28 results about "Hepatitis E virus" patented technology

The invention relates to the technical field of molecular biology, and discloses a hepatitis E virus rapid detection method and system based on an animal detection technology, and the method comprises the following steps: carrying out nucleic acid extraction on a hepatitis E virus sample to obtain a nucleic acid extraction sample; establishing a PCR system of the nucleic acid extraction sample, and generating uniform microdroplets of the nucleic acid extraction sample and the PCR system; calculating a rupture coefficient of the uniform microdroplet, performing PCR amplification on the uniform microdroplet to obtain an amplified microdroplet, and performing signal detection on the amplified microdroplet by using a preset fluorescent dual-channel to obtain a fluorescent detection signal; generating a two-dimensional scatter diagram of the amplified droplets to determine FAM positive droplets in the amplified droplets; and analyzing the HEV genotype and variation site of the hepatitis E virus sample to establish an HEV strain evolutionary tree and risk-inducing factors of the hepatitis E virus sample, and generating an HEV geographical distribution heat map of the to-be-detected region by combining the HEV strain evolutionary tree and the risk-inducing factors. According to the invention, the accuracy of rapid detection of hepatitis E virus can be improved.

The invention discloses a digital PCR nucleic acid detection kit, and relates to the technical field of in vitro diagnosis detection, the kit comprises primers, probes, a reaction liquid and a quality control liquid, the kit is used for simultaneously detecting hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus and hepatitis E virus, the reaction liquid comprises Master Mix and sterile enzyme-free water, and the quality control liquid is used for simultaneously detecting the hepatitis A virus, the hepatitis B virus, the hepatitis C virus, the hepatitis D virus and the hepatitis E virus. The quality control liquid comprises a negative quality control product and a positive quality control product. Through cooperation of sample collection, sample treatment, digital PCR reaction system preparation, digital PCR amplification and result analysis, nucleic acid detection based on the PCR technology shortens the window period by 50-70% through direct detection of virus genetic materials, becomes a key means for early diagnosis, and adopts the technical principle of digital PCR to realize rapid detection of the virus genetic materials. Compared with a traditional fluorescent quantitative PCR and immunological detection method and a qPCR method, the detection sensitivity is improved by 10-100 times, and absolute quantification can be achieved.

Described herein are compositions, uses thereof, and methods for treating a viral infection in a host cell or organism infected by the virus, such as coronaviruses (e.g., severe acute respiratory syndrome coronavirus [SARS-CoV], severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2, the virus and its mutant forms that cause COVID-19], Middle East respiratory syndrome coronavirus [MERS-CoV]), Zika virus, Lassa virus, Crimean Congo hemorrhagic fever virus, hepatitis E virus, and other RNA viruses. Also described herein are synthetic rocaglate compositions, uses thereof, and methods for reducing or inhibiting translation initiation of a messenger ribonucleic acid (mRNA) of a virus in a host cell or organism infected by the virus.

Described herein are compositions, uses thereof, and methods for treating a viral infection in a host cell or organism infected by the virus, such as coronaviruses, Zika virus, Lassa virus, Crimean Congo hemorrhagic fever virus, hepatitis E virus, and other RNA viruses. Also described herein are synthetic rocaglate compositions, uses thereof, and methods for reducing or inhibiting translation initiation of a messenger ribonucleic acid (mRNA) of a virus in a host cell or organism infected by the virus.

The invention relates to the technical field of DNA vaccines, in particular to a hepatitis E virus targeting antigen binding fragment and application thereof in preparation of a DNA vaccine. The base sequence of the antigen binding fragment is shown as SEQ ID NO. 1, and the amino acid sequence of the antigen binding fragment is shown as SEQ ID NO. 2; and the molecular structure is 6 * His labelle-ORF2 antigen fragment-6 * His label. According to the scheme, the DNA vaccine with stable quality can be rapidly prepared, and cellular immunity and humoral immunity protection can be provided for an organism.

The application provides a hepatitis E virus nucleic acid detection primer, probe and kit, wherein according to a target sequence of a hepatitis E virus gene, a primer and a probe sequence in an asymmetric primer ERA detection system are designed, screened, and optimized in terms of an upstream primer ratio, a downstream primer ratio and an amplification reaction system, so that the primer and the probe can specifically recognize a nucleic acid sequence of the hepatitis E virus, non-specific binding with other irrelevant nucleic acids is reduced, and thus the specificity of detection is improved; meanwhile, efficient amplification of the hepatitis E virus nucleic acid is realized, and even in the case of extremely low content of the virus nucleic acid, the hepatitis E virus nucleic acid can be accurately detected, and thus the sensitivity and the amplification efficiency of detection are improved.

The invention belongs to the technical field of biological detection, and particularly relates to a primer probe combination, a kit and a method for detecting eight porcine viruses. Specific primers and probes are designed for porcine circovirus type 2, porcine circovirus type 3, African swine fever virus, rotavirus, porcine hepatitis E virus, classical swine fever virus, norovirus and fiveleaf virus, and the kit is used for detecting eight viruses of pigs and has the advantages of high specificity, high sensitivity, stable effect and simplicity and convenience in operation; according to the method, eight infection pathogens can be detected, and the method is of great significance to the domestic pig feeding industry and wild boar epidemic disease monitoring and controlling work.

The application provides mRNA based on HEV ORF2 truncated protein, a preparation method and application thereof, the mRNA based on HEV ORF2 truncated protein includes 5'UTR region, SP fragment, 3'UTR region, 3' terminal poly A tail, gene for encoding hepatitis E virus ORF2 truncated protein and 5' cap structure, the hepatitis E virus ORF2 truncated protein is ORF2-239aa protein or ORF2-549aa protein, the nucleotide sequence of the gene for encoding the ORF2-239aa protein is shown as SEQ ID NO.1, the nucleotide sequence of the gene for encoding the ORF2-549aa protein is shown as SEQ ID NO.2, the application has application value in hepatitis E research and vaccine creation, and can greatly promote the transformation application of vaccine.

The invention discloses a new application of lenalidomide (CC-5013) in the treatment of male oligospermia and azoospermia caused by hepatitis E virus (HEV) infection or the improvement of sperm quality, and particularly discloses a new application of lenalidomide (CC-5013) in the treatment of male oligospermia and azoospermia caused by hepatitis E virus (HEV) infection. The lenalidomide is proved to have an obvious treatment effect on sperm quality reduction caused by HEV infection through sperm staining, sperm motility detection, testis pathology analysis, quantitative detection of sperm development key genes and other methods by utilizing an HEV infected mouse model. The treatment scheme provided by the invention is safe and effective, good in treatment effect and low in medicine cost, is suitable for rapid treatment of male reproductive injuries such as male azoospermia, oligospermia and poor sperm quality caused by virus infection, and provides medicine support for diagnosis and treatment of male infertility patients caused by HEV infection.

Virus-Like Particles derived from the subfamilies, Parahepevirinae, which infect trout and salmon, and the Orthohepevirinae, which infect mammals and birds, particularly those of the species Paslahepevirus balayani, which can cause acute hepatitis in humans and several mammalian species, and chronic conditions in immunocompromised patients are also disclosed. Major aspects of the invention relate to compositions of Virus-Like Particles comprising viral capsid proteins capable of assembly in cultured cells that may be purified, disassembled, and reassembled in the presence of other molecules suitable for use as therapeutic drug products to facilitate the targeting and delivery of cargo molecules to specific cells or tissues, or as antigenic agents designed to stimulate responses to heterologous epitopes exposed on the surfaces of Virus-Like Particles. Preferred aspects relate to functional capsids comprising polypeptide sequences comprising one or more amino acid substitutions, insertions, or deletions of amino acid encoded by a consensus of ORF2 genes, wherein said variant polypeptides are functionally-similar or have enhanced properties compared to capsid polypeptides encoded by naturally-occurring viruses obtained from clinical samples or prototype Hepatitis E Viruses (HEV). Other aspects include the design and assembly of modified vectors to facilitate the basic and applied studies leading to the development and commercialization of novel drug products, and as tools advancing the interests of institutions involved in animal and human healthcare.

The invention provides a bivalent vaccine which comprises a rabbit hemorrhagic syndrome virus antigen, a rabbit hepatitis E virus antigen and a microbial freeze-dried slow-release agent, and provides a preparation method which comprises the following steps: culturing the rabbit hepatitis E virus by using passage cells, obtaining the rabbit hepatitis E virus antigen, inoculating the rabbit hemorrhagic syndrome virus, and carrying out freeze-drying on the rabbit hemorrhagic syndrome virus. A rabbit hemorrhagic syndrome virus antigen is obtained, the rabbit hemorrhagic syndrome virus antigen and a rabbit hepatitis E virus antigen are mixed in proportion, a freeze-drying protective agent is supplemented, and the bivalent vaccine is obtained. The invention further provides an application experiment result of the compound, and the compound is used for preventing or treating rabbit hemorrhagic syndrome and rabbit hepatitis E. According to the invention, the two antigens are proportioned properly, so that the prepared vaccine at least achieves the efficacy of preventing and controlling immune epidemic diseases. The bivalent vaccine provided by the invention can prevent two diseases by one injection, reduce immune stimulation, continuously enhance the immune effect of a body, promote the body to generate better immune response, reduce the immune cost, simplify the immune procedure, and is more economical and reliable.

This application relates to a method for preparing and using a hepatitis E virus DNA vaccine. The vaccine comprises an antigen-binding fragment, the base sequence of which is shown in SEQ ID NO. 1. The solution provided herein enables rapid and cost-effective preparation of a high-quality, stable vaccine, providing rapid and efficient cellular and humoral immune protection.

The invention discloses a primer group, a kit and a detection method for detecting a meriones unguiculatus hepatitis E virus, and belongs to the technical field of virus detection. And the primer group comprises a primer Cl-HEV-F, a primer Cl-HEV-R, a primer Cl-HEV-qF, a primer Cl-HEV-qR and a primer Cl-HEV-qP. The detection method comprises the following steps: extracting virus RNA and reversely transcribing into cDNA; the method comprises the following steps: amplifying a capsid protein gene by using a Cl-HEV-F / R primer PCR (Polymerase Chain Reaction), connecting with a pMD19-T vector, and transforming DH5alpha cells for screening. The optimal conditions of RT-qPCR are as follows: the primer / probe concentration is 0.4 mu mol / L, and the annealing temperature is 60 DEG C. The method is high in specificity, the sensitivity reaches 4.33 * 10 copy number / mu L, and the repeatability is good. The detection system has high sensitivity, high specificity and good repeatability, and can be used for detection and quantitative analysis of the meriones unguiculatus hepatitis E virus.

The invention relates to a primer group, a probe and a detection method for a lesser panda-derived hepatitis E virus and a parainfluenza 5 virus, the primer group and the probe can simultaneously and specifically recognize the lesser panda-derived hepatitis E virus and the parainfluenza 5 virus by a one-tube method, do not generate cross reaction with other common pathogens, have high specificity and sensitivity, and can be used for detecting the lesser panda-derived hepatitis E virus and the parainfluenza 5 virus. According to the detection method formed on the basis of the primer group and the probe, detection can be completed within 20 min under the constant-temperature condition, and a rapid pathogen diagnosis method and apparatus are provided for zoos, wild animal rescue centers and other conservation institutions.

The invention relates to a preparation method of a specific antigen binding fragment and application of the specific antigen binding fragment in a hepatitis E virus DNA vaccine. The base sequence of the antigen binding fragment is as shown in SEQ ID NO. 1. According to the antigen binding fragment and the scheme for preparing the vaccine, the vaccine with stable quality can be rapidly prepared, and cellular immunity and humoral immunity protection can be provided for an organism.

A use of a YAP pathway activator in the preparation of an anti-hepatotropic virus medicament. In in vivo and in vitro hepatotropic virus replication models, it has been found that using the YAP pathway activator can significantly inhibit the replication of hepatotropic viruses such as hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, and hepatitis E virus and the expression of viral antigens. The antiviral effect of the activator is mainly achieved by means of regulation of the YAP signaling pathway and the fact that inhibiting YAP can restore viral replication. The described YAP pathway activator may be specifically a LATS1 / 2 kinase inhibitor, such as TDI-011536 or TRULI. The YAP pathway activator has no significant cytotoxicity and has a good antiviral effect.

The present invention discloses a pan-genotype ORF3 protein of the Hepatitis E virus genus A and an application thereof. The HEV-A pan-genotype ORF3 protein can be used to detect the content of IgG and IgM antibodies in the serum of hepatitis E patients. After the serum is diluted 500 times, IgG and IgM in the serum can still be detected, with high sensitivity. Since the ORF2 polypeptide is the main component of the HEV vaccine, the ORF2 antibody detection kit cannot distinguish between vaccine-immunized people and infected people. However, HEV-positive cases detected by the ORF3 protein detection kit can all be judged as people naturally infected with HEV.

Virus-Like Particles derived from the subfamilies, Parahepevirinae and Orthohepevirinae, particularly those of the species Paslahepevirus balayani, which can cause acute hepatitis in humans and several mammalian species, and chronic conditions in immunocompromised patients are disclosed. Compositions of VirusLike Particles comprising viral capsid proteins capable of assembly in cultured cells that may be purified, disassembled, and reassembled in the presence of other molecules suitable for use as therapeutic drug products to facilitate the targeting and delivery of cargo molecules to specific cells or tissues, or as antigenic agents designed to stimulate responses to heterologous epitopes exposed on the surfaces of Virus- Like Particles are provided. Functional capsids comprising polypeptide sequences comprising amino acid substitutions, insertions, or deletions of amino acid encoded by a consensus of ORF2 genes, that are functionally-similar or have enhanced properties compared to capsid polypeptides encoded by naturally-occurring viruses obtained from clinical samples or prototype Hepatitis E Viruses (HEV) are provided.

The invention relates to the technical field of biological detection, and discloses an oligonucleotide composition, a kit and application. The composition disclosed by the invention can be used for rapidly and simultaneously detecting salmonella, shigella, entamoeba histolytica, vibrio cholerae, hepatitis A virus and hepatitis E virus in a sample with high specificity, and has relatively high sensitivity and stability. Compared with the traditional detection methods such as pathogen separation, genome sequencing and the like, the efficiency of identifying and detecting salmonella, shigella, entamoeba histolytica, vibrio cholerae, hepatitis A virus and hepatitis E virus by adopting the composition disclosed by the invention is effectively improved, the detection process is simple and convenient to operate, and the composition is suitable for large-scale popularization and application.

The invention discloses an exogenous gene delivery method, which comprises the following steps of: transfecting vesicles by using a fusion expression vector or a co-expression vector of an ORF3 protein gene of hepatitis E virus and an exogenous gene to generate cells, purifying cell culture supernatant by using a differential ultracentrifugation method to obtain vesicles, transfecting vesicle recipient cells by using the vesicles, and transferring the vesicles to obtain the exogenous gene. And further transferring an exogenous gene product into recipient cells. The invention also discloses a gene editing method, which comprises the following steps of: preparing a carrier which contains the fusion gene of the ORF3 protein of the hepatitis E virus and the Cas9 protein, inserting independently expressed sgDNA, transfecting vesicles by using the carrier to generate cells, purifying cell culture supernatant by using a differential ultracentrifugation method to obtain the vesicles, transfecting vesicle recipient cells by using the vesicles, and obtaining the gene editing product. And target genes corresponding to the sgDNA are edited in recipient cells. The method disclosed by the invention has an application prospect.

The application provides an antigen for detecting hepatitis E virus IgM and a preparation method and application thereof, relates to the technical field of biotechnology, and comprises the following (A) or (B); (A) a protein composed of the amino acid sequence shown in SEQ ID NO. 1; (B) a protein composed of the C terminal of the amino acid sequence shown in SEQ ID NO. 2 and the N terminal of the amino acid sequence shown in SEQ ID NO. 1. It is found that the ORF2aa417-660 antigen and the ORF2aa417-660 antigen added with a beta-fold structure sequence can be used as a point membrane raw material for detecting IgM antibodies in a colloidal gold product, the positive detection rate is high, the false positive rate is low, and the sensitivity is obviously better than that of the ORF2aa459-606 antigen. In particular, the ORF2aa417-660 antigen added with the beta-fold structure sequence is helpful to the formation of a dimer and a multimer of the antigen, and further enhances the immunogenicity of the antigen.

Virus-Like Particles derived from the subfamilies, Parahepevirinae and Orthohepevirinae, particularly those of the species Paslahepevirus balayani, which can cause acute hepatitis in humans and several mammalian species, and chronic conditions in immunocompromised patients are disclosed. Compositions of VirusLike Particles comprising viral capsid proteins capable of assembly in cultured cells that may be purified, disassembled, and reassembled in the presence of other molecules suitable for use as therapeutic drug products to facilitate the targeting and delivery of cargo molecules to specific cells or tissues, or as antigenic agents designed to stimulate responses to heterologous epitopes exposed on the surfaces of Virus- Like Particles are provided. Functional capsids comprising polypeptide sequences comprising amino acid substitutions, insertions, or deletions of amino acid encoded by a consensus of ORF2 genes, that are functionally-similar or have enhanced properties compared to capsid polypeptides encoded by naturally-occurring viruses obtained from clinical samples or prototype Hepatitis E Viruses (HEV) are provided.

The application discloses a primer set for detecting a whole genome of a pig HEV, a kit comprising the primer set and application. The primer set comprises nucleotide sequences as shown in SEQ ID NO. 1-62, the primer set is designed for amplification fragments of 450 bp of a whole gene of the pig HEV and 650 bp of a non-conserved region, is divided into two sets of shingle degenerate primers, comprises five primer groups of A, B, C, D and E, and five amplification primer pools are constructed by using the five primer groups, so that the primer set can cover the complete genome of different strain types of the pig HEV. The application also establishes a method for sequencing the whole genome of the pig hepatitis E virus by using multiplex targeted RT-PCR and nanopore sequencing based on the primer set. The method is rapid, sensitive and specific, can be used for rapidly obtaining the whole genome information of the pig hepatitis E virus in a sample, and provides a reliable technical means for tracing the pig hepatitis E virus and accurately determining the genotype.

This invention relates to the field of gene detection technology, and more particularly to a capture probe combination, a kit, and its application in hepatitis E virus whole-genome sequencing. The probe combination comprises probes with nucleotide sequences as shown in SEQ ID NO.1-SEQ ID NO.951. The applications include: (1) detecting hepatitis E virus, or preparing products for detecting hepatitis E virus; (2) whole-genome sequencing of hepatitis E virus, or preparing products for whole-genome sequencing of hepatitis E virus. This invention provides a set of probes for whole-genome sequencing of hepatitis E virus, which can specifically enrich ultra-low copy number hepatitis E virus genomes from high background host nucleic acids, and thus has significant application value in hepatitis E virus detection in various scenarios such as clinical diagnosis, epidemic tracing, virus mutation monitoring, food safety testing, and basic research.

The invention discloses a primer probe combination for detecting hepatitis E virus, a method and application. The nucleotide sequence of the primer comprises sequences as shown in SEQ ID NO. 1 to SEQ ID NO. 4; the nucleic acid sequence of the fluorescent probe comprises a sequence as shown in SEQ ID NO. 5 to SEQ ID NO. 6. The primer probe combination and the detection method provided by the invention can cover each gene subtype of genotype 3 and genotype 4 hepatitis E viruses and perform typing on the genotype 3 and genotype 4 hepatitis E viruses, and are high in sensitivity and simple to operate.

The present invention discloses a hepatitis E virus ORF3 protein mutant, gene, and recombinant adenovirus. The hepatitis E virus ORF3 protein mutant of the present invention is obtained by deleting the 56-76aa segment of the hepatitis E virus ORF3 protein, and the amino acid sequence is shown in SEQ ID No. 3. The hepatitis E virus ORF3 protein mutant of the present invention is obtained by deleting the 56-76aa segment of the ORF3 amino acid sequence. This mutant loses the ability to co-localize with mitochondria but retains the immunogenicity of ORF3. When prepared as a vaccine, it can avoid the health risks caused by the co-localization of ORF3 with mitochondria.

The invention discloses a preparation method of a zinc coordination polymer and application of the zinc coordination polymer in virus resistance, and belongs to the technical field of metal organic complexes. The chemical formula of the zinc coordination polymer is {[Zn (ndba) (dib)]. H2O} n, ndba2 is obtained by losing two protons from 3, 3-(naphthalene-2, 7-diyl) dibenzoic acid (H2ndba), and dib is diimidazolyl benzene; the zinc coordination polymer material belongs to an orthorhombic system and a Pbca space group, and the cell parameters are as follows: a is equal to 12.9799 (5), b is equal to 15.8775 (6), c is equal to 28.7281 (10), alpha is equal to beta, gamma is equal to 90 degrees, and the cell volume V is equal to 5920.5 (4) 3; each tetracoordinated metal zinc ion is located in the center of the deformed tetrahedron geometrical configuration and connected with two ndba2 and two dib, a long-range ordered one-dimensional tubular structure is formed in the a-axis direction, and the long-range ordered one-dimensional tubular structure and the long-range ordered one-dimensional tubular structure are interspersed and expanded into a three-dimensional supramolecular network. The zinc coordination polymer provided by the invention can effectively inhibit the replication of hepatitis E virus, is obtained by one-step synthesis through a solvothermal method, is simple and feasible in preparation process, short in preparation period, green and environment-friendly in reaction condition and high in yield, and provides an ideal material for a novel virus inhibitor.

The invention discloses a primer group for detecting a porcine HEV whole genome, a kit comprising the primer group and application. The primer group comprises nucleotide sequences as shown in SEQ ID NO.1-62, the primer group is designed aiming at amplified fragments of 450bp of a whole gene and 650bp of a non-conserved region of the pig HEV, and is divided into two sets of imbalanobis degenerate primers, the two sets of imbalanobis degenerate primers comprise five primer groups A, B, C, D and E, five amplification primer pools are respectively constructed by the five primer groups, and the primer groups can be ensured to cover complete genomes of different plant types of viruses of the pig HEV. The invention also establishes a method for sequencing the whole genome of the porcine hepatitis E virus by multi-targeting RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and nanopore sequencing on the basis of the primer group. The method is quick, sensitive and specific, can be used for quickly acquiring the whole-genome information of the porcine hepatitis E virus in a sample, and provides a reliable technical means for tracing the source of the porcine hepatitis E virus and accurately judging the genotype.