59 results about "Adenovirus infection" patented technology

The present invention addresses the need to improve the yield of adenovirus when grown in cell culture systems. In particular, it has been demonstrated that for adenovirus, the use of infection temperatures lower than 37° C. in a cell culture system results in improved yields of adenovirus. In addition, it has been demonstrated that when host cells are grow in a bioreactor, initiating adenovirus infection by diluting the host cells with fresh media and adenovirus results in improved yield of adenovirus. Methods of adenoviral production and purification using infection temperatures less than 37° C. are disclosed. Methods of adenoviral production and purification wherein the host cells are grown in a bioreactor and adenovirus infection is initiated by diluting the host cells with fresh media and adenovirus are also disclosed.

The invention relates to preparation and application of a gene chip capable of detecting adenoviruses in a parting mode.A preparation method comprises the steps of preparing specific primers and probes for different types of adenoviruses, preparing oligonucleotides chips, establishing a plurality of PCR systems, and establishing a hybridization system.The gene chip prepared through the method can screen different types of adenoviruses including a 3-type adenovirus, a 7-type adenovirus, a 14-type adenovirus, an 11-type adenovirus and a 55-type adenovirus.The gene chip has the advantages of quickness, accuracy, high throughput and high specificity.A new detection means is provided for clinical diagnoses of different types of adenovirus infections and epidemiological surveys.

The present invention provides methods and compositions for conferring tumor selective cell death on cancer cells expressing specific target precursor messenger RNA molecules (cancer cell selective target pre-mRNAs). The compositions of the invention include conditionally replicative adenoviruses that have been genetically engineered to express one or more pre-trans-splicing molecules (PTMs) designed to interact with one or more cancer cell target pre-mRNA and mediate a trans-splicing reaction resulting in the generation of novel chimeric RNA molecules (chimeric RNA) capable of encoding adenovirus specific protein(s). Adenovirus specific proteins include those proteins complementing an essential activity necessary for replication of a defective adenovirus. The methods and compositions of the invention may be used to target a lytic adenovirus infection to cancer cells thereby providing a method for selective destruction of cancer cells. In addition, the adenoviruses of the invention may be engineered to encode PTMs designed to interact with target pre-mRNAs encoded by infectious agents within a cell, thereby targeting selective destruction of cells infected with such agents.

A method for the prevention or treatment of Influenza virus infection or Adenovirus infection by administering an effective amount of a compound of Formula (I), Formula (II), or similar compound to an individual in need is provided.

The invention belongs to the technical field of virus detection and provides an extracting solution for extracting human adenovirus DNA (deoxyribonucleic acid), a PCR (polymerase chain reaction) reaction solution for detecting human adenoviruses, a PCR primer sequence for detecting human adenovirus DNA, a rapid detection kit for human adenoviruses and a method for quantitatively detecting the quantity of human adenoviruses. The rapid detection kit for human adenoviruses comprises the extracting solution for extracting human adenovirus DNA, the PCR reaction solution for detecting human adenoviruses and the PCR primer sequence for detecting human adenovirus DNA, and further comprises an adenovirus positive control and an amplification primer sequence for detecting the positive control. The adenovirus detection sensitivity of the kit disclosed by the invention can reach 500copies/ml. The quantitative linear range is 500-5.0E+08copies/ml, so that the kit is suitable for detecting the adenoviruses in a trace sample and provides a reliable basis for early diagram of adenovirus infection.

The invention discloses three neutralizing epitope of human adenovirus type 3 (HAdV-3) and type 7 (HAdV-7) and an application thereof. The amino acid sequence of the neutralizing epitope of the HAdV-3 and HAdV-7 is selected from the amino acid sequence shown in SEQ ID NO: 1, 2, 3, and the nucleotide sequence of the neutralizing epitope of HAdV-3 and HAdV-7 is selected from the nucleotide sequence shown in the SEQ ID NO: 4, 5, 6. The neutralizing epitopes of HAdV-3 and HAdV-7 can be used for preparing vaccines or antibody and antigen binding fragments for preventing infection of human adenovirus type 3 and human adenovirus type 7, and the antibody and the antigen binding fragments can be used for preparing drugs for preventing or curing adenovirus infection. The drugs contain the antibody or antigen binding fragments. The invention also provides a method for preventing and curing adenovirus infection, i.e. the vaccines or drugs with immune effective quantity are applied. The three neutralized epitopes of HAdV-3 and HAdV-7 can be served as target protein for developing a diagnostic kit.

The invention discloses a fluorescence quantitative PCR kit for detecting type-3 cow adenovirus and application. The kit comprises a) DNA extraction reagent, b) hot starting Taq DNA polymerase, c) primers and TaqMan probe, d) standard positive DNA template, and e) PCR fluorescence quantitative reaction liquid. The kit is characterized in that the sequence of a positive primer is 5'-CCTGAATTCTCTTGCAGCCAGA-3', the sequence of a negative primer is 5'-CCTACCGAACCGACGCAGAT-3', the size of an amplicon is 100bp, the sequence of a fluorescence probe is 5'-FAM-TGAGAAGGTACTCCTCGTCGCTGGACCA-TAMRA-3', a 5' end of the probe marks a fluorescence emitting group FAM, a near 3' end of the probe marks a fluorescence quenching group TAMRA, the standard positive DNA template converts colon bacillus DH5a by a pGEM-T carrier inserted into a type-3 adenovirus pol protein 100bp fragment, plasmids are extracted after multiplication to prepare the kit, and A260 is measured by an ultraviolet spectrophotometer to definite quantity and is diluted by 10 times of gradient. The kit efficiently and conveniently monitors type-3 cow adenovirus pollution in a serum product in real time, can be widely applied to epidemiology research on adenovirus infection, can provide technical support for related basic research, and has wide application prospect.

The present invention provides methods and reagents for detecting antibodies to adenovirus. In a preferred embodiment, the present invention is useful for detecting antibodies to adenovirus serotype 5. In a further preferred embodiment, the present invention can be used in a biosensor-based assay. It is contemplated that the present invention is useful to detect antibodies for ascertaining adenovirus infection, evaluating patient response to gene therapy using adenovirus vectors, developing vaccines to adenovirus infection, developing therapeutics for inducing passive immunity to adenovirus infection, as well as other uses.

The invention relates to a production method of a recombinant adenovirus gene vaccine for preventing the novel coronavirus. The production method comprises the following main steps: amplifying cells;performing inoculating in a bioreactor, carrying out perfusion culturing in a culture solution, and carrying out cell re-amplification; diluting the cells and transferring the processed cells to a large-volume bioreactor; infecting and amplifying the recombinant adenovirus; harvesting the recombinant adenovirus culture solution and then performing filtering and concentrating; performing nuclease treatment; carrying out full-automatic ion exchange column chromatography separation and purification; and preparing, filling and freeze-drying the product. The method provided by the invention has good amplification and economy, can meet various technical requirements of medical monitoring organizations on GMP production, and fully ensures the quality and biological activity of the product.

The invention relates to a lymphocyte culture method, and in particular discloses a culturing method of adenovirus gene-modifying tumor specificity cytotoxic T lymphocyte. The method comprises: taking 150-200ml of bleeding of the umbilicus or autologous peripheral blood for single karyocyte separation, separately culturing DC cells by adhering to a wall for 2h, and performing T lymphocyte preculture on the non-adhered cells in an IL-2-contained culture medium; collecting mature DC after the DC cells are mature through induction of iAPA factor adenovirus infection, stimulating the autologous primary T lymphocyte to differentiate into cytotoxic T lymphocyte according to an effect target ratio of DC to T being 1 to 10, and observing and recording multiplication capacity of CTL and detecting the cytotoxicity of the CTL. The limitation of a blood component single sampling machine is solved, collected blood volume is less, burden of a patient is reduced, and iAPA-DC can be observed in vitro to stimulate and differentiate the primary T lymphocyte into tumor specificity cytotoxic T lymphocyte.

The invention discloses a recombinant adenovirus for expressing African swine fever virus B602L-B646L protein and a construction method thereof, and belongs to the technical field of genetic engineering. A recombinant adenovirus shuttle vector pENTRE-EGFP-TOPO is utilized, and a recombinant adenovirus vector pAD-CMV-EGFP-B602L-B646L is obtained through a series of intermediate processes; according to the present invention, the African swine fever virus B602L-B646L protein expression recombinant adenovirus is constructed, the African swine fever virus B602L-B646L protein expression recombinant adenovirus is linearized and transfected with AD293 cells, the recombinant virus is screened according to the cytopathic effect formed by adenovirus infection, the adenovirus packaging process is achieved, the recombinant adenovirus expressing the African swine fever virus B602L-B646L protein is obtained, and the foundation is laid for the construction of the recombinant adenovirus vaccine expressing the African swine fever virus

The invention discloses a high-throughput screening method and application of a drug for inhibiting human adenovirus proliferation. A human adenovirus marked with GFP in the patent can express green fluorescent protein after infecting with HEK-293 cells, so that the infected cells are green under a fluorescence microscope. The anti-human adenovirus drug activity is realized through the following steps: detecting the change of the adenovirus content in the supernatant of a culture medium after the anti-adenovirus drug and the human adenovirus are added into the HEK-293 cells by using flow cytometry. The specific operation comprises the following steps: taking a proper amount of the virus supernatant, adding the virus supernatant into a certain quantity of the HEK-293 cells again, and determining the ratio (cells with green fluorescence) of the adenovirus infected cells in the cells by flow cytometry so as to evaluate whether the added drug has an obvious inhibition effect on adenovirusproliferation or not. According to the method, whether candidate drug molecules have the effect of inhibiting human adenoviruses or not can be screened quickly, simply, conveniently and effectively invitro in a low-cost and high-throughput manner. According to the method, the research and development of novel anti-human adenovirus drugs can be accelerated.

The invention is about a method that can produce recombination adenovirus vector highly, and belongs to the field of cell engineering technology. The cell, which puts the HEK cell as the recombination adenovirus vector, can produce the dissociative cell into cell mass under the Suspension Culture, or depolymerization cell mass as dissociative cell, using the free settling of cell mass or rotary filter interception cell mass and replacing substrate or affusing substrate continuously, it can recombinate the adenovirus infection HEK cell mass and culture it to amplify in the cell. Its advantages are:óƒThe activity of the cell is greater or equal to 90%, the consistency of live cell is greater or equal to 1í‡10 to the power 7 cells/ml.óIt uses the recombination adenovirus vector to infect HEK cell mass directly and enlarge breed in the cell, and can carry into execution substrate replacing or continuous perfusion with the free settling of HEK cell mass or intraoral rotary filter.ó�The titer of pAd-TH-GFP can reach 1-5í‡10 to the power 13 GTU/ml, the product capacity of 5L stirred tank bioreactor can reach 1-2í‡10 to the power 17 GTU/ml, the gain of recombination adenovirus vector is convenient.óœThe producing process is tight and unhindered, and the size can be enlarged easily.

The invention relates to the technical field of genetic engineering vaccines, and particularly discloses an I-group 4-type fowl adenovirus fiber-2 protein antigen as well as a method for preparing a genetic engineering subunit vaccine and application of the I-group 4-type fowl adenovirus fiber-2 protein antigen. The method comprises the following steps: adding a non-coding gene in front of a fiber-2 gene of the fowl adenovirus FAdV-4, cloning the non-coding gene to a prokaryotic expression vector pET-28a, transforming the prokaryotic expression vector pET-28a to escherichia coli to construct an engineering bacterium, and inducing the engineering bacterium to express to obtain the soluble fiber-2 protein. Wherein the amino acid sequence of the aviadenovirus fiber-2 antigen protein is SEQ ID NO.2, the nucleotide sequence of a gene coded by the aviadenovirus fiber-2 antigen protein is SEQ ID NO.1, and soluble protein with high expression quantity is obtained through non-coding gene regulation and control. The fiber-2 protein of the fowl adenovirus is expressed through escherichia coli, a soluble expression product is obtained, the subunit vaccine is prepared, safety and effectiveness are achieved, and fowl I-group 4-type adenovirus infection can be prevented.

The invention relates to a method for detecting type-5, type-7 and type-55 adenoviruses by using a mass spectrum multiple reaction monitoring technology. The method comprises the following specific steps: synthesizing specific peptide fragment genes of hexon proteins of type-5, type-7 and type-55 adenoviruses; preparing a labeled peptide fragment mixed reagent; determining an ion peak value of the diagnostic marker; preparing the kit; and detecting the sample. The invention provides a new marker for early warning and diagnosis of adenovirus type, type 7 and type 55 infection, and has important clinical value. A mass spectrum result is utilized to confirm that mass spectrum ion peaks consistent with the standard and the heavy standard of preset adenovirus type 5, type 7 and type 55 hexon protein symbolic peptide fragments appear in a serum sample of a patient in the early stage of adenovirus infection. The SILAC-labeled winning and heavy-labeled hexon protein standard mixed reagent of adenovirus type 5, type 7 and type 55 can be used as an early warning and diagnosis marker for adenovirus type, type 7 and type 55 infection, and the result is accurate.

The invention discloses a recombinant adenovirus for expressing African swine fever virus EP153R-EP402R protein and a construction method of the recombinant adenovirus, and belongs to the technical field of genetic engineering. A recombinant adenovirus shuttle vector pENTRE-EGFP-TOPO is utilized, a recombinant adenovirus vector pAD-CMV-EGFP-EP153R-EP402R is obtained through a series of intermediate processes, and the recombinant adenovirus vector is linearized to transfect AD293 cells, the recombinant virus is screened according to the cytopathy formed by adenovirus infection, and the adenovirus packaging process is achieved, the recombinant adenovirus for expressing African swine fever virus EP153R-EP402R protein is obtained, and the foundation is laid for the construction of the African swine fever virus EP153R-EP402R protein expression.

The invention discloses a recombinant adenovirus for expressing African swine fever virus EP153R protein and a construction method thereof, and belongs to the technical field of genetic engineering, the recombinant adenovirus shuttle vector pENTRE-EGFP-TOPO is utilized, and a recombinant adenovirus vector pAD-CMV-EGFP-EP153R is obtained through a series of intermediate processes, and after being linearized, the recombinant adenovirus vector pAD-CMV-EGFP-EP153R is transfected into an AD293 cell; the recombinant virus is screened according to cytopathy formed by adenovirus infection, the adenovirus packaging process is realized, the recombinant adenovirus for expressing the African swine fever virus EP153R protein is obtained, and a foundation is laid for constructing a recombinant adenovirus vaccine for expressing the African swine fever virus EP153R protein.

The invention belongs to the technical field of medicines, and particularly relates to application of VER-49009 in preparation of a medicine for preventing and/or treating adenovirus infection. The invention discloses an application of VER-49009 in preparation of a medicine for inhibiting adenovirus and/or a medicine for preventing and/or treating adenovirus infection, so that a safe and effective small molecular compound is provided for clinically treating the adenovirus. The VER-49009 can effectively inhibit the replication of the adenovirus in a non-toxic range, can be further developed into a medicine for treating or preventing diseases caused by adenovirus infection, and has a wide application prospect.