172 results about "Mycoplasma hyorhinis" patented technology

The invention provides a human mycoplasma pneumoniae gold-marked silver-stained immunochromatographic assay kit and a preparation method and application thereof. The assay kit comprises a detection card and a silver-stained sensitivity-enhanced pad, wherein the detection card is composed of a bottom plate, a sample pad, an absorbent pad, a conjugate pad and a detection layer; the conjugate pad is coated with a colloidal gold-marked polyclonal antibody mixture of colloidal gold marked rabbit anti-human mycoplasma pneumoniae P1 protein and P30 protein; the detection layer is composed of a solid phase nitrocellulose membrane with a detection line and a quality control line; the detection layer is bonded on the bottom plate, the conjugate pad and the absorbent pad are partially overlapped with the detection layer respectively and are bonded with the detection layer and the bottom plate respectively; the sample pad and the conjugate pad are partially overlapped to be bonded with the conjugate pad and the bottom plate respectively; and the silver-stained sensitivity-enhanced pad consists of a AgNO3 pad and a restoring pad. The human mycoplasma pneumoniae gold-marked silver-stained immunochromatographic assay kit can effectively improve the detection sensitivity of the human mycoplasma pneumoniae, has the strong specificity and has the high application value in the aspects of clinical diagnosis of human mycoplasma pneumoniae, etiology identification, epidemiological investigation and the like.

The present invention provides a mycoplasma vaccine, its preparation and application thereof. The foregoing mycoplasma vaccine comprises inactivated Mycoplasma hyorhinis ATIT-7 only or the mixture of inactivated Mycoplasma hyorhinis ATIT-7 and inactivated Mycoplasma hyopneumoniae, which effectively prevents the infection of swine enzootic pneumonia in pigs.

The invention discloses a goat TLR4 gene knock-out vector and a construction method thereof. The construction method comprises the following steps: firstly, designing an sgRNA fragment of the TLR4 gene by adopting a CRISPR / cas9 system, synthesizing an sgRNA nucleotide sequence, constructing and simultaneously expressing the sgRNA and plasmid PYSY-sgRNA of Cas9 D10A, connecting and transforming to an Escherichia coli DH5 alpha competent cell, and verifying the transformant; and judging and proving by enzyme digestion and sequencing that the TLR4 gene knock-out vector is constructed correctly. The invention adopts the CRISPR / cas9 for constructing the vector, and provides a theoretical basis for subsequently acquiring a goat TLR4 gene deletion type alveolar epithelial cell system, and studying the immune response molecular mechanism of mycoplasma pneumonia infection.

The invention provides a mycoplasma hyopneumoniae culture medium and a preparation method thereof, belonging to the technical field of veterinary biology. The mycoplasma hyopneumoniae liquid culture medium comprises the components as follows: brain heart infusion, lactalbumin hydrolysate, PPLO (pleuropneumonia-like organism) broth, yeast extract powder, proteose peptone, sodium thiosulfate, Hank's liquid, sodium pyruvate, 0.1% phenol red solution, penicillin and deionized water. The preparation method comprises the following steps of: adding health horse serum before using, and adding agar into the liquid culture medium to obtain a solid culture medium of mycoplasma hyopneumoniae. The viable bacteria titer of the mycoplasma hyopneumoniae culture medium can reach 1*109CCU / ml-1*1010CCU / ml; the viable bacteria titer and the separation sensibility are far higher than those of the existing culture medium, and the mycoplasma hyopneumoniae is fast in growth speed and high in the separation sensibility; and the preparation method of the culture medium is simple in technology, strong in operability, and suitable for industrial large-scale production.

The present invention provides a one phase, aqueous vaccine composition for immunizing an animal against infection by Mycoplasma hyopneumoniae, comprising: an immunizing amount of a Mycoplasma hyopneumoniae bacterin, an acrylic acid polymer in the concentration range between 0.8 and 1.2 mg / ml, and a pharmaceutically acceptable carrier, and substantially no oil. It is especially useful for immunizing a pig against infection by Mycoplasma hyopneumoniae for at least 20 weeks after a single administration, which effective immunity is reached within 4 weeks after vaccination.

The invention provides a kit for jointly detecting respiratory tract pathogen through a multiple fluorescent PCR method. The kit comprises six components: reaction liquid A, reaction liquid B, reaction liquid C, enzyme mixed liquid, positive control and negative control, and comprises 11 common respiratory tract pathogen detections (general type of influenza virus A, influenza virus B, respiratory syncytial virus, 1 / 2 / 3 type of human parainfluenza virus, adenovirus, mycoplasma pneumoniae, chlamydia pneumonia, legionella pneumophila, streptococcus pneumonia, haemophilus influenza, A streptococcal); the amplification is performed through three reaction buffers, and each reaction buffer contains four fluorescent channels, 90% pathogen infection on the clinic can be checked.

The invention discloses a mycoplasma bovis monoclonal antibody, and a preparation method and an application thereof. The preparation method of the mycoplasma bovis monoclonal antibody comprises the following steps: 1, preparing a mycoplasma bovis antigen; 2, immunizing mice by the antigen; 3, preparing a hybridomas cell secreting the mycoplasma bovis monoclonal antibody and monoclonal antibody ascites; and 4, purifying the above obtained monoclonal antibody. In the invention, mice are immunized by a mycoplasma bovis geographical strain HB0801 antigen, a hybridomas cell strain 1C11, CCTCC NO:C201218, which can efficiently secrete the mycoplasma bovis monoclonal antibody, is obtained through a hybridomas cell technology, the generated monoclonal antibody can be specifically combined with mycoplasma bovis and mycoplasma agalactiae, and the monoclonal antibody is utilized to establish sandwich ELISA for detecting the mycoplasma bovis antigen. The method has the advantages of simple operation, short required time and high sensitivity.

The invention relates to a primer composite for detecting respiratory pathogens. The primer composite comprises at least one group of a mycoplasma pneumoniae group, a chlamydia pneumoniae primer group, an influenza A / B virus primer group, a parainfluenza virus primer group, an adenovirus primer group and a respiratory syncytial virus primer group. The invention further relates to a kit comprising the primer composite. The kit further comprises a micro-fluidic chip wrapping primers, and a macroscopic indicator. The invention further relates to a detection method adopting the primer composite. The detection method comprises the steps of primer composite coating, to-be-detected sample nucleic acid extraction, LAMP reaction and visual result interpretation. The kit and the method are applied to the micro-fluidic chip for visual judgment, instant detection of the seven respiratory pathogens is rapidly and accurately realized, and a result is judged by a naked eye by color change, so that the kit and the method are simpler, more convenient and quicker in practical applications, are easy to operate, and are suitable for site operation.

The invention provides a combined inactivated vaccine against mycoplasma hyopneumoniae (MHP) and mycoplasma hyorhinis. The combined inactivated vaccine contains the MHP and the mycoplasma hyorhinis with preferred content as well as a Carbomer adjuvant with concentration of 10% (V / V). The invention further provides a preparation method of the combined inactivated vaccine against the MHP and the mycoplasma hyorhinis. The preparation method comprises the following steps: preparing production strains; preparing bacterium solution for producing seedlings; and inactivating, concentrating and blending to obtain the vaccine. The combined inactivated vaccine has the advantages of strong specificity and good immunity, thus solving the problem of specific infection caused by the MHP in the current domestic breeding farm and obtaining the mycoplasma hyorhinis vaccine under a blank state at home and abroad at present. The combined inactivated vaccine has the beneficial effects that the step of vaccine inoculation is simplified, trouble caused by a plurality of inoculation and easily produced side effects are avoided, and vaccine cost is saved, thus being especially applicable to preventing andtreating mixed infection diseases in the breeding farms at home and abroad and the like; and compared with the existing single vaccine, application range is widened and immune effect is enhanced.

The invention relates to a preparation method of a mycoplasma gallisepticum and mycoplasma synoviae bivalent inactivated vaccine. The method comprises the steps as follows: a mycoplasma gallisepticum virulent CR strain and a mycoplasma synoviae HN01 strain which have good immunogenicity are inoculated on a proper culture medium for cultivation, so that a culture is acquired; and the culture is inactivated through a formaldehyde solution and then is mixed with an oil emulsion adjuvant and emulsified, so that the mycoplasma gallisepticum and mycoplasma synoviae bivalent inactivated vaccine is prepared. The mycoplasma gallisepticum and mycoplasma synoviae bivalent inactivated vaccine is used for preventing mycoplasma gallisepticum and mycoplasma synoviae diseases, and can realize immunization, prevent two pathogens at the same time and reduce the workload of immunization; and the prepared vaccine is stable in performance, good in immune effect, and more suitable for actual production of China.

This invention provides an immunogenic composition including a soluble portion of a Mycoplasma hyopneumoniae (M.hyo) whole cell preparation, wherein the soluble portion of the M.hyo preparation is substantially free of both (i) IgG and (ii) immunocomplexes comprised of antigen bound to immunoglobulin.

The invention discloses a colloidal gold method detection test strip and a reagent kit for an IgG antibody of a respiratory disease and a preparation method of the reagent kit. The test strip determines the IgG antibody by using a principle of an immunocapture method; respiratory herpes viruses, adenoviruses, influenzaviruses and an IgG antibody of mycoplasma pneumoniae can be detected jointly by one operation; the operation process is simplified; the test strip is simple, convenient, rapid and accurate in detection, suitable for mass detection and applicable to primary screening and epidemiological survey; and a result is distinct and easy to distinguish.

The present invention relates to a Mycoplasma hyopneumoniae vaccine strain comprising a mutation in at least one of the genes listed or as deposited with the National Measurements Institute (Australia) under accession number NM04 / 41259, which strain is temperature sensitive and attenuated, a vaccine comprising such strains and methods and uses thereof.

The invention provides a vaccine composition, which comprises an immune amount of a porcine circovirus antigen, an immune amount of a mycoplasma hyopneumoniae antigen, and levamisole or its derivative, and a polymer. The vaccine composition not only can have an immunological enhancement effect on two antigens at the same time, but also has stability, and can be used for dilution of porcine reproductive and respiratory syndrome live vaccines.

Belonging to the molecular biology field, the invention relates to a primer set for respiratory tract infection pathogen detection, a rapid diagnostic kit and a detection method. The primer set can achieve one-time detection of the following 9 respiratory tract infection pathogens: influenza A virus, influenza B virus, respiratory syncytial virus, parainfluenza virus type I, parainfluenza virus type II and parainfluenza virus type III, adenovirus, mycoplasma pneumonia, legionella pneumonia, chlamydia pneumonia and human rhinovirus. The invention adopts solid phase recombinase-polymerase constant temperature gene amplification method to detect respiratory multiple infection pathogens. The primer set adopted by the invention for respiratory tract infection pathogen detection has strong specificity and high amplification efficiency, can effectively and rapidly detect the 9 pathogens simultaneously, and realizes multiplex detection.

The invention belongs to the technical field of zoonosis prevention and treatment, and particularly relates to a mycoplasma bovis mutant strain with a growth defect under cell coculture and application. The mycoplasma bovis Mbov-0328 gene deletion mutant strain T9.386 is sifted out from a mycoplasma bovis gene mutant library, and the gene is coded to form cyclic dinucleotide phosphodiesterase. When the mutant strain and a bovine embryo pneumonocyte are co-cultured, the remarkable growth defect phenotype is shown; the mutant strain shows small colonial morphology on a PPLO solid culture medium.Proteomics between the mutant strain and a wild strain shows a remarkable difference expression spectrum. The mutant strain has 38 differential expression proteins, wherein 30 proteins are in up-regulated expression, and 8 proteins are in down-regulated expression. The mutant strain can be applied to the field of mycoplasma bovis metabolic physiology, pathopoiesia and immune prevention.

The invention relates to a detection method for specific antibodies IgM of mycoplasma pneumonia and influenza viruses based on a micro-fluidic chip and application of the detection method. The method is characterized by taking the micro-fluidic chip with a plurality of pipelines as a reaction platform and detecting by using a chemical luminescent signal so as to independently or simultaneously detect the serum specific antibodies IgM of mycoplasma pneumonia and influenza viruses. As the micro-fluidic chip is combined with chemiluminiscence, the detection method integrates the advantages of rapidness, high efficiency and small sample amount of the micro-fluidic chip and high specificity of chemiluminiscence, also has the advantages of simplicity in operation, low cost, rapidness and the like, is a multi-target, real-time and high-sensitivity detection method and can be applied to rapid diagnosis of pneumonia and influenza.

The invention discloses an indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) diagnostic kit of mycoplasma gallisepticum, which comprises an ELISA plate coated by species specificity proteins PvpA, an elution buffer solution, an antibody diluting solution, an ELISA secondary antibody, a substrate color developing solution and a stop solution. The kit disclosed by the invention selects the species specificity proteins PvpA, covers the high-frequency variation regions: DR-1 and DR-2 regions of the PvpA proteins, has high specificity and immunogenicity, improves the specificity and sensitivity of the detection result, lowers the production cost and is suitable for popularization and use in grassroots veterinarian mechanisms.

The invention discloses a mycoplasma bovis immunity-related protein P22, a nucleotide sequence for coding the same and an application thereof. A mycoplasma bovis Hubei strain is taken as a sample, and an efficient two-dimensional electrophoresis system of a mycoplasma bovis whole protein is established, so that a two-dimensional electrophoretogram with high resolution and high repeatability is obtained; an immunity-related protein is screened in combination with Western blot, and is named P22 protein; and as proved by a prokaryotic expression result of the protein, a Western blot rest result of a recombinant protein and a mycoplasma bovis positive serum is negative, and a Dot blot test result is positive. The protein is proved to be possibly a conformation-dependent immunity-related protein of mycoplasma bovis, and the mycoplasma bovis immunity-related protein P22 plays a guidance role in diagnosing, preventing and treating a mycoplasma bovis disease.

The invention discloses a LAMP (loop-mediated isothermal amplification) kit for detection of mycoplasma pneumoniae and a special LAMP primer for detection of mycoplasma pneumoniae. The special LAMP primer for detection of mycoplasma pneumoniae is designed according to a specificity conservative target sequence of a mycoplasma pneumoniae P1 gene (GenBank number: CP002077.1). The LAMP primer is formed by six primers including outer primers MP-16F3 and MP-16B3, inner primers MP-16FIP and MP-16BIP and loop primers MP-16LF and MP-16LB. By the aid of the LAMP kit and the special LAMP primer for detection of mycoplasma pneumoniae, quickness, convenience, high efficiency, high specificity and high sensitivity in qualitative detection of the mycoplasma pneumoniae in samples of pure bacteria, sputum, bronchoalveolar lavage fluid, throat swabs and the like can be realized without complicated instruments, and a new technical platform is provided for detection of the mycoplasma pneumoniae.

The invention provides a mycoplasma pneumonia mosaic antigen amino acid sequence containing an amino acid sequence as shown in SEQ ID NO:1 as well as a full-gene synthesized mycoplasma pneumonia mosaic antigen full-gene sequence containing an amino acid sequence as shown in SEQ ID NO:2. The invention also provides a method for constructing the two gene sequences. The invention also provides a preparation method of the mycoplasma pneumonia mosaic antigen containing full-gene synthesis, a mycoplasma pneumonia detection kit and a preparation method thereof. Mp recombinant mosaic antigen is selected as a mark material and is applied to a gold immunochromatography system, and the detection system is directly marked and captured, so the sensitivity is greatly improved, the specificity of the antigen is high, the antigen is easy in cultivation and purification, and cost is reduced; and a new method for detecting mycoplasma pneumoniae IgG rapidly and accurately is provided for clinical use, and a good market prospect is achieved.

The invention provides a primer for detecting sheep mycoplasma pneumonia. The sequence of the primer comprises an upstream primer: 5'-GGGACTTCGGGACTTATTGGA-3', and a downstream primer: 5'-CACGAGATGCAAACTGATTTACTTG-3'. According to a p80 gene sequence of KM435069.1 logged in GenBank, a specific primer is designed, a real-time fluorescence quantification PCR (polymerase chain reaction) method is established for a p80 gene of sheep mycoplasma pneumonia for a first time, and the method is good in specificity, high in sensitivity and good in repeatability and can be applied to diagnosis and epidemiological investigation on sheep mycoplasma pneumonia.

The invention relates to a Chinese medicinal composition for treating ovine mycoplasma pneumoniae, which comprises the following components in part by weight: 15 to 25 parts of Vietnamese sophora root, 15 to 25 parts of indigowoad root, 15 to 25 parts of indigowoad leaf, 5 to 15 parts of baical skullcap root, 15 to 20 parts of barbed skullcap herb, 5 to 15 parts of caladium, 15 to 20 parts of Indian buead, 20 to 30 parts of astragalus, 15 to 20 parts of szechuan tangshen root, 10 to 20 parts of Szechuan lovage rhizome and 5 to 15 parts of liquoric root. In the Chinese medicinal composition, a pure Chinese medicinal herb preparation is developed by combining the research achievements of the pharmacology of modern Chinese veterinary medicines, formulating prescriptions elaborately and screening strictly according to the treatment based on syndrome differentiation and holism concept in Chinese veterinary science. The Chinese medicinal composition consists of the following 11 Chinese medicinal herbs of the Vietnamese sophora root, the indigowoad root, the baical skullcap root, the barbed skullcap herb, the caladium, the Indian buead, the astragalus, the Chinese angelica, the szechuan tangshen root, the Szechuan lovage rhizome, the liquoric root and the like, and has the effects of clearing heat, detoxicating, relieving cough, reducing sputum, tonifying qi and promoting blood circulation.

The invention relates to a kit for detecting mycoplasma hominis nucleic acid through a PCR-fluorescence probe method and a detecting method of the kit. The kit comprises a nucleic acid extracting set and a nucleic acid amplification detecting set. The nucleic acid amplification detecting set comprises PCR reaction liquid, negative control and positive control. The PCR reaction liquid comprises DNA polymerase, UNG enzymes, reaction buffer liquid, a primer, a probe and Mg2+. The positive control comprises positive plasmids and human genomes. The negative control comprises human genomes. The probe is a Taqman fluorescence probe. The special primer probe is designed according to an MH polymerase conservative target sequence by means of the kit and the method, the MH is rapidly detected through the PCR-fluorescence probe method, and the result is more reliable and accurate. Compared with an existing culture method, operation is easy and convenient and the speed is fast when mycoplasma hominis is identified; the whole process is completed within 3 hours; specificity and flexibility are high, and the result is visual.

The invention discloses a lyophilized reagent for nucleic acid diagnosis. The lyophilized reagent comprises a lyophilized protecting agent which contains 2%-10% of trehalose, 2%-5% of glucan, 0.5%-2%of PEG8000 and 0.01mg-0.02mg / ml of BSA protein, and the lyophilized protecting agent contains mycoplasma pneumoniae, chlamydia pneumoniae and adenovirus primers and probes. The lyophilized reagent hasadvantages that since the lyophilized reagent contains three pathogen nucleic acids of mycoplasma pneumoniae, chlamydia pneumoniae and adenovirus, amplification detection of various pathogen nucleicacids can be realized, namely three targets can be simultaneously detected by one reaction system, and accordingly reagent and detection consumables are greatly saved, high detection throughput is realized, and a clinical application range is expanded. In application of the lyophilized reagent for lyophilization treatment, liquid nitrogen is adopted for quick pre-freezing of a liquid reagent, so that a subsequent lyophilization process is shortened, and stability of the lyophilized reagent is improved.

The invention discloses a primer sequence combination and a method for applying the primer sequence combination in PCR amplification. The amplification target in the method is specific fragments EMA inside the genome of Elizabethkingia meningoseptica, and the method shows excellent specificity and sensibility; and moreover, by using the method, the neisseria meningitidis, haemophilus influenzae, staphylococcus aureus, streptococcus pneumoniae, escherichia coli, listeria monocytogenes, mycoplasma pneumoniae, bordetella pertussis, klebsiella pneumoniae and mycobacterium tuberculosis and the other pathogenic bacteria which are likely to cause meningitis and diseases of the upper respiratory tract, and easy to be confused with the Elizabethkingia meningoseptica in an identification of bacteria, can be distinguished at one time. Additionally, the lower detection limit of the method can reach 10-4 ng per Mul.