expression application of an Hcp gene deleted strain of a rahnella aqua KCL-5 strain for fish. The Hcp 0186 gene deletion strain is constructed by using a homologous recombination method, the comparison between the deletion strain and a wild strain is carried out, and the result shows that the deletion of the Hcp 0186 gene has an influence on the growth speed, the pathogenicity of the Hcp 0186 gene to the zebra fish is obviously reduced, the survival rate of the infected zebra fish is increased from 0% to 52%, and the Hcp 0186 gene deletion strain can be used as a fish attenuated live vaccine. A recombinant prokaryotic expression vector pET32a-Hcp and expression and purification of fusion protein of the recombinant prokaryotic expression vector pET32a-Hcp are further constructed, and Hcp 0186 rabbit polyclonal antibody serum is prepared as a primary antibody and can be applied to localization expression analysis of fish tissues infected by the bacterium and detection of WB and IHC. Results prove that the Hcp 0186 gene deletion of the strain provides an attenuated strain vaccine and clinical diagnosis and detection application of infected fish of the attenuated strain vaccine, and a scientific basis can be provided for revealing a molecular pathogenic mechanism of the strain and development and application of a mutant strain of the strain.

The invention relates to the technical field of virus detection, in particular to a fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection probe 1106VP and / or 1106WP and primers 1106VWF and 1106VWR. The invention also discloses a fluorescent RT-PCR detection reagent, a fluorescent RT-PCR detection kit, a fluorescent RT-PCR detection probe, a primer, the fluorescent RT-PCR detection reagent and the application of the fluorescent RT-PCR detection kit in identification of a porcine reproductive and respiratory syndrome NJ-1106R strain and a porcine reproductive and respiratory syndrome non-NJ-1106R strain. The probe and the primer disclosed by the invention are high in specificity and high in sensitivity; the accuracy is high, and the accuracy rate of blind sample detection is 100%; the kit is high in universality, can be used for detecting virus liquid for cell culture in a laboratory, and can also be used for detecting porcine serum and tissue samples in clinical samples. The method is an effective method for identifying the PRRSV NJ-1106R vaccine strain and other PRRSV wild strains.

The invention relates to construction and application of an engineering bacterium for high yield of chitinase ChiD, in particular to a bacillus thuringiensis engineering bacterium deltacdsR (P5014-chiD) strain, and the preservation number of the bacillus thuringiensis engineering bacterium deltacdsR (P5014-chiD) strain is GGMCC No.35124. The HD73 endogenous gene chiD gene is efficiently expressed in the bacillus thuringiensis delta cdsR, and the yield in a fermentation culture medium is far higher than that of a wild strain HD73. Compared with HD73 (P5014-chiD), the compound has a relatively strong inhibition effect on mycelial growth of fusarium graminearum and exserohilum turcicum, and shows a better effect on prevention and treatment of larvae of plutella xylostella compared with the HD73 (P5014-chiD).

The application discloses a portable gradient freeze-drying preservation device for wild strain germplasm resources, and relates to the technical field of wild fungus preservation.The application comprises a preservation box, a box cover is rotationally matched with the preservation box, a trapezoidal wild fungus placing rack is slidably matched in the preservation box, and a plurality of universal wheels are arranged at the bottom of the preservation box;an elevating assembly connected with the trapezoidal wild fungus placing rack is arranged on one side in the preservation box, the elevating assembly is matched with the box cover, and an intelligent preservation system is arranged in the preservation box.A temperature gradient control unit is arranged in the application, and a precise controllable temperature gradient can be formed in different level areas of the trapezoidal wild fungus placing rack according to the optimal preservation temperature requirements of different wild fungi.For example, the upper area of the placing rack can be set as a relatively high temperature interval to adapt to some wild fungi which prefer warm environment.

The invention relates to the technical field of virus detection, and particularly discloses a fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection kit for identifying a classical swine fever virus vaccine strain and a wild strain. The kit comprises a specific primer combination, a specific fluorescent probe, a fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) reaction solution, a positive control and a negative control, the primer combination comprises a vaccine strain and wild strain specific primer pair targeting a virus 5 '-UTR region and an E2 gene; the fluorescent probes comprise probes corresponding to the targets respectively and are marked with fluorophores; verification is carried out through double gene targets, missing detection caused by single site mutation is effectively avoided, and the method has the advantages of high specificity, high sensitivity and high accuracy; the invention further provides a non-diagnostic purpose detection method of the kit, detection can be completed within 2 h, and the kit is suitable for rapid identification, epidemic situation monitoring and purification effect evaluation of classical swine fever virus vaccine strains and wild strains in clinical samples.