104 results about "Iridovirus" patented technology

The invention relates to the field of aquaculture, in particular to a preparation method and application of a larimichthys crocea iridovirus recombinant live vector vaccine. According to the invention, replication-deficient type 5 adenovirus is selected as a target gene expression vector, a recombinant adenovirus vector comprising main capsid protein of iridovirus is constructed, recombinant virus is obtained, and the safe and effective large yellow croaker iridovirus disease recombinant live vector vaccine is created. According to the vaccine, the larimichthys crocea is immunized in a soaking or bait feeding mode, the CPM of the larimichthys crocea immunized by the vaccine is 12.0%, and the CPM of the fish without being immunized by the vaccine is 62.0%. In addition, the RPS of the inoculated fish and the RPS of the empty carrier control group fish are 80.65% and 9.68% respectively. The method is at a leading level in the field of pseudosciaena crocea biomedicine at present, and creativity is remarkable.

A spore of a bacillus subtilis B.subtilis WB600 strain is used as an antigen delivery carrier, spore coat protein C (CotC) is used as anchoring protein, a key protein ATPase gene and a CotC gene of micropterus salmoides iridovirus are fused through a gene fusion technology, a recombinant spore surface display system is constructed, the ATPase antigen is stably displayed on the spore surface, and an oral vaccine is prepared. After being orally applied to fishes, the vaccine can tolerate the gastrointestinal tract environment and is accurately delivered to intestinal related lymphatic tissues to stimulate fish immunity, and finally prevention and control of the iridovirus of the micropterus salmoides are achieved.

The invention discloses a multiplex fluorescent quantitative PCR (polymerase chain reaction) detection kit for prawn culture pathogenic bacteria, which is characterized in that the kit contains primers and probes for detecting the prawn culture pathogenic bacteria, wherein the primers and the probes are respectively used for detecting TDH-related hemolsin genes of vibrio parahaemolyticus, ORF107 genes of white spot syndrome viruses, small subunit ribosomal genes of shrimp enterocytozoon hepatopenaei, 37 kDa coat protein genes of infectious subcutaneous and hematopoietic necrosis viruses, DNA (deoxyribonucleic acid) primer genes of full-eye iridovirus 1 and PirA [pi] genes of pathogens of bacterial prawns acute hepatopancreas necrosis; the method has the advantage that the detection of eight prawn culture pathogens can be realized at the same time. The method comprises the following steps of: obtaining a tcdB gene of a high-pathogenicity vibrio and a hemolsin gene of a vibrio harveyi;

The invention discloses a primer probe composition for detecting prawn disease related pathogens, a kit and application of the primer probe composition. The primer probe composition comprises a first primer probe group used for detecting vibrio parahaemolyticus, a second primer probe group used for detecting a tetrapod iridovirus 1, a third primer probe group used for detecting vibrio vitriocultatus and a fourth primer probe group used for detecting enterocytozoon hepatopenaei. The primer probe composition has the characteristics of high sensitivity, good repeatability and strong specificity, and can be used for specifically detecting vibrio parahaemolyticus, tetrapod iridovirus 1, vibrio vitrioculi and enterocytozoon hepatopenaei.

The invention discloses a recombinant yeast for expressing large yellow croaker iridovirus MCP protein and a preparation method and application of the recombinant yeast in preparation of vaccines, and is characterized in that the recombinant yeast is saccharomyces cerevisiae transformed with pYD1-MCP recombinant plasmids and is named as EBY100 / pYD1-MCP, the pYD1-MCP recombinant plasmids can express the MCP protein on a yeast surface display system, the nucleotide sequence of a gene segment of the MCP protein is shown as SEQ ID NO.1, the nucleotide sequence of the gene segment of the MCP protein is shown as SEQ ID NO.2, and the nucleotide sequence of the gene segment of the MCP protein is shown as SEQ ID NO.2. The preparation method comprises a step of constructing a recombinant plasmid pYD1-MCP and a step of constructing a recombinant yeast EBY100 / pYD1-MCP, the oral vaccine of the larimichthys crocea iridovirus vaccine can be prepared, and the oral vaccine has the advantages of being high in protection rate, safe, low in cost, convenient to use and good in application prospect in prevention and control of the larimichthys crocea iridovirus.

The invention discloses a polyclonal antibody of a grouper iridovirus SGIV VP101 gene and application of the polyclonal antibody. The polyclonal antibody is prepared from an SGIVVP101 gene, and the nucleotide sequence of the VP101 gene of an optimized codon is as shown in SEQ ID NO. 1. The polyclonal antibody of the SGIV VP101 gene recombinant expression protein prepared by the invention has good specificity, not only can specifically recognize a prokaryotic expression product of the SGIV VP101, but also can be used for quantitative and positioning analysis of the SGIV VP101 in vivo and in vitro. The successful preparation of the polyclonal antibody of the recombinant expression protein of the SGIV VP101 gene lays a foundation for the establishment of an SGIV immunological detection method, the quantitative and positioning analysis of the protein of the VP101 gene and the analysis of gene functions.

The invention belongs to the technical field of biology, discloses application of naringenin in resisting siniperca chuatsi and frog iridovirus, and discovers the antiviral activity of the naringenin to the siniperca chuatsi and frog iridovirus, the naringenin can down-regulate the expression level of NF-kB and MAPK pathway genes triggered by MRV, effectively inhibit MRV-induced excessive inflammatory response and inhibit MRV proliferation, and the application of the naringenin in resisting the siniperca chuatsi and frog iridovirus. In addition, tissue damage caused by MRV infection can be relieved, and the survival rate of MRV infected fishes is effectively increased; a foundation is provided for preparing a product for resisting the siniperca chuatsi and frog iridovirus by adopting naringenin; meanwhile, the invention also discloses a drug containing naringenin, a fish feed and a fish feed additive.

The invention belongs to the technical field of biology, and discloses application of wogonin in resisting siniperca chuatsi and frog iridovirus, the wogonin can effectively inhibit over-generation of ROS caused by MRV virus infection, and oxidative stress damage of host cells is relieved in a dose-dependent mode. The wogonin can be used for preventing and treating the siniperca chuatsi and frog iridovirus, the high expression state of a proinflammatory gene can be reduced, intestinal tissue damage induced by MRV infection can be relieved, proliferation of MRV in the micropterus salmoides is effectively inhibited, the MRV challenge survival rate of the micropterus salmoides is increased, a basis is provided for preparing a product for resisting the siniperca chuatsi and frog iridovirus from wogonin, and the wogonin has important application value. Meanwhile, the invention also discloses a medicine, a fish feed and a fish feed additive of the wogonin.

The invention relates to the technical field of animal virology, in particular to a target sequence for detecting the virulence of a largemouth bass frog iridovirus and application of the target sequence. The invention provides a target sequence for detecting the virulence of a largemouth bass frog iridovirus. The nucleotide sequence of the target sequence is as shown in SEQ ID NO: 1; the invention also provides a primer pair for detecting the virulence of the largemouth bass frog iridovirus. The detection method provided by the invention can efficiently and accurately distinguish the virulent strain and the virulent strain of the largemouth bass frog iridovirus, the detection time can be completed within 1.5 hours, a powerful support is provided for accurate judgment of the virulence of the largemouth bass frog iridovirus, and an effective means is also provided for screening the virulent strain and the virulent strain of the largemouth bass frog iridovirus.

The invention discloses a targeted delivery system of a largemouth bass frog iridovirus oral vaccine and a preparation method of the targeted delivery system, and relates to the technical field of biological medicines.The method comprises the following steps that a starch solution and a largemouth bass frog iridovirus inactivated vaccine are stirred and mixed, then a paraffin emulsion is added, stirring and emulsifying are conducted, and a W / O emulsion is obtained; adding calcium chloride, stirring and mixing, centrifuging, and resuspending by using a PBS (Phosphate Buffer Solution) to obtain a microsphere resuspension solution; stirring and mixing with a quaternized chitosan solution, adding glutaraldehyde for crosslinking, centrifuging, and freeze-drying. The invention further discloses a targeted delivery system of the largemouth bass frog iridovirus oral vaccine prepared by the method. The vaccine is orally administered after being microencapsulated, so that the human cost is low, the stimulation to fish bodies is small, the problems of inconvenience in fish administration, low bioavailability of oral administration, instability in gastrointestinal tracts and the like are solved, and the vaccine has an excellent application prospect in the field of LMBV prevention and control.

The invention discloses an application of bortezomib in preparation of a medicine for resisting grouper iridovirus. Researches show that bortezomib has a remarkable inhibiting effect on grouper iridovirus, and is low in cytotoxicity and good in safety. The bortezomib can obviously reduce the fluorescence signal intensity of virus protein, inhibit the transcriptional level of main capsid protein MCP and envelope protein VP19 of the virus and reduce the copy number of DNA and mRNA of the virus; the bortezomib can effectively inhibit grouper iridovirus infection and effectively block synthesis of virus protein, so that efficient inhibition of grouper iridovirus infection is achieved, and along with prolonging of virus infection time, bortezomib can also remarkably inhibit virus gene transcription, protein synthesis and genome replication. Therefore, bortezomib not only can effectively prevent and treat grouper iridovirus infection, but also has the characteristics of high specificity, low toxicity, remarkable antiviral effect and the like, and also has important application value in prevention and control of iridovirus-related diseases in aquaculture industry.

The invention discloses an application of Micropterus salmoides Piscidin polypeptide in preparation of an anti-fish virus drug. The amino acid sequence of the polypeptide is as shown in SEQ ID NO. 2 or SEQ ID NO. 3. The research finds that the micropterus salmoides Piscidin is an antibacterial peptide with an antiviral function, the polypeptide synthesized by the artificially synthesized micropterus salmoides Piscidin has the characteristic of inhibiting fish DNA virus replication, has a remarkable inhibiting effect on fish iridovirus replication at low concentration, and can be applied to preparation, research and development of fish antiviral drugs. Research of the antibacterial peptide has important scientific value and application value for further research and development of disease-resistant functional gene products.

The invention relates to a rapid detection method for nucleic acid of iridovirus of Micropterus salmoides and a kit thereof, belonging to the technical field of molecular biology, the rapid detection method adopts a two-step method of RPA-Cas12a for treatment, can be used for fluorescence detection through fluorescence labeling and also can be used for detection through an immune colloidal gold test strip, and is simple and convenient to operate, rapid and sensitive, strong in specificity, good in stability and low in cost.

The invention provides micropterus salmoides Hepcidin-1 (Ms-Hepcidin-1) and an application of a synthetic polypeptide of the micropterus salmoides Hepcidin-1 in preparation of an anti-fish virus drug. The research finds that the Ms-Hepcidin-1 synthetic polypeptide has a remarkable antiviral effect and is a novel antibacterial peptide with a function of inhibiting fish viruses. The Ms-Hepcidin-1 synthetic polypeptide and the recombinant eukaryotic expression vector of the Ms-Hepcidin-1 synthetic polypeptide can obviously reduce the transcription level of MCP, MMP and DNMT genes and the MCP protein level of LMBV, inhibit the replication of the LMBV and reduce the virus titer, and have an obvious fish virus resisting effect. Moreover, the polypeptide synthesized by Ms-Hepcidin-1 is simple to prepare, low in cost, safe to use and convenient to preserve, can be used as a novel antiviral functional gene product to prepare antiviral drugs for fishes, and has important significance on prevention and control of fish iridovirus diseases.

The application discloses an anti-LMBV single-chain antibody and application thereof in preparation of a quantum dot fluorescent immunochromatography test strip, and belongs to the technical field of biological detection. The single-chain antibody comprises Ab-K1 and Ab-K2, the amino acid sequence of the Ab-K1 is shown as SEQ ID NO. 11, and the amino acid sequence of the Ab-K2 is shown as SEQ ID NO. 12. The single-chain antibody used in the application can accurately recognize and combine with iridovirus in a sample to be detected, so that the accuracy of detection is ensured. Meanwhile, the antibody labeled with fluorescent quantum dots has good fluorescent characteristics, so that the fluorescent signal is easy to observe and detect, and the specific surface area is relatively large, so that the antibody can be combined with more virus molecules, so that the sensitivity of detection is improved, the accuracy and sensitivity of detection are improved, the operation steps are simplified, and powerful technical support is provided for detection and prevention and control of iridovirus.

The invention relates to a RAA-CRISPRCas13a method for detecting iridovirus fluorescence of micropterus salmoides, and relates to the technical field of biological detection.The method comprises the following steps that an RAA primer pair and guide crRNA are designed and synthesized based on a virus specific gene sequence; the method comprises the following steps: constructing a single-tube reaction system containing an RAA amplification component, Cas13a protein, crRNA and a fluorescent reporter molecule; after sample nucleic acid is added, constant-temperature incubation is carried out for 15-30 minutes at 37-42 DEG C, an RAA amplification product activates trans-cleavage activity of Cas13a, and a cleavage reporter molecule generates a fluorescence signal; and virus judgment is realized by detecting a fluorescence signal. According to the method, isothermal amplification and CRISPR detection are efficiently integrated through a single-tube reaction system, and the method has the advantages of being easy and convenient to operate, rapid in detection, high in sensitivity and high in specificity and is suitable for on-site high-throughput screening.

The invention belongs to the technical field of aquatic pathogen detection, and provides a primer and probe combination with strong specificity and high sensitivity, a kit and a cdPCR detection method in order to solve the problems of dependence of absolute quantification of an existing qPCR technology on a standard curve and low repeatability and stability caused by uncontrollable quality of a standard substance. The primer probe can generate specific amplification on LYCIV and has no cross reaction on other common aquatic pathogens (such as NNV, DIV1 and the like), so that the specificity of a detection result is fundamentally ensured, a false positive result caused by the cross reaction is effectively avoided, and the diagnosis accuracy is improved. The method has the characteristics of extremely high sensitivity and absolute quantification, the lower limit of detection reaches 6.2 copies / mu L, the method has more advantages in quantification of samples with extremely low concentration, early diagnosis and detection of extremely low virus load can be realized, viruses with extremely low content in fish bodies can be detected earlier, precious time is provided for disease early warning and early intervention, and the method is worthy of popularization and application. Disease outbreak is effectively prevented.

The invention provides an application of an Opnlrp12 gene in prevention and treatment of torus schlegeli virus and / or bacterial infection, and belongs to the technical field of gene engineering. Through targeted knockout of Opnlrp12, it is found that the expression level of a disease-resistant gene is also reduced, and therefore it is proved that Opnlrp12 can effectively inhibit infection caused by viruses and / or bacteria, and the survival rate of oplegnathus punctatus is increased. By means of the method, control over inflammatory diseases caused by iridovirus and / or vibrio harveyi can be achieved, and a target spot is provided for treatment of iridovirus infection of oplegnathus punctatus.

The application provides a preparation method of a shrimp iridovirus DIV1 neutralizing antibody and application thereof. The gene sequence shown as SEQ ID NO. 1 and the amino acid sequence shown as SEQ ID NO. 2 are used to solve the problem that there is no neutralizing antibody for the shrimp iridovirus DIV1 at present. The antibody for the shrimp iridovirus DIV1 prepared by the application can specifically recognize the DIVI virus and has good virus neutralizing activity, can effectively reduce the death of shrimps caused by DIV1 virus infection, significantly improve the ability of shrimps to resist DIV1 virus infection, and effectively prevent the infection and transmission of the virus.

The invention discloses an application of alpha-lipoic acid in preparation of an anti-iridovirus drug. Related experiments show that alpha-lipoic acid can safely and effectively treat iridovirus such as frog virus type 3, not only can effectively reduce harm of iridovirus to frog and fish culture, but also can promote ecological high-quality development of aquaculture industry, and has obvious economic and social benefits.

The invention provides a method for identifying the iridovirus damage grade of micropterus salmoides based on machine vision assistance, and belongs to the technical field of aquatic product infection identification. According to the method, the damage degree of micropterus salmoides infected with iridovirus is measured in a standardized mode, early-stage micropterus salmoides body surface damage data are analyzed based on visual machine assistance, body surface damage areas are divided, the body surface damage number and damage area are measured, individual comprehensive damage scores are calculated through data standardization and PCA dimension reduction, and body surface damage grades are constructed. And a scientific basis is provided for disease management of the micropterus salmoides.

This invention provides the application of Hepcidin-1 (Ms-Hepcidin-1) and its synthetic polypeptide in the preparation of antiviral drugs for largemouth bass. The invention reveals that the Ms-Hepcidin-1 synthetic polypeptide exhibits significant antiviral activity and is a novel antimicrobial peptide with the function of inhibiting fish viruses. Both the Ms-Hepcidin-1 synthetic polypeptide and its recombinant eukaryotic expression vector can significantly reduce the transcriptional levels of MCP, MMP, and DNMT genes and the MCP protein level of LMBV, inhibit LMBV replication, and reduce viral titers, demonstrating significant antiviral activity against fish viruses. Furthermore, the polypeptide synthesized from Ms-Hepcidin-1 is simple to prepare, low in cost, safe to use, and easy to store. As a novel antiviral functional gene product, it can be applied to the preparation of antiviral drugs for fish, which is of great significance for the prevention and control of iridovirus disease in fish.

The invention provides application of epicatechin in preparation of a medicine for resisting frog iridovirus. Related experiments prove that the epicatechin can efficiently prevent and control infection and transmission of the frog iridovirus, so that the harm of the frog iridovirus to healthy culture of frogs such as bullfrogs can be reduced to a great extent, and high-quality ecological culture and economic benefits of the frog culture industry are promoted.

The invention relates to the technical field of gene detection, in particular to a rapid shrimp iridovirus detection primer group based on an RPA-CRISPR / Cas12a system, application and a kit. The primer group comprises an iridovirus constant-temperature gene amplification primer and a specific recognition amplification product target sequence probe; wherein the iridovirus constant-temperature gene amplification primer, the specific recognition amplification product target sequence CRISPR probe and the ssDNA reporter probe are recorded in the specification. An isothermal amplification primer and CrRNA are designed based on the shrimp red color virus gene, the target gene is amplified through the isothermal amplification technology, Cas12a is guided by the CrRNA to accurately recognize and activate trans-cleavage, a test strip report probe is synchronously split, and the result is interpreted by naked eyes within 30 min by virtue of the visual advantage of an immune test strip. The whole process does not need large instruments, the operation is simplified, the cost is low, and a visual and efficient solution is provided for on-site rapid diagnosis.

The present invention provides a nano-antibody targeting giant salamander iridescent virus and its preparation method and application; the invention aims to provide a nano-antibody targeting giant salamander iridescent virus with low expression cost, low difficulty, stable source, and the ability to effectively bind to giant salamander iridescent virus and specifically realize immune analysis of giant salamander iridescent virus; the amino acid sequence of the nano-antibody is shown in SEQ ID NO.1-6 and belongs to the field of biological science and technology.

The invention relates to the field of material science and engineering, and particularly discloses a screening method of an anti-macrobrachium rosenbergii iridovirus natural compound based on an MCP protein target. The method comprises the following steps: constructing a three-dimensional structure of a virus major capsid protein MCP and determining an active pocket; performing structure pretreatment on the natural compound library; by taking the active pocket as a target spot, performing multiple rounds of virtual screening through molecular docking to obtain a candidate compound with binding energy less than or equal to-10.0 kcal / mol; and establishing a macrobrachium rosenbergii artificial infection model, determining the virus inhibition rate and protection rate of the candidate compound, and screening the active compound. The eugenol, the amentoflavone, the polyphyllin II and the solamargine screened by the method show a remarkable antiviral effect in an in-vivo injection test and breeding verification. According to the method, accurate screening from target spot prediction to in-vivo verification is realized.

The invention provides a primer probe group, a kit and a detection method for detecting microsporidia of eriocheir sinensis, and belongs to the technical field of pathogen detection. The invention provides a primer probe group for detecting microsporidia of eriocheir sinensis. The primer probe group comprises a primer pair and a TaqMan probe, the nucleotide sequences of the primer pair and the TaqMan probe are as shown in SEQ ID NO: 1 to SEQ ID NO: 3. In the invention, the primer probe group can specifically detect the eriocheir sinensis microsporidia, and does not have cross reaction with aeromonas veronii, vibrio parahaemolyticus, eriocheir sinensis spiroplasma, white spot syndrome virus and siniperca chuatsi iridovirus. The lowest detection limit of the primer probe group is 4.6 * 10 < 1 > copies / mu L, the sensitivity is improved by 1000 times compared with that of common PCR, and great detection advantages are shown.

The present invention relates to a multiplex PCR primer set for simultaneously detecting four pathogens, as well as a detection method and application thereof, belonging to the field of molecular biology. The four pathogens are neural necrosis virus, iridovirus, Streptococcus iniae, and Vibrio harveyi. The primer set comprises SEQ ID NOs. 1-8. Using the primers of the present invention, neural necrosis virus, iridovirus, Streptococcus iniae, and Vibrio harveyi can be simultaneously detected in a short period of time, thereby achieving the purpose of establishing a simple and effective method for simultaneously detecting multiple fish pathogens. Furthermore, the detection primer set and detection method of the present method have good specificity and high sensitivity.

This invention belongs to the field of new drugs, specifically relating to a pyran derivative, B63, and its use in the preparation of drugs for use against largemouth bass frog iridescence virus. The invention provides the use of B63, its derivatives, or pharmaceutically acceptable salts thereof, or substances containing B63, its derivatives, or pharmaceutically acceptable salts as active ingredients, in the preparation of products for use against largemouth bass frog iridescence virus. B63 exhibits both in vitro and in vivo anti-largemouth bass frog iridescence virus activity and is a potential drug candidate for use against largemouth bass frog iridescence virus.

The present application relates to a new application of radix cirsii japonici extract in resisting largemouth bass iridovirus. The present application finds that the radix cirsii japonici extract has a significant effect in resisting largemouth bass iridovirus, can significantly reduce the copy number of largemouth bass iridovirus, inhibit the expression of largemouth bass iridovirus MCP protein, improve the phagocytic ability of largemouth bass kidney leukocytes, and effectively improve the survival rate of fish infected with largemouth bass iridovirus, and has good biological safety. Therefore, the radix cirsii japonici extract can be used for preparing products such as drugs, fish feed or fish feed additives resisting largemouth bass iridovirus, and provides an effective prevention and control means for largemouth bass and other fish breeding.