33 results about "Koi herpesvirus" patented technology

The invention discloses an LAMP detection kit for Koi herpesvirus and a detection method. The LAMP detection kit comprises a 10*ThermoPol Reaction buffer liquor, Bst DNA (Deoxyribose Nucleic Acid) polymerase, dNTPs, outer primers F3 and B3, inner primers FIP and BIP, Betaine, MgCL2, and 1000*SYBR Green I. The LAMP detection kit provided by the invention has the characteristics of simplicity, convenience, rapidness and high specificity and sensitivity, and KHV (Koi Herpesvirus) in a sample can be accurately detected within 2 hours by a water bath kettle or a metal bath. The diseased fish tissue infected by KHV can be detected, and cells infected by KHV can be detected, so that the kit is very suitable for field fast detection of KHV.

The invention discloses a koi herpesvirus RT-LAMP (reverse transcription loop-mediated isothermal amplification) detection primer group, a kit and a detection method thereof. The detection primer group comprises a pair of outer primers, a pair of loop primers and a pair of inner primers; the detection kit comprises a primer group, an RT-LAMP reaction liquid, Bst DNA (deoxyribonucleic acid) polymerase and negative/positive control. The detection method of the detection kit comprises the following steps: extracting DNA of a to-be-detected virus, amplifying the sample DNA template at 63-65 DEG C with six specific primers and the Bst DNA polymerase with strand displacement activity, observing whether an S amplification curve exists through a convenient constant temperature fluorescence detector, and determining whether the to-be-detected sample contains koi herpesvirus (KHV) DNA. The kit has the advantages of high specificity, high sensitivity, rapidity and high efficiency, simplicity and convenience in operation, easily readable result, and so on, and is suitable for being generalized and applied to detection units of different levels and aquaculture users.

The invention discloses a koi herpesvirus ORF25 nucleic acid vaccine, which has good immunogenicity. According to the koi herpesvirus ORF25 nucleic acid vaccine, ORF25 gene is cloned to an eukaryotic expression vector pIRES-neo to obtain recombined plasmid. The koi herpesvirus ORF25 nucleic acid vaccine has effective prevention effect to koi herpes.

The invention discloses a KHV (koi herpes virus) CPA (cross priming amplification) detection primer and an application. A KHV CPA detection kit comprises following ingredients: 10*ThermoPol Reaction Buffer, Bst DNA polymerase, dNTPs, cross primers CPF and CPR, detection primers DF and DR, stripping primers DPF and DPR, MgSO4, Betaine and a nucleic acid test strip. The KHV CPA detection primer has the characteristics of simplicity, convenience, rapidness and high specificity and sensitivity and is objective and direct in determination of results, low in cost, convenient to use and very safe to people and the environment. The KHV CPA detection primer not only can be used in a special laboratory, but also can be applied to rapid onsite detection in the field, only one metal bath or water bath kettle is required, and a KHV in a sample can be accurately detected within 1.5 h.

The invention discloses a preparation method and application of a monoclonal antibody against koi herpesvirus (KHV). The prepared monoclonal antibody against KHV is secreted by a mouse hybridoma cellstrain of which the preservation number is CCTCC NO: C201887, and the titer of the monoclonal antibody is 1:(1.28 is multiplied by 105); and the hybridoma cell strain has high activity, can recognizethe structural protein of the KHV, and has strong specificity. On the basis, a sandwich ELISA detection method for detecting the KHV is established, and the minimum detection concentration is 3.923ng/mL. The monoclonal antibody is utilized for establishing an indirect sandwich ELISA kit for detecting the KHV, and the kit can be used for large-scale detection of the KHV and has wide application prospects.

The invention discloses a CNT (carbon nano tube)-loaded nucleic acid vaccine based on a KHV (Koi herpesvirus) ORF149 and an application. A single-walled CNT-loaded pcDNA-ORF149 nucleic acid vaccine system is constructed successfully, fish antibody level is improved, the relative percent survival of the system is 81.9%, the immune level is far higher than that of other immune systems, and the system has the application potential in aquaculture and pathogen research.

The invention discloses primers and method for fast detecting koi herpesviruses (KHV) and belongs to the field of animal pathogen detection. The primers and method are high in specificity and sensitivity and good in repeatability. According to the technical scheme, the nucleotide sequence of the primers is as shown in SEQ ID No. 1 and SEQ ID No. 2, and the nucleotide sequence of a probe is as shown in SEQ ID No. 3. The detecting method includes the steps of firstly, extracting virus genome DNA from a clinical sample; secondly, using the extracted sample DNA as the template, using the primer group in claim 1 and the probe in claim 2 to prepare an RPA reaction system, and using a TwistAmp exo kit to perform RPA amplification reaction; thirdly, placing the reaction tube after the RPA amplification into a fluorescence detector, monitoring fluorescent signals in real time, determining that the sample contains KHV if an evident fluorescent signal curve exists, and determining that the sampledoes not contain KHV if no fluorescent signal curves exist.

The invention discloses a dual real-time fluorescence quantitative PCR (polymerase chain reaction) detection method for a CEV (carp edema virus) and a KHV (Koi herpesvirus). The method comprises following steps: step one, synthesis of primers and a TaqMan probe: a primer sequence is optimized according to a CEV P4a gene sequence, first base of an upstream primer is changed into degenerate base W,and first base of a downstream primer is changed into degenerate base R; a pair of specific primers and one TaqMan probe are designed according to KHV ORF7 gene conserved sequence; FAM and VIC are taken as probe reporter groups and BHQ1 is taken as a probe quenching group; step two, determination of a reaction system and conditions: a 20 mu L reaction system is adopted and determined as 10 mu L 2*Probe PCR Master Mix, final concentration of the upstream and downstream primers and the probe primers is in a range of 0.2 mu mol/L-0.8 mu mol/L, a QN ROX reference dye is 0.1 mu L, a template is 2.5mu L and the balance of DEPC water, and the total amount is 20 mu L; the reaction procedure is that predegeneration at 95 DEG C is performed for 2 min, and one cycle is executed; annealing is performed for 5 s at 95 DEG C and performed for 31 s at 50-60 DEG C, and 40 cycles are executed.

The present invention provides a DNA vaccine for carps for inducing protective immunity against Koi herpesvirus (KHV). The DNA vaccine comprises a DNA comprising a nucleotide sequence encoding an immunogenic polypeptide against Koi herpesvirus (KHV) of carps, or an expression vector comprising the DNA as an active ingredient.

The invention relates to the technical field of aquatic antiviral micro-ecological preparations, and especially relates to a bacillus velezensis YFI-4, and application thereof in preparation of drugsfor treating viral diseases of freshwater cultured animals. The preservation number of the bacillus velezensis disclosed by the invention is CCTCC NO: M2019653. The bacillus velezensis YFI-4 disclosedby the invention is used for fermenting supernatant, reduces infection activity of koi herpesvirus (KHV), cyprinid herpesvirus type 2 (CyHV-2), giant salamander iridovirus (GSIV) and the like on hostcells, inhibits infection of virus invasion on the cells is inhibited, and has no toxic or side effect on the cells and can be used for preventing and treating viral diseases of freshwater cultured animals.

The invention discloses a koi herpesvirus RPA primer pair, a probe and a detection kit and application thereof. Sequences of the RPA primer pair are as shown in SEQ ID NO.4-19, and the sequences of the probe are as shown in SEQ ID NO.2-3. The probe and the primers provided by the invention are high in sensitivity, stable in detection effect and high in specificity. On the basis, the constructed detection reagents and detection kit have very high detection sensitivity, the lowest detection limit is 100 copy/mu L, and the detection time is only 4-18 minutes. Compared with KHV detection gold standard PCR and real-time PCR, the detection reagents and the detection kit have the advantages of time and detection accuracy, with extremely high application value.

The invention relates to the technical field of aquatic product antiviral micro-ecological preparations, and in particular relates to bacillus licheniformis YFI-2 and an application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals. The preservation number of the bacillus licheniformis disclosed by the invention is CCTCC NO: M2019652. A fermenting supernatant of the bacillus licheniformis YFI-2 disclosed by the invention can reduce infection activity of grass carp reovirus (GCRV), koi herpesvirus (KHV), cyprinid herpesvirus type 2 (CyHV-2), ictalurus punctatus reovirus (CCRV), giant salamander iridovirus (GSIV) and the like on host cells, can inhibit infection on cells by virus invasion, has no toxic or side effect on cells, and can be used for preventing and treating viral diseases of freshwater cultured animals.

The invention discloses a visual rapid detection kit for KHV (Koi herpesvirus). The visual rapid detection kit comprises a reagent A, a reagent B, a reagent C, a reagent D, a reagent E, a reagent F, areagent G and a detection tube containing a reagent H, wherein the reagent A is a tissue lysate I; the reagent B is a tissue lysate II; the reagent C is isopropyl alcohol; the reagent D is aqueous ethanol with volume percentage concentration being 65%-80%; the reagent E is sterilized double distilled water; the reagent F is 90-110 mg of nucleic acid extracting magnetic beads resuspended in 1 ml of double distilled water; the reagent G is KHV positive plasmid. The operation process of the kit is simple, KHV detection can be completed in 1.5-2 h, and the KHV detection can be completed only by one water bath kettle or metal bath without any high-end instruments and equipment.

The invention discloses an RT-PCR (reverse transcription polymerase chain reaction) detection kit and method of koi herpesvirus. A primer group includes primers S1 and S2, wherein S1 is 5'-AAGGCTTAGTAGCTTACCAT-3', S2 is 5'-CAAGGTAGCTGAGTCGACCTATG-3', and S3 is 5'-ATAAGGGCTGAGTCCTGAT-3'. The specific primers S1, S2 and S3 are designed and capable of carrying out specific amplification on koi herpesvirus. The detection kit as a special kit has high sensitivity, reaches 100% in coincidence rate as compared to virus separation, IFA (indirect immunofluorescent assay) and other methods, and is widely applicable to identification diagnosis of koi herpesvirus.

The invention relates to a Chinese herb composition capable of effectively treating Koi herpesvirus, wherein the Chinese herb composition comprises, by weight, 55-65 parts of eucalyptus leaf, 35-45 parts of scutellaria baicalensis, 45-55 parts of radix isatidis, 45-55 parts of honeysuckle, 20-35 parts of sophora flavescens ait, 15-25 parts of cassia twig, 5-15 parts of cimicifuga foetida, 2-15 parts of nepeta cataria l, 10-25 parts of peppermint, 5-15 parts of agastache rugosa, and 10-25 parts of folium isatidis. According to the present invention, the components are subjected to scientific compatibility according to the traditional Chinese medicine principle of assistant and guide, the effective components of most of the herbs are subjected to quantitative and semi-quantitative detectionby using the modern scientific and technological means, and the prepared finished product can provide good treatment effect for Koi herpesvirus; various Chinese herbs acts synergistically, such that the inhibition effect on viruses is enhanced, and the toxicity of the single herb is reduced; and the Chinese herb composition can significantly prevent and treat Koi herpesvirus, is the traditional Chinese medicine with characteristics of good treatment effect, no pollution, no residue and easy use, and is especially suitable for the large-scale industrial breeding of Cyprinus carpio.