92 results about "BCG vaccine" patented technology

The invention belongs to the biomedical detection field, in particular relates to a reagent and method used for detecting activity tuberculosis and latent tuberculosis infection; based on the genomic principle, the invention discloses a novel detection reagent for mycobacterium tuberculosis, containing protein or polypeptide which is represented by SEQ ID1-2, 4-5, 8-28; the method uses one or a plurality of SEQ ID 1-28 protein or polypeptide to contact T cells of a mycobacterium tuberculosis host, and detects cytokine released from the T cells; the method can detect the tuberculosis and latent infection effectively and is not interfered by BCG vaccine at the same time; the invention also discloses a diagnostic reagent kit and other application based on the protein or polypeptide and the method; compared with the T-SPOT of the prior art, the invention can improve detectable rate obviously under the condition that the specificity is not reduced; the reagent kit has cheap price, and the cost is 1/5 to 1/10 of that of the T-SPOT reagent, thus being beneficial to being popularized in the developing countries and poor areas.

The recombinant protein vaccine is a fusion protein formed from connection of BCG vaccine heat shock protein 65 with 1-5 copies of human prostatic specific antigen cytotoxin T lymphocyte polyepitope,in which the single-copy polypeptide of said human prostatic specific antigen cytotoxin T lymphocyte polyepitope possesses amino acid sequence showed by SEQ ID NO:2, and the double single-copy polypeptide of the human prostatic specific antigen dcytotoxin T lymphocyte polyeptope possesses amino acid sequence showed by SEQ ID NO:4. After it is applied in human body, it can effectively prevent and cure carcinoma of prostate. Said invention also provides the gene for coding said two kinds of recombinant protein vaccines.

The present invention relates to molecular immunology technology, and aims at screening out antigen epitope molecular simulation peptide capable of exciting human body's protective immunoreaction against tubercle bacillus, researching protective immunoreaction mechanism against tuberculosis, and further developing new type of concatenate polyepitope tuberculosis vaccine. The present invention provides one kind of antigen epitope molecular simulation peptide capable of exciting human body's protective immunoreaction against tubercle bacillus, and the peptide contains the amino acid sequence selected from SEQ ID Nos. 2, 5, 10, 12, 14 and 15. The present invention also provides the screening process and use of the peptide. The present invention may be used in preventing and controlling tuberculosis.

The invention discloses a reagent for detecting tubercle bacillus infection in vitro and a method thereof. The reagent comprises M233 polypeptide represented by SEQ ID No.1; and cytokine released from T cells is detected by contacting the M233 polypeptide or an analog thereof with the T cells of a tubercle bacillus host to determine whether the T cells identify the M233 polypeptide or the analog thereof. The reagent has the advantages of high sensitivity, good specificity, being free from interference of BCG vaccine and non-tuberculosis mycobacteria vaccine, being capable of detecting active pulmonary tuberculosis patients, the patients with dormant infection and healthy persons who contact with mycobacterium nontuberculosis. The reagent and the method are especially applicable to detecting tuberculosis and/or dormant infection thereof for Chinese people.

The invention belongs to the technical field of animal infectious disease gene engineering and discloses a bovine tuberculosis antibody detecting immune colloidal gold test strip prepared by utilizing RV3872, ESAT6 and CFP10 fusion protein, and a preparation method and application. A colloidal gold immunochromatographic test strip is established by using the fusion protein as a colloidal gold labeled antigen and a capture antigen in a detection region of a nitrocellulose membrane. The detection of the bovine tuberculosis antibody by using test strip has prominent advantages of strong specificity and high sensitivity, and simultaneously bacillus calmette-guerin immunity and nontuberculosis mycobacteria infection can be identified and detected. The test strip comprises recombinant Escherichia coli BL21/pET28a-MPBrce, and the strain expresses mycobacterium bovis RCE proteins and is preserved in the China Center for Type Culture Collection with the collection number of CCTCC No:M208244.

The invention provides polypeptides encoded by open reading frames present in the genome of Mycobacterium tuberculosis but absent from the genome of BCG and diagnostic and prophylactic methodologies using these polypeptides.

A recombinant protein vaccine which is a fusion protein of BCG vaccine's heat shock protein 65 and the core antigen of multi-epitope hepatitis-C virus, its amino acid sequence and nucleotide sequencefor coding it, the expression carrier containing said nucleotide sequence, the host cell containing said expression carrier, the preparing process of said recombinant protein vaccine, the vaccine containing said recombinant protein for preventing and treating hepatitis C, and a method for detecting the activity of specifically killing T, lymphocytes by the hepatitis C induced by said vaccine and its cell model are disclosed.

The present invention discloses a recombinant protein vaccine which is a fusion protein prepared from the BCG vaccine heat-shock protein 65 and multi-epitope HER-2 antigen and can be used for effectively preventing and treating breast cancer. The gene for coding the said recombinant protein vaccine is also disclosed.

Disclosed is a pharmaceutical composition for treating tuberculosis which is prepared from the following Chinese medicinal herbs (by weight ratio): Radix Ranunculi Ternati 1-3 weight parts, toad venoms 1-3 weight parts, rhubarb horsetails 1-3 weight parts, fritillary 1-3 weight parts, selfheal 1-3 weight parts, asparagus root 1-3 weight parts. The invention also discloses the process for preparing the pharmaceutical composition.

A vaccine for treating or preventing the establishment of latent tuberculosis infections is provided. The vaccine comprises a recombinant mycobacterium that overexpresses the transcription factor DosR, at a level sufficient to induce production of the dosR regulon genes or proteins. A host to whom the vaccine is administered mounts an immune response to the dosR regulon proteins and is thus protected from the establishment, persistence or reactivation of latent tuberculosis.

The invention discloses a mycobacterium tuberculosis fusion protein and an application thereof in induction of peripheral blood mononuclear cells (PBMCs) to generate cytokines. The fusion protein includes three proteins PPE41, ESAT-6 and PE25, and the proteins are connected through connecting peptides. Compared with present stimulants, the fusion protein provided by the invention has the advantages of efficient effect, strong sensitivity, high specificity and good stimulation effect. The fusion protein stimulates the PBMCs to generate a large amount of mycobacterium tuberculosis antigen specific IFN-gamma, IL-2, TNF-alpha and other tuberculosis related factors, and the above stimulation induction reaction is free from BCG vaccine interference. The fusion protein can effectively improve the tuberculosis detection rate and is of positive significance to control the tuberculosis. The fusion protein can be applied in researches of the tuberculosis pathopoiesis and immunoprophylaxis mechanisms and control of the tuberculosis as a stimulant.

The invention provides a recombination BCG vaccine rBCG::AB for protecting against tubercle bacillus and a preparation method thereof. The preparation method comprises the following steps: firstly, amplifying full-length genes containing promoters and signal peptides of Ag85A and Ag85B proteins, and establishing a recombination colibacillus-mycobacteria shuttle expression plasmid after subcloning; and then, transforming the plasmid in a BCG vaccine to form the recombination BCG vaccine rBCG::AB which can simultaneously excessively express the Ag85A and Ag85B proteins. The recombination BCG vaccine rBCG::AB realizes the high-abundance overexpression of the Ag85A and Ag85B proteins in the culture filtrate of the recombination BCG vaccine. After the immunization of animals, the recombination BCG vaccine rBCG::AB can obviously induce cellular immune response to the Ag85A and Ag85B proteins and provide stable and persistent protection against infection, thereby overcoming the defects of short protection time, instable protection against adult tuberculosis and the like of the traditional BCG vaccine.

The invention discloses a recombinant interferon--BCG Vaccine bacterial strain and its preparing method, transferring the composed pshuttle phIFN-alpha-2B into the BCG to compose Rbcg-hIFN-alpha-2B bacterial strain. Because it can secrete the interferon IFN-alpha-2B, it can reduce the usage of BCG as compared with that of the wild BCG on the condition of reaching the same and even increased immune effect, thus reducing toxic and side effect; and it can secrete cell factors at the properest time and part. Therefore, it lays the foundation of application to curing cystic tumors.

The invention provides construction of TRAIL (Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand) recombinant bacille calmette guerin (rBCG), and relates to a shuttle expression vector comprising a signal peptide fragment of a major secretory antigen Ag85B of BCG and a gene fragment of a TRAIL and a construction method thereof. The obtained shuttle expression vector pMV261-Ag85B-TRAIL is used for constructing rBCGTRAIL, and can be applied to preparation of TRAIL rBCG for treating superficial bladder tumors, preventing postoperative recurrence thereof and preventing tuberculosis. The rBCG has dual functions of TRAIL and BCG, so that cooperative and synergistic actions of the TRAIL and BCG can be better brought into play; rBCG-TRAIL can secrete TRAIL, and the using amount of the rBCG-TRAIL can be lower than that of the BCG under the condition that the same or better immune effect is achieved, so that the toxic or side effect is reduced; and the rBCG-TRAIL can directly secrete TRAIL efficiently on a certain part, so that tumor cells can be killed in cooperation with the rBCG-TRAIL, and high cost caused by the use of a foreign cell factor is avoided.

The invention relates to an Escherichia coli-mycobacterium shuttle vector plasmid suitable for secretory expression of pathogenic microorganism antigen and application thereof to preparation of a pathogenic microorganism vaccine. The Escherichia coli-mycobacterium shuttle vector plasmid is constructed based on an Escherichia coli-mycobacterium shuttle vector plasmid pBCG-2000; a modified mycobacterium tuberculosis heat shock protein 60 promoter regulation sequence with SEQ ID NO.1 as the sequence is inserted; a T4 transcription termination sequence which is artificially synthesized and has the sequence of SEQ ID NO.2 is inserted; a modified signal peptide sequence, with SEQ ID NO.3 as the sequence, of a BCG antigen gene 85b is inserted; and an artificial synthetic sequence with SEQ ID NO.4 as the sequence is added. The Escherichia coli-mycobacterium shuttle vector plasmid can be applied to the research of recombinant vaccines of resisting Toxophasma gondii, SARS coronaviruses and tubercle bacillus.

The invention discloses a two-stage type microneedle array patch capable of simultaneously realizing BCG vaccine inoculation and diagnosis. The two-stage type microneedle array patch comprises a microneedle array and a patch; the microneedle array is arranged on the patch; the microneedle comprises a soluble drug-loaded microneedle tip and an insoluble microneedle supporting end; the soluble drug-loaded microneedle tip is prepared by mixing a BCG vaccine, tubercle bacillus element microspheres and soluble auxiliary materials and performing freeze drying; and the tubercle bacillus element microspheres are prepared by combining slow-release auxiliary materials and tubercle bacillus elements. The BCG vaccine and the tubercle bacillus elements are gathered at the soluble microneedle drug-loaded tip at the same time; and the release time of the tubercle bacillus elements is controlled through a slow-release microsphere technology, so that two drug administration purposes of inoculating theBCG vaccine and diagnosing whether the BCG vaccine is successfully inoculated or not can be achieved through one-time drug administration. The invention further discloses a preparation method of the patch.

Methods and compositions for the detection and diagnosis of infectious diseases are provided. In particular, efficient and sensitive methods and compositions for the detection of active mycobacterial disease are provided for distinguishing between individuals having active disease, and individuals who have been immunologically exposed, such as those infected with a mycobacterium but are without active disease, or those who have been vaccinated with BCG. The methods comprise topical application of antigen compositions for transdermal delivery.

The invention discloses an sRNA marker for distinguishing mycobacterium tuberculosis and a BCG vaccine and a use thereof. For solving the problem that mycobacterium tuberculosis infection and BCG vaccination are difficult to distinguish, the type of sRNA in mycobacterium tuberculosis is detected and screened by bacterial RNA sequencing firstly, a specific primer and kit capable of detecting the sRNA of mycobacterium tuberculosis are designed according to the screened target sRNA sequence, and a corresponding real-time fluorescence quantitative PCR detection method is established. After free sRNA is extracted from bacterial bodies and cDNA is synthesized, the content of sRNA is detected by the real-time fluorescent quantitative PCR method. The experiment proves that the method can be used for distinguishing the mycobacterium tuberculosis and the BCG vaccine, has the advantages of high sensitivity, good specificity, low cost, convenience in operation and the like, and is hopeful to further meet the clinical and laboratory needs for rapid diagnosis of the mycobacterium tuberculosis and the BCG vaccine.

The invention provides rBCG for expression of Br. Melitensis P39 and L7/L12 fusion gene. The rBCG is constructed by transferring an expression vector carrying codon-optimized Br. Melitensis P39 and L7/L12 fusion gene into BCG. Brucellosis-generated cytoplasm binding protein PBP39 (coding gene is P39) and Brucellosis ribosomal protein L7/L12 are both T-cell antigen. Bacillus Calmette-Guerin (BCG) vaccine is the only one commercial vaccine for preventing tuberculosis so far. The BCG vaccine has a remarkable immunologic adjuvant effect and is an exogenous gene expression host with good performance and high safety. By BCG expression of the codon-optimized Brucellosis P39 and L7/L12 fusion gene, expression quantity of the target gene can be increased. The rBCG vaccine can simulate intracellur infection and parasitic characteristics of Brucellosis to more effectively induce body to generate immune response, can perform advantages of high safety, simple preparation, low cost, etc. of BCG as the expression host as well as the immunologic adjuvant effect of the BCG itself, and is expected to become a novel Brucellosis vaccine.

The invention relates to a Chinese medicine compound prescription for treating chronic hepatitis B and a method for preparing the same. The Chinese medicine compound prescription consists of mile swertia, serissa foetida comm, astragalus root and radix salivae miltiorrhizae; the materials are decocted by water and extract to obtain a compound active extract. The Chinese medicine compound prescription has the protecting function on liver injury, can reduce the level of ALT and MDA of serum in liver, raise the activity of SOD of serum and has effect of treating hepatitis caused by bcg vaccine. The Chinese medicine compound prescription can be used for preparing a drug with the function of treating chronic hepatitis B.

The invention discloses bcg vaccine dry powder microneedle vaccine and a preparation method thereof. A microneedle consists of bcg vaccine dry powder, a hollow microneedle array and a substrate patch; the bcg vaccine dry powder is loaded in the hollow microneedle array. The method comprises the following preparation steps that (1) the bcg vaccine is cultured in a 7H9 liquid culture medium; when a logarithmic phase is reached, bacteria are harvested through centrifugation; freeze drying is performed; the materials are ground into powder; (2) sodium hyaluronate salts with the mass concentration being 15 percent is placed into a microneedle mold; after the horizontal centrifugal drying, a hollow microneedle head is prepared; then, the bcg vaccine dry powder is filled in the microneedle; the sodium hyaluronate salts with the mass concentration being 10 percent is added at the surface of the mold; centrifugation, drying and demolding are performed. The bcg vaccine dry powder can be stored in the microneedle for 90 days at normal temperature; the loading quantity is great; the immune effect is good; adverse reaction such as generation of red and swollen skin, bubble, scar, inflammation and the like when the bcg vaccine is injected can be avoided; the transportation and storage cost is greatly reduced; during inoculation, the operation by professionals is not needed; no biological waste materials are generated.

The present invention discloses a human Mycobacterium tuberculosis fusion protein and applications thereof. The fusion protein concurrently contains ESAT6/CFP10/TB7.7 polypeptide sequences, such that the name is the fusion protein ect. The fusion protein can stimulate human T cells to release gamma-interferon (IFN-gamma). With the application of the fusion protein ect, the active tuberculosis and the latent tuberculosis infected person can be detected, and the people being subjected to BCG vaccine (BCG) vaccination can be excluded.