29 results about "Adenovirus typing" patented technology

The present invention belongs to the field of virus detection technology, and specifically relates to a combination and application of monoclonal antibodies for the specific detection of type 5 adenovirus. The antibody combination includes 8B9 and 4A6, which target different epitopes of the HADV‑5 Hexon protein, respectively, and are suitable for a double-antibody sandwich ELISA detection system. Among them, 8B9 is used as a coating antibody, and 4A6 is used as an HRP-labeled detection antibody. The combination of the two can achieve high sensitivity and high specificity detection of HADV‑5 and replication-deficient recombinant type 5 adenovirus, and does not cross-react with other common adenovirus types. The present invention also provides the variable region amino acid sequence and nucleotide sequence of the above-mentioned antibody, as well as an ELISA detection method based on the antibody combination. The method is simple to operate, has good repeatability, and is suitable for the detection of type 5 adenovirus.

The invention provides an anti-duck adenovirus 3 type monoclonal antibody, a hybridoma cell strain and application thereof, the anti-duck adenovirus 3 type monoclonal antibody is secreted by a hybridoma cell strain 2F12, the classification name of the hybridoma cell strain 2F12 is Hybridoma cell line 2F12, the hybridoma cell strain 2F12 is preserved in China Center for Type Culture Collection on November 13, 2025, the preservation address is Wuhan University, Wuhan, China, and the preservation number is CGMCC NO. The preservation number of the strain is CCTCC (China Center For Type Culture Collection) NO: The blocking ELISA kit for detecting the duck adenovirus 3 antibody, which is prepared by utilizing the monoclonal antibody, is strong in specificity, high in sensitivity and good in repeatability, and has a very good application prospect in the aspects of epidemiological investigation of the duck adenovirus 3, vaccine immune efficacy evaluation and diagnosis of the duck adenovirus 3 disease.

The invention discloses a method, a primer group and a kit for sequencing an adenovirus whole genome, and belongs to the technical field of DNA (Deoxyribose Nucleic Acid) detection. The method for sequencing the whole genome of the adenovirus comprises the following steps: S1, carrying out a first round of amplification by adopting a first round of amplification primers with sequences as shown in SEQ ID NO.1-SEQ ID NO.93 in a table 1; and S2, carrying out a second round of amplification by adopting the second round of amplification primers with the sequences as shown in SEQ ID NO.94-SEQ ID NO.285 in the table 2. The average length of data sequenced by the method is 0.8 kb, compared with the short read length of a second-generation platform, the uniformity of adenovirus coverage can be greatly improved, meanwhile, the adenovirus type coverage is comprehensive, and the method can be widely popularized.

The present invention relates to an anti-human adenovirus type 4 Hexon protein nanobody and an application thereof, the nanobody having a unique CDR region, capable of binding to the adenovirus type 4 Hexon protein with high affinity, and capable of neutralizing adenovirus type 4 pseudovirus with high neutralizing activity in a virus neutralization test. The invention also provides application of the nano antibody in preparation of a medicine for treating and / or preventing human type 4 adenovirus infectious diseases and a diagnostic kit.

This invention relates to the quantitative detection of pigeon adenovirus and the simultaneous differential diagnosis of type I and type II. Specific primers and probes for pigeon adenovirus type I and II are designed using specific gene sequences. Real-time quantitative PCR can accurately detect and differentiate between pigeon adenovirus types I and II. Verification has shown that this technology can rapidly and accurately differentiate between PiAdV-I and PiAdV-II, exhibiting high specificity and sensitivity. Furthermore, it is cost-effective, easy to operate, and suitable for import / export quarantine and specialized laboratories for the detection and differentiation of PiAdV-I and PiAdV-II.

The invention discloses a pigeon adenovirus type 1 strain YJ-NM and application thereof, and belongs to the technical field of bioengineering. The pigeon adenovirus 1 type strain YJ-NM provided by the invention is preserved in China Center for Type Culture Collection, the preservation address is Wuhan University, Wuhan, China, the preservation date is August 29, 2024, and the preservation number is CCTCC NO: V202476. The strain has high virus titer and can be used for production and inspection of pigeon adenovirus vaccines, and the produced vaccines have good safety and efficacy for prevention of pigeon type 1 adenovirus infection.

The present invention relates to a nanobody directed against the human adenovirus type 4 Hexon protein and its use in antiviral therapy. The nanobody has a unique CDR region, can bind to the adenovirus type 4 Hexon protein with high affinity, and can neutralize adenovirus type 4 pseudovirus with high neutralizing activity in virus neutralization assays. The present invention also provides the use of the nanobody in the preparation of a drug for treating and / or preventing diseases caused by human adenovirus type 4, as well as a diagnostic kit.

The invention relates to a dual LAMP (loop-mediated isothermal amplification) detection system as well as a kit and a detection method thereof. The dual LAMP detection system comprises a primer group, wherein the primer group comprises four primers for specifically detecting the cap gene of the duck circovirus and four primers for specifically detecting the Fiber2 gene of the duck adenovirus type 3. The invention provides a duck circovirus and duck adenovirus type 3 dual LAMP detection system which is efficient, accurate and easy to operate and a corresponding kit. The kit does not need to be operated by complex equipment or professionals, can detect the duck circovirus and the duck adenovirus type 3 at the same time, can introduce a visual detection technology, is suitable for grassroots veterinary laboratory and on-site rapid detection, provides a new detection method for the duck circovirus and the duck adenovirus type 3 for grassroots veterinarians and farmers, and can be used for detecting the duck circovirus and the duck adenovirus type 3 at the same time. Therefore, the requirement on rapid diagnosis of the two viruses in practical application is met.

The application discloses a method for adenovirus whole genome sequencing, a primer group and a kit thereof, and belongs to the technical field of DNA detection.The method for adenovirus whole genome sequencing comprises the following steps: S1, first round amplification is carried out by using first round amplification primers with sequences shown in SEQ ID NO.1 to SEQ ID NO.93 in Table 1; and S2, second round amplification is carried out by using second round amplification primers with sequences shown in SEQ ID NO.94 to SEQ ID NO.285 in Table 2.The average length of data sequenced by the method is 0.8 kb, compared with short read length of a second-generation platform, the uniformity of adenovirus coverage can be greatly improved, meanwhile, the adenovirus types are comprehensively covered, and the method can be widely promoted.

This invention belongs to the technical field of virus detection, specifically relating to a PCR detection kit for the simultaneous detection of four DNA viruses in waterfowl and its application. The PCR detection primer set for the simultaneous detection of four DNA viruses in waterfowl of this invention is the optimal primer set obtained through experimental screening. The kit composed of these primers can rapidly and specifically amplify and diagnose Muscovy duck parvovirus, duck plague virus, duck adenovirus type 3, and duck circovirus simultaneously, exhibiting high specificity and sensitivity. It can accurately and rapidly identify MDPV, DPV, DAdV-3, and DuCV, and can complete quantitative real-time PCR detection in approximately 50 minutes. The kit is simple and practical to operate, achieving efficient simultaneous detection of four viruses, shortening the detection cycle, increasing sensitivity by 10-1000 times, significantly improving early disease diagnosis capabilities, significantly reducing the misdiagnosis rate of diseases in poultry farms, and contributing to the sustainable development of the poultry farming industry.

The present invention belongs to the field of bioengineering technology. To reduce the probability of homologous recombination between a recombinant adenovirus and host cells, a recombinant adenovirus type 5 vector and a method for constructing the vector are provided, which reduces the frequency of reverse mutations. By knocking out the E1B region gene of human adenovirus type 5, and knocking out the gene at base 4090, the length of the homologous sequence between adenovirus type 5 and HEK293 genomic DNA is shortened to 254 base pairs. The deleted E1 region gene fragment includes the pIX-encoding sequence Ad5 3551-4070. The pIX-encoding sequence Ad5 3551-4070 is then reversely inserted into the E4 region, i.e., Ad5 35771. This reduces the frequency of homologous recombination during packaging and amplification of the recombinant adenovirus, thereby reducing the frequency of RCA. This method not only prevents reverse homologous recombination between the recombinant adenovirus vector and HEK293 cell genomic DNA, but also ensures the stability of the recombinant adenovirus particle structure.

The invention belongs to the technical field of molecular detection, and particularly relates to an RPA primer, a probe, a kit and a method for rapidly detecting adenovirus type 4. The RPA primer comprises a forward primer and a reverse primer, the sequence of the forward primer is as shown in SEQ ID No.1, the sequence of the reverse primer is as shown in SEQ ID No.2, and the sequence of the RPA-LFD probe is as shown in SEQ ID No.3. According to the method, the primers and the probe are applied, so that the method has the characteristics of good specificity, high sensitivity, simplicity, rapidness, high efficiency and the like, the requirement of rapid detection in a farm can be met, the detection efficiency is improved, and a new technical support is provided for rapid detection of the chicken adenovirus.

The invention relates to the technical field of biological medicines, in particular to a novel oncolytic adenovirus vector carrying membrane type CD40L as well as a preparation method and application thereof. A plasmid skeleton of the recombinant plasmid is type-5 adenovirus, the recombinant plasmid has an E3 region nucleotide sequence with double deletion of a Gp19k fragment and an ADP fragment, and the E3 region nucleotide sequence comprises a nucleotide sequence of an mCD40L coding region. The oncolytic adenovirus vector can effectively prevent wild type CD40L from being sheared into soluble CD40L after expressing the cell surface, reduce the immunosuppression microenvironment state and reduce the systemic side effects; the compound has a good tumor inhibition effect, and has a wide application prospect in preparation of various preparations for treating tumors.

The present invention relates to an anti-human adenovirus type 4 Hexon protein nanobody and an application thereof, the nanobody having a unique CDR region, capable of binding to the adenovirus type 4 Hexon protein with high affinity, and capable of neutralizing adenovirus type 4 pseudovirus with high neutralizing activity in a virus neutralization test. The invention also provides application of the nano antibody in preparation of a medicine for treating and / or preventing human type 4 adenovirus infectious diseases and a diagnostic kit.

The invention relates to a reagent combination for detecting duck adenovirus type 3 based on RPA-CRISPR / Cas12a as well as a kit and application thereof. The reagent combination comprises a primer pair and crRNA, the primer pair is any one of the four pairs of primer pairs, namely, DAdV-3-Fibir 2-RPA-F1 / R1, DAdV-3-Fibir 2-RPA-F1 / R2, DAdV-3-Fibir 2-RPA-F2 / R3 and DAdV-3-Fibir 2-RPA-F2 / R4, and the primer pair is any one of the four pairs of primer pairs, namely, DAdV-3-Fibir 2-RPA-F2 / R1, DAdV-3-Fibir 2-RPA-F2 / R2, DAdV-3-Fibir 2-RPA-F2 / R2, DAdV-3-Fibir 2- The primers and nucleotide sequences corresponding to the crRNA are as shown in SEQ ID NO: 1-7; according to the present invention, the primer pair and the crRNA combined RPA-CRISPR / Cas12a detection system has characteristics of extremely high specificity and extremely high sensitivity, and can accurately detect DAdV-3 on site.

The present disclosure relates to primer and probe sets for pathogen detection of infection in a transplant patient, a kit and use thereof, and belongs to the technical field of molecular biology detection. There are 23 primer and probe sets that can be used to jointly detect 23 kinds of pathogens with a high infection rate and a high lethality rate after transplantation, including an adenovirus type B; and two ends of a corresponding sequence of each probe have correspondingly a fluorophore and a quencher group, respectively. The present disclosure further provides a real-time fluorescence quantitative PCR kit for pathogen detection of infection in a transplant patient, including the primer and probe sets, a pathogen plasmid standard, a fluorescence quantitative PCR reaction solution, and sterile deionized water, which can simultaneously detect 23 pathogens infected by the transplant patient.

A single stranded nucleic acid aptamer able to specifically bind to at least one Adenovirus type, the aptamer comprising or consisting of one of sequences SEQ ID NO:1 to 3, or variants thereof having at least 50% sequence identity. Additionally, a composition comprising the aptamer. Furthermore, use of the aptamer, for detecting, capturing, concentrating and / or quantifying at least one Adenovirus type. Still further, in vitro methods for capturing, detecting and / or quantifying at least one Adenovirus type. Further yet, a kit for detecting, quantifying, capturing and / or concentrating at least one Adenovirus type.

The invention belongs to the technical field of veterinary biotechnology detection, and particularly relates to a monoclonal antibody for resisting duck adenovirus 3 type Fiber2 protein and application of the monoclonal antibody to detection or identification of duck adenovirus 3 type. The duck adenovirus 3 type Fiber2 protein is taken as an immunogen to screen a hybridoma cell strain with good reactivity to different strain whole viruses of the duck adenovirus 3 type, an antibody generated by the hybridoma cell strain is analyzed, the monoclonal antibody resisting the duck adenovirus 3 type Fiber2 protein is obtained, and the sequence information is shown in a table 3 in the specification. The monoclonal antibody provided by the invention can identify different strains of the duck adenovirus 3, does not generate cross reaction with other common poultry disease viruses such as fowl adenovirus, novel duck reovirus and the like, has good specificity and sensitivity, can be used for detecting the exogenous virus of the duck adenovirus 3 in poultry virus live vaccines, and has good application prospects. And the kit can also be used for clinical identification, virus content determination and epidemiological investigation of the duck adenovirus type 3.

The invention belongs to the technical field of virus detection, and particularly relates to a composition for specific detection of a monoclonal antibody for type-5 adenovirus and application of the composition. The antibody combination comprises 8B9 and 4A6 which respectively target different epitopes of HADV-5 Hexon protein, and is suitable for a double-antibody sandwich ELISA (enzyme-linked immuno sorbent assay) detection system. Wherein 8B9 is used as a coating antibody, 4A6 is used as a detection antibody marked by HRP, high-sensitivity and high-specificity detection of HADV-5 and replication-deficient recombinant adenovirus 5 can be realized through cooperation of 8B9 and 4A6, and cross reaction with other common adenovirus types is avoided. The invention also provides a variable region amino acid sequence and a nucleotide sequence of the antibody, and an ELISA detection method based on the antibody combination. The method is simple and convenient to operate, good in repeatability and suitable for detecting the type 5 adenovirus.

The invention provides a probe, a primer, a kit, a method and application for human adenovirus whole genome sequencing. Aiming at a conserved sequence in an adenovirus genome, a plurality of pairs of primer probes are explored. According to the primer pair and the probe, the coverage degree of different adenovirus type genomes is greatly improved, and meanwhile, the amplification capacity of the primer aiming at targets with different lengths is enhanced. Eight adenovirus whole-genome sequences can be uniformly amplified, so that not only is the coverage degree of the whole-genome sequences improved, but also the capturing capacity for targets with different lengths is enhanced. In addition, the method has high tolerance for samples with different qualities, samples with low concentration or samples degraded to a certain degree can be subjected to high-uniformity total-sequence capture, and the method has high sensitivity and uniformity.

The present invention relates to a nanobody directed against the human adenovirus type 4 Hexon protein and its use. The nanobody has a unique CDR region, can bind to the adenovirus type 4 Hexon protein with high affinity, and can neutralize adenovirus type 4 pseudovirus with high neutralizing activity in virus neutralization tests. The present invention also provides the use of the nanobody in the preparation of a drug for treating and / or preventing diseases caused by human adenovirus type 4, as well as a diagnostic kit.

Provided is an oncolytic adenovirus (Ad) of which the basic skeleton is an Ad having a different serotype from that of an adenovirus type-5 (Ad type-5), and which has high virus yield and can exhibit a satisfactory anti-tumor effect. Also provided is a pharmaceutical composition containing the oncolytic Ad as an active ingredient. The oncolytic Ad is a recombinant oncolytic Ad characterized in that a part or the whole of E1 gene of an Ad type-5 is incorporated into an E1 region in the genome of an Ad having a different serotype from that of the Ad type-5 and a part or the whole of E4 gene of the Ad type-5 is incorporated into an E4 region in the genome of the Ad having a different serotype from that of the Ad type-5.

To provide methods for treating human patients with replicable oncolytic adenovirus.SOLUTION: Disclosed is a method comprising the steps of: systemically administering multiple doses of a parenteral formulation of a replicable oncolytic adenovirus of subgroup B in a single treatment cycle, where the total dose given in each dose is in the range of 1×1010-1×1014 viral particles, and where each dose of virus is administered over a period of 1 to 90 minutes, for example, at a rate of viral particle delivery in the range of 2×1010 particles per minute to 2×1012 particles per minute. The disclosure further extends to formulations of the oncolytic adenoviruses and combination therapies of the viruses and formulations with other therapeutic agents.SELECTED DRAWING: None

The invention relates to the technical field of biological detection, in particular to a primer and probe combination for detecting and identifying tortoise adenovirus type 1-3, and the primer and probe combination comprises an upstream primer, a downstream primer and a probe for detecting or / and identifying the tortoise adenovirus type 1-3; the nucleotide sequences of the upstream primers of the tortoise serrated adenovirus type 1-3 are respectively as shown in SEQ ID NO: 1-3, the nucleotide sequences of the downstream primers of the tortoise serrated adenovirus type 1-3 are respectively as shown in SEQ ID NO: 4-6, and the nucleotide sequences of the probes of the tortoise serrated adenovirus type 1-3 are respectively as shown in SEQ ID NO: 7-9. The primer probe combination disclosed by the invention can be used for real-time fluorescent quantitative PCR and multiple PCR technologies, the type-1-3 of the serrate turtle adenovirus can be simultaneously detected in a one-tube PCR reaction system, and three typing detection results of the serrate turtle adenovirus can be obtained by only carrying out one-time nucleic acid extraction and one-time PCR detection on one sample. The method is simple, accurate in result and easy to popularize. Meanwhile, the primer and the probe disclosed by the invention are high in sensitivity and good in specificity.

The invention discloses a method for efficiently and stably producing a duck adenovirus type 3 vaccine antigen and application of the duck adenovirus type 3 vaccine antigen. The method comprises the following steps: resuscitating LMH cells, inoculating the resuscitated LMH cells into a DMEM / F-12 liquid culture medium, and carrying out shake culture to obtain an LMH cell suspension; inoculating a duck adenovirus type 3 seed virus into the LMH cell suspension, and carrying out shake culture to obtain a virus suspension; and inoculating the virus suspension into a suspension reactor containing an LMH cell suspension, and carrying out enlarged suspension culture to obtain the duck adenovirus type 3 vaccine antigen. The method for producing the duck adenovirus 3 type vaccine antigen provided by the invention is simple and convenient to operate and stable in quality, the production efficiency of the DAdV-3 antigen can be remarkably improved, and the production cost can be saved; the method for producing the duck adenovirus type 3 vaccine antigen can be applied to preparation of inactivated vaccines, is good in safety and high in immunogenicity, and is suitable for large-scale production of DAdV-3 vaccines in workshops.

The present invention relates to a nanobody directed against the human adenovirus type 4 Hexon protein and its use. The nanobody has a unique CDR region, can bind to the adenovirus type 4 Hexon protein with high affinity, and can neutralize adenovirus type 4 pseudovirus with high neutralizing activity in virus neutralization tests. The present invention also provides the use of the nanobody in the preparation of a drug for treating and / or preventing diseases caused by human adenovirus type 4, as well as a diagnostic kit.

The invention discloses a recombinant serum type 3 duck adenovirus and a construction method and application thereof, the recombinant serum type 3 duck adenovirus replaces a non-structural protein gene ORF52 in the serum type 3 duck adenovirus with an exogenous target gene or inserts the exogenous target gene into the non-structural protein gene ORF52, and the nucleotide sequence of the non-structural protein gene ORF52 is shown as SEQ ID NO. 1. The recombinant serum type 3 duck adenovirus for expressing green fluorescent protein is constructed by targeting an exogenous gene EGFP to a non-structural protein gene ORF52 of the duck adenovirus type 3 by utilizing a gene editing technology, and the non-structural protein gene ORF52 is proved to be capable of serving as an insertion site of the exogenous gene, so that the recombinant serum type 3 duck adenovirus can be used as an insertion site of the exogenous gene. And a foundation is laid for developing a genetic engineering recombinant DAdV-3 multi-combined multivalent vaccine based on taking DAdV-3 as a virus vector.

The present invention discloses a test strip for detecting neutralizing antibodies to human adenovirus type 5. The test strip comprises a PVC base plate, and a sample pad, a fluorescent pad, a chromatographic membrane, and an absorbent pad arranged in sequence above the PVC base plate. The chromatographic membrane comprises a solid-phase nitrocellulose membrane consisting of a test line and a quality control line. The test line is coated with mouse anti-human IgG antibodies, the quality control line is coated with goat anti-rabbit IgG antibodies, and the fluorescent pad is coated with quantum dot-labeled rabbit IgG and quantum dot-labeled antigens. The quantum dot-labeled antigens are either or both of quantum dot-labeled human adenovirus type 5 fiber knob protein and quantum dot-labeled human adenovirus type 5 intact inactivated virus. The present invention also discloses a method for preparing the test strip, as well as a test card and kit using the same. The present invention can be used to rapidly assess the level of neutralizing antibodies to adenovirus type 5 in a population, providing guidance for adenovirus vector therapy methods, and the like.

The invention provides an adenovirus vector as well as a construction method and application thereof, and the recombinant adenovirus vector is named as pAd5-C7. According to the invention, a Tet gene regulation and control system and a Cumate gene regulation and control system are integrated into a single-copy plasmid pAd5 containing an adenovirus type 5 genome in different ways to obtain a recombinant adenovirus vector pAd5-C7. On the basis of a Rembineering technology, a screening gene SacB / beta-geo is inserted into the downstream of a CMV promoter on pAd5-C7, and a rapid, efficient and universal recombinant adenovirus vector construction platform is constructed. Recombinant adenovirus plasmids constructed on the basis of the platform can be used for obtaining recombinant adenovirus particles with advanced protein regulated and controlled. In an adenovirus vector based on the human adenovirus type 5, the recombinant adenovirus which is normally expressed in allowable cells and is inhibited in advanced protein expression of the adenovirus in non-allowable cells is prepared.