70 results about "Adenovirus typing" patented technology

A packaging cell line capable of complementing recombinant adenoviruses based on serotypes from subgroup B, preferably adenovirus type 35. The cell line is preferably derived from primary, diploid human cells (e.g., primary human retinoblasts, primary human embryonic kidney cells and primary human amniocytes) which are transformed by adenovirus E1 sequences either operatively linked on one DNA molecule or located on two separate DNA molecules, the sequences being operatively linked to regulatory sequences enabling transcription and translation of encoded proteins. Also disclosed is a cell line derived from PER.C6 (ECACC deposit number 96022940), which cell expresses functional Ad35 E1B sequences. The Ad35-E1B sequences are driven by the E1B promoter or a heterologous promoter and terminated by a heterologous poly-adenylation signal. The new cell lines are useful for producing recombinant adenoviruses designed for gene therapy and vaccination. The cell line can also be used for producing human recombinant therapeutic proteins such as human growth factors and human antibodies. In addition, the cell lines are useful for producing human viruses other than adenovirus such as influenza virus, herpes simplex virus, rotavirus, measles virus.

Adenovirus types 11p and 4p show a higher binding affinity and infectivity than type 5 for endothelial and carcinoma cell lines. Adenovirus type 11p shows a stronger binding to cells for neural origin, such as glioblastoma, neuroblastoma and medulloblastoma. The fact that adenovirus type 11 has a comparatively low prevalence in society, together with its high affinity and infectivity, makes it very suitable for use in gene therapy.

Adenovirus types 11p and 4p show a higher binding affinity and infectivity than type 5 for endothelial and carcinoma cell lines. Adenovirus type 11p shows a stronger binding to cells for neural origin, such as glioblastoma, neuroblastoma and medulloblastoma. The fact that adenovirus type 11 has a comparatively low prevalence in society, together with its high affinity and infectivity, makes it very suitable for use in gene therapy.

The invention belongs to the field of antiviral drugs. The invention provides applications of Gramine and derivatives thereof to preparation of medicaments for resisting adenovirus Type 7. Gramine derivatives are compounds shown in G1,G2,G3,G4,G5,G10,G17,G18,G19,G20, and G21. Experiments for researching activity of Gramine derivatives for resisting ADV7, pharmacology research of Gramine derivatives for resisting ADV7, and tests of inhibition effects of G19 for ADV7 duplication show that Gramine derivatives can inhibit cytopathic effects (CPE) generated by ADV7 on host cell Hela, enhance cell survival rate, inhibit yield of progeny virus, and mainly inhibit the duplication phase of ADV7 virus in host cells. The Gramine and derivatives thereof have potential for preparing anti-ADV1 virus medicaments, and the compounds have the advantages of simple synthesis process, and economy and fastness; the compounds are easy for large-scale production, and have clinic application prospects.

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 invention provides an inactivated vaccine strain CH-GD-12-2014 against the duck adenovirus type 3. The inactivated vaccine strain CH-GD-12-2014 is preserved in China Center for Type Culture Collection, Wuhan University, Wuhan City, China, on November 27, 2017, with an accession number of CCTCC No. V201764. The whole gene sequence of the strain is submitted to Genebank, with an accession numberof KR135164. The invention also provides a polypeptide with an amino acid sequence as shown in SEQ ID No. 2, and a nucleotide sequence encoding the polypeptide, wherein the nucleotide sequence is asshown in SEQ ID No. 1. An inactivated strain provided by the invention is safe to all kinds of ducks and free of side reactions. The vaccine prepared from the inactivated strain is safe and effective,can prevent attacks caused by homologous virulent viruses, and has practical and good application value.

A packaging cell line that complements recombinant adenoviruses based on serotypes from subgroup B, preferably adenovirus type 35. The cell line is preferably derived from primary, diploid human cells that are transformed by adenovirus E1 sequences either operatively linked on one DNA molecule or located on two separate DNA molecules, the sequences being operatively linked to regulatory sequences enabling transcription and translation of encoded proteins. Also disclosed is a cell line derived from PER.C6 that expresses functional Ad35 E1B sequences. The Ad35-E1B sequences are driven by the E1B promoter or a heterologous promoter and terminated by a heterologous poly-adenylation signal. The cell lines are useful for producing recombinant adenoviruses designed for gene therapy and vaccination. The cell lines can also be used for producing human recombinant therapeutic proteins such as human growth factors and human antibodies. Also, the cell lines are useful for producing human viruses other than adenovirus such as influenza virus, herpes simplex virus, rotavirus, and measles virus.

The invention aims at providing a duck adenovirus type 2 inactivated vaccine, wherein an antigen is an inactivated GD strain virus; and the GD strain virus is preserved in China Center for Type Culture Collection in Wuhan University on June 5, 2016 with preservation number of CCTCC No: V201633. The duck adenovirus type 2 inactivated vaccine prepared by the invention is good in safety and free from any local and systemic adverse reactions caused by the vaccine. Based upon analysis on characters, safety test and efficacy test data in a preservation period test, various indexes are stable and effective; and a result of assessing an immune effect of the vaccine by virtue of a serological method and an immune attack method shows that the inactivated vaccine prepared by the invention can achieve effective immune protection on ducks, and the inactivated vaccine has a good commercial development prospect.

The invention discloses a duck adenovirus type-3 virus strain and yolk antibody preparation and application thereof. The duck adenovirus type-3 virus strain is produced by a duck adenovirus type-3 virus GD06 strain immune poultry with the preservation number being CCTCC NO:V201905. The virus strain has the good specificity and immunogenicity. The yolk antibody of the adenovirus type-3 is high in safety, infection of the duck adenovirus type-3 is effectively prevented, the immunity protecting rate is high, and the duck adenovirus type-3 is effectively treated; the virus strain can be used for detecting duck adenovirus type-3 virus and has the wide application prospect.

The invention discloses a PCR primer for detecting duck adenovirus type 4 as well as a detection method and application thereof. The primer capable of rapidly detecting the duck adenovirus type 4 andthe PCR detection method of the primer are established for the first time, operation is easy, cost is low, and consumed time is short. Moreover, the PCR detection method for detecting the duck adenovirus type 4 is high in accuracy, good in specificity and high in sensitivity, the lowest detection limit is 6*10 <1> copies, clinical popularization and application are facilitated, and technical support is provided for carrying out adenovirus type 4 molecular epidemic virology investigation and related detection work in duck flocks.

The invention relates to a conditionally replicating oncolytic adenoviral vector used for expressing two exogenous genes and modified by small peptide. Specifically, base 1083bp is deleted in the section of 2245bp-3327bp of human adenovirus type 5 genome, and an expression element expressing a first exogenous gene is inserted into the section of 2245bp-3327bp of human adenovirus type 5 genome. And at 32679bp of adenovirus type 5 genome, i.e. at the position of fibrin HI loop, a code containing a small peptide is inserted. A second exogenous gene of dual expression and an eGFP expression element are inserted into the section of 32787bp-32788bp of human adenovirus type 5 genome. The construction method of the vector includes the steps of: constructing a shuttle missed by pAd5E1B 55kd, constructing a shuttle of the first exogenous gene pHE1B55D-, constructing an adenoviral vector backbone of the first exogenous gene pHE1B55D- / SwaI, constructing a shuttle of the small peptide sequence pshuttle Ad5-E4-fiber-, constructing a shuttle expressing eGFP and the second exogenous gene, and preparing the conditionally replicating oncolytic adenoviral vector used for expressing two exogenous genes and modified by small peptide. The invention also provides the application of the adenoviral vector provided in the invention in preparing medicaments for treating tumours.

The invention discloses primers and a probe for rapidly and quantitatively detecting duck adenovirus type 4 and a detection method and application of the primers and the probe. The sequences of the primers are as shown in SEQ ID NO.1-2, and the sequence of the probe is as shown in SEQ ID NO.3. The fluorescent PCR method capable of rapidly and quantitatively detecting the duck adenovirus type 4 inthe clinical sample is established for the first time; the detection method is simple to operate, the whole operation process does not exceed 3 hours, the sensitivity is high, the specificity is good,the repeatability is good, quantitative analysis can be accurately and rapidly carried out with high flux, and the detection method is beneficial to popularization and application in clinical practice.

An object of the present invention is to provide a conditionally replicating adenovirus having a strong anticancer effect. A conditionally replicating adenovirus to replicate specifically in a cancer cell and express REIC protein or REIC C domain protein, wherein the conditionally replicating adenovirus is obtained by inserting full-length REIC DNA or REIC C domain DNA into a conditionally replicating adenovirus comprising an ITR (inverted terminal repeat) sequence of an adenovirus type 5 genome and insertion of an HRE sequence, an hTERT promoter, a decorin-encoding DNA, and a DNA encoding a peptide comprising an RGD sequence.

The invention relates to a selecting method for duck adenovirus 1 type virus replicating non-necessary section segment and the recombination fowl adenovirus gained through transporter gene and the universal transporter gene. It uses duck adenovirus 1 type virus as material to expand the sequence at right side of E4 area by PCR method. Inducing loxP sequence to the two sides of GFP gene expression box and inserting into the expanded segment, the universal transporter gene would be constructed. The transporter gene would gain the recombination adenovirus of stable expression GFP, thus, a replicating non-necessary area would be determined. The universal transporter gene loxP-GFP-loxP sequence side contains a unique enzyme Restriction Enzyme cutting site, and cold insert relative target gene to gain recombination virus without GFP gene. The gene engineer medicine developed from the universal transporter gene would not contain selecting marker gene.

The embodiment of the invention relates to the field of in-vitro detection, in particular to a kit for rapidly and accurately detecting adenovirus type 3 and a preparation method of the kit. The kit provided by the embodiment of the invention comprises an IgM antibody enzyme-labeled working solution, an IgA antibody enzyme-labeled working solution and an IgG antibody enzyme-labeled working solution. The IgG antibody enzyme-labeled working solution comprises a peroxidase-labeled anti-human IgG antibody and an ELISA plate coated with an adenovirus 3 type recombinant antigen; the adenovirus 3 type recombinant antigen comprises one or more of a recombinant antigen A, a recombinant antigen B or a recombinant antigen C. The kit (enzyme linked immunosorbent assay) provided by the invention makesup the blank in the market;, the detection results are comprehensively considered by respectively detecting adenovirus type 3 IgM, IgG and IgA antibodies in human serum or blood plasma, the detectionresults are complementary with each other, the reliability is high, the specificity is good, and the sensitivity is high; the kit can be complementary with nucleic acid detection, and is suitable forclinical diagnosis and epidemiological investigation.

The invention provides an amplification method of internal reference containing double isothermal nucleic acid for rapidly detecting type-7 adenovirus and belongs to the technical field of isothermalnucleic acid amplification. According to the amplification method, a specific primer pair and a probe for HAdV-7 detection are designed first through human adenovirus type-7 (HAdV-7) gene sequencing and alignment, an internal reference probe is designed, and the nucleotide sequences of the specific primer pair and the probe are respectively as shown in SEQ ID NO.1-4, an internal reference containing double isothermal nucleic acid amplification system is established, and a kit for detecting adenovirus type 7 is further constructed. The method is carried out under isothermal conditions, the amplification of the type-7 adenovirus and the internal reference DNA can be realized within 5-20min, the sensitivity is high, the specificity is good, false negative and invalid results are eliminated due to the addition of an internal reference, the method is more suitable for the detection of a large number of samples, the clinical application is facilitated, and the method is suitable for the rapid detection of the type-7 adenovirus.

The invention provides a kind of recombinant adenovirus expressing human granulysin, which genome is deleted E1 and E3, and inserted the expression cassette of human granulysin in E1, and the expression cassette includes: promoter sequence, human granulysin coding sequence and polyadenylation signal by turns. The adenovirus type is Ad5. The preparation method includes: a. cloning the gene of human granulysin, b. constructing the plasmid pAd-GRA expressing human adenovirus, c. digesting pAd-GRA with enzyme PacI, conventionally extracting with phenol, chloroform / isoamyl alcohol, washing with ethanol, suspending in TE 50 mu l under sterile conditions, and transferring -293 cells.

The invention discloses an Ad4 / Ad7 type bivalent recombinant adenovirus and application thereof. The invention provides the recombinant adenovirus, which is characterized in that compared with the genomic DNA of a human type 4 adenovirus, the genomic DNA of the recombinant adenovirus only undergoes the following three mutations: 1, the first to eighth nucleotides mutate into 'CATCATCA' from 'ctatctat'; 2, the 35983th-35990th nucleotides mutate into 'TGATGATG' from 'atagatag'; 3, replacing DNA molecules for encoding a fifth hypervariable region of an adenovirus type 4 hexon with DNA molecules for encoding a fifth hypervariable region of an adenovirus type 7 hexon. The invention has a great application value for the prevention and control of Ad4 adenovirus and Ad7 adenovirus.

The invention discloses a primer probe combination, a kit and a detection method for detecting duck adenovirus type 3 based on an RAA technology. The primer probe combination is designed according toduck adenovirus type 3 virus specific Hexon gene, and the kit and the detection method utilize the primer probe combination to perform RAA reaction on a to-be-detected sample. Whether the to-be-detected sample contains the duck adenovirus type 3 virus or not is judged according to an amplification reaction result. The kit and the detection method are applied to rapid detection of the duck adenovirus type 3 virus, and accurate, rapid, sensitive and convenient detection of the duck adenovirus type 3 virus is realized.

The invention belongs to the technical field of veterinary biological products, and discloses a fused gene. A base sequence is as shown in a sequence table SEQ ID NO: 1. An avian influenza H9 subtypeHA gene, an adenovirus type 4 fiber2 gene and a specific connecting peptide segment gene are fused to form the fused gene. An expression protein antigen corresponding to the fused gene can induce poultry to generate high-level specific antibodies, and the poultry is protected from infection of avian influenza and avian adenoviruses. Besides, the invention further discloses a recombinant expressionvector, an antigen, a preparation method and an application.

The invention discloses a metagenome-based human adenovirus molecular typing and tracing method and system, and the method comprises the steps: constructing a human adenovirus typing database comprising a typing hierarchical classification system, and obtaining a classification unit and a complete genome / marker gene sequence of human adenoviruses, determining a complete genome / marker gene sequence of each classification unit of the human adenovirus, obtaining metagenome sequencing original data of a clinical sample, preprocessing the metagenome sequencing original data to obtain target metagenome data, analyzing the target metagenome data by utilizing a preset metagenome sequencing data dual-comparison annotation system, determining whether the clinical sample has a target human adenovirus classification unit or not, and when it is determined that the clinical sample has the target human adenovirus classification unit, determining a target associated feature corresponding to the target human adenovirus classification unit, and generating a human adenovirus typing identification report. Classification unit (subtype / genotype) typing identification of human adenoviruses can be carried out on clinical infection samples with low virus content.

“Pharmacological enhancement and manufacturing method of the antiviral compound” can be categorized into the combined patent of the pharmacological activity as well as method of isolating and purifying the naturing material. Our product adopts 9 medicinal material which are processed strictly. The stable and high quality is ensured by the WLD resin adsorption and gas chromatography. The antiviral prevention and treatment is the most urgent but unsolved problem before the American doctors. The antiviral compound has an unexpected effect on a broad spectrum of viruses including RSV, Adenovirus type 3, Influenza A1 and A3. The antiviral effect on mouse Influenza A1 is very obvious. The definite efficacy in the treatment of acute pharyngitis and tonsillitis have been proved by the clinical trial which showed the total effective rate in the above two disease were 92.3% and 87.5% respectively.

The invention discloses a Muscovy duck novel adenovirus strain, bivalent inactivated vaccine and a preparation method thereof. The bivalent inactivated vaccine is prepared by taking duck adenovirus and DAdV-3-GD MM virus as inactivated antigens. The Muscovy duck adenovirus type II and type III bivalent inactivated vaccine prepared from the screened virus has high safety, and any local and systemicadverse reaction caused by the vaccine does not appear. In the storage life experiment, each index is stable and effective through analysis of character, safety experiment and efficacy experiment data; and through evaluation on the immune effect of the vaccine by a serology method and a immune poison attack method, the Muscovy duck adenovirus type II and type III bivalent inactivated vaccine canprovide effective immune protection on Muscovy ducks and has good commercial development prospect.

The invention aims at providing a duck adenovirus type 2, wherein the duck adenovirus type 2 is a duck adenovirus type 2 GD strain which is preserved in China Center for Type Culture Collection in Wuhan University on June 5, 2016 with preservation number of CCTCC No: V201633. The duck adenovirus type 2 prepared by the invention can be used for preparing a vaccine for preventing adenovirus type 2 diseases in a group of ducks. The duck adenovirus type 2 inactivated vaccine prepared by the virus screened by the invention is good in safety and free from any local and systemic adverse reactions caused by the vaccine. Based upon analysis on characters, safety test and efficacy test data in a preservation period test, various indexes are stable and effective; and a result of assessing an immune effect of the vaccine by virtue of a serological method and an immune attack method shows that the inactivated vaccine prepared by the invention can achieve effective immune protection on the ducks, and the inactivated vaccine has a good commercial development prospect.

Disclosed is an effective and inexpensive anti-HIV agent. The anti-HIV agent comprises a combination of a recombinant viral vector having the structural gene of HIV inserted into a chimeric virus and a recombinant viral vector having the structural gene of HIV inserted into a modified vaccinia virus Ankara, wherein the chimeric virus is prepared by substituting a fiber in an adenovirus type-5 by a fiber derived from an adenovirus type-35.

The invention belongs to the field of protein engineering, and particularly relates to duck adenovirus type I Penton protein as well as a preparation method and application thereof. The inventor, by analysis, finds that the Penton protein has the advantages of hydrophilicity, no transmembrane structural domain, no signal peptide prediction and the like, and full-length expression is selected. A Penton target fragment is obtained through artificial synthesis after sequence optimization, and the expression mode of the protein is cell-free protein expression. Compared with truncated expression ofantigen dominant epitopes of traditional Penton protein, the artificially synthesized and expressed Penton recombinant protein is high in accuracy, the neutrality of the protein is guaranteed, the trouble of high mutation rate is avoided, and the application to the development of an antibody detection technology is better facilitated. In addition, an expression system has unique advantages of saving time and improving the total yield of soluble full-length protein. Furthermore, the expression system is low in cost, large in expression quantity and high in Western Blot detection reactogenicity.

The invention discloses a duck adenovirus type 3 strain, a duck adenovirus egg yolk antibody as well as a preparation method and application of the duck adenovirus type 3 strain and the duck adenovirus egg yolk antibody. The duck adenovirus type 3 strain is a duck adenovirus type 3 YJ strain and is preserved in the China General Microbiological Culture Collection Center on November 18, 2020, and the preservation number of the duck adenovirus type 3 strain is CGMCC No.21093; and the nucleotide sequence of the HEXON gene of the duck adenovirus 3 type YJ strain is as shown in SEQ ID NO. 1. The adenovirus 3 type YJ strain is obtained through separation, and the egg yolk antibody which can be used for treating the duck adenovirus 3 type virus and is efficient, safe and free of toxic and side effects is prepared.

The invention provides a probe and kit for detecting human adenovirus 7 and a use method of the kit. The kit is a human adenovirus 7 type nucleic acid detection kit based on an MIRA fluorescence method. The primer of the MIRA is designed according to the specific sequence of the hexon gene of the adenovirus type 7. The nucleic acid detection kit is rapid, specific and sensitive and does not need complex instruments and equipment.

The invention discloses a capillary-electrophoresis-based adenovirus typing detection kit and a use method therefor. The kit comprises a PCR primer group; and the PCR primer group comprises 12 kinds of adenovirus serotype forward and reverse primers, total adenovirus forward and reverse primers, human genome internal-reference forward and reverse primers and PCR internal-reference forward and reverse primers. The use method for the kit comprises the following steps: (1) pretreating a sputum or throat swab sample, and then, extracting nucleic acid from the sample; (2) carrying out multiplex PCR amplification on the extracted nucleic acid; (3) separating a PCR product by employing a capillary electrophoresis method; and (4) analyzing and reading a result. According to the kit and the use method therefor, 12 kinds of adenovirus serotypes are subjected to typing detection rapidly through combining multiplex PCR with a capillary electrophoresis analysis method. The kit has the advantages of high sensitivity, high specificity, high flux and low cost and can be used for rapidly and synchronously detecting a variety of adenovirus serotypes, and defects of the traditional methods are overcome.