62 results about "Mycoplasma synoviae" patented technology

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.

The invention discloses a bi-fluorescence quantitative RT-PCR detection kit for mycoplasma synoviae and avian reovirus. The kit comprises a primer group and a probe group, wherein the primer group comprises a primer 1, a primer 2, a primer 3 and a primer 4, which respectively have base sequences of sequence tables SEQ.ID.No.1 and 2, and SEQ.ID.No.4 and 5; and the probe group comprises a probe A and a probe B which respectively have the base sequences of sequence tables SEQ.ID.No.3 and 6. Experiments prove that the kit disclosed by the invention has the advantages of short detection time, high detection sensibility and strong specificity, two pathogens of the mycoplasma synoviae and the avian reovirus can be detected and discriminated at the same time, and the corresponding pathogeny content can be quantified, so that the kit can be used for assessing the curative effects of vaccines and medicines on the mycoplasma synoviae and the avian reovirus and researching pathogenic mechanisms and other respects; therefore, the kit disclosed by the invention has an important significance on prevention and control of the mycoplasma synoviae and the avian reovirus.

An H95 attenuated mycoplasma synoviae strain is obtained through in-vitro subculture in order to research an attenuated vaccine capable of preventing mycoplasma synoviae. The H95 strain and a white oil adjuvant are mixed in the proportion of 1:1 to prepare the attenuated vaccine, and the immune effect of the strain on chickens is researched by immunizing the chickens with different immune doses; animal infection research is carried out through a pathogenicity wild strain, so that the prevention effect of the attenuated vaccine prepared from the H95 attenuated strain on mycoplasma synoviae is verified. The invention provides a preparation method of the attenuated vaccine capable of preventing mycoplasma synoviae and an effective immunizing dose. A scientific basis is provided for clinicallypreventing mycoplasma synoviae.

The invention discloses a multi-fluorescent immunoassay method for rapidly distinguishing 6 types of poultry respiratory pathogens. The multi-fluorescent immunoassay method is simple to operate; a target amplified fragment is obtained through a PCR (Polymerase Chain Reaction); then an amplified product, fluorescence coded microspheres and streptavidin-phycoerythrin are hybridized; an MFI (Mean Fluorescence Intensity) value is read through a detector to distinguish viruses of different types. According to the method disclosed by the invention, avian influenza viruses, chicken infectious bronchitis viruses, chicken Newcastle disease viruses, chicken infectious laryngotracheitis viruses, mycoplasma gallisepticum and mycoplasma synoviae can be accurately detected at the same time; the multi-fluorescent immunoassay method has high specificity, high sensitivity and good repeatability. Compared with a traditional detection method, the method disclosed by the invention realizes simultaneous detection of a plurality of types of different target molecules in the same sample; the use amount of the sample is less; the method is simple and rapid to operate and the detection cost can be greatly reduced.

The invention discloses a multiple fluorescence immunoassay method for detecting mycoplasma gallisepticum and mycoplasma synoviae. The multiple fluorescence immunoassay method has the advantages that the operation is simple; a target amplified fragment is obtained through PCR (polymerase chain reaction), then the amplified fragment, fluorescent encoding microspheres, and streptavidin-phycoerythrin are hybridized, and an MFI (mean fluorescence intensity) value is read through a detection instrument, so as to distinguish different types of pathogens; the mycoplasma gallisepticum and mycoplasma synoviae can be simultaneously detected and identified; the specificity is strong, the sensitivity is high, the repeatability is good, and the like; multiple types of molecules with different purposes in the same sample can be simultaneously detected; the flexibility is good, and the type of to-be-detected pathogens can be increased or decreased on the basis according to requirements.

The invention discloses a product, method and application for simultaneously detecting mycoplasma gallisepticum and mycoplasma synoviae. According to the product and the method, a common specific protein antigen of mycoplasma gallisepticum and mycoplasma synoviae is preferably an outer membrane protein WP-041352022.1 as a detection object, a PCR primer pair S1 is designed and screened on the genome DNA level of the protein, and the PCR primer pair has the advantages of strong specificity, high sensitivity and no primer dimer generation. The invention provides a product and a method for simultaneously detecting two pathogens by using one pair of PCR primers, mutual interference between multiple PCR primers and the product is avoided, the detection rate is improved, and the method has the advantages of economy, simplicity, convenience and high efficiency.

The invention discloses a multiple fluoroimmunoassay method capable of quickly differentiating infectious laryngotracheitis virus (ILTV), infectious bronchitis virus (IBV), myeoplasma gallisepticum (MG) and mycoplasma synoviae (MS) and a reagent. The method is simple to operate, a target amplified fragment is obtained through polymerase chain reaction (PCR); an amplified product, fluorescence coded microspheres and streptavidin-phycoerythrin are hybridized, a mean fluorescence intensity (MFI) value is read through a detector, and pathogens of different types are differentiated. According to the method, avian infectious laryngotracheitis virus, infectious bronchitis virus, myeoplasma gallisepticum and mycoplasma synoviae can be accurately detected simultaneously, and the method is high in specificity and sensitivity and good in repeatability. Compared with a conventional detection method, the method can be used to simultaneously detect various target molecules in an identical sample, the sample usage amount is small, the method is simple and quick to operate, and the detection cost can be greatly reduced. The method is good in flexibility, and the varieties of detected pathogens can be increased or decreased as required on the basis.

The application of the invention provides an immune composition and a subunit vaccine. The immune composition contains chicken mycoplasma synoviae MSPA (Mycoplasma synoviae surface protein A) proteincoded with a nucleic acid molecule of which the nucleotide sequence is as shown in SEQID NO:1 or a nucleic acid molecule of which the nucleotide sequence is 95% or above the same as that as shown in SEQID NO:1, and mycoplasma synoviae MSPB (Mycoplasma synoviae surface protein B) protein coded with a nucleic acid molecule of which the nucleotide sequence is as shown in SEQID NO:3 or a nucleic acidmolecule of which the nucleotide sequence is 95% or above the same as that as shown in SEQID NO:3. Sf9 cells of the vaccine are used for expressing MSPB protein and MSPA protein, the antigenicity, theimmunogenicity and the function of the product namely the immune composition are similar to those of natural protein, and the immune composition is high in expression level and high in immunogenicity, does not have pathogenicity to chickens; and besides, the vaccine can be prepared through large-scale serum-free suspension culture with a bioreactor, and besides, the production cost of the vaccinecan be greatly reduced.

The invention discloses an LAMP (loop-mediated isothermal amplification) visual detection kit for chicken mycoplasma synoviae (MS). The invention provides an LAMP primer group for detecting the chicken mycoplasma synoviae. The LAMP primer group comprises a primer 1, a primer 2, a primer 3 and a primer 4, wherein the nucleotide sequences of the primer 1, the primer 2, the primer 3 and the primer 4 are respectively a sequence 1, a sequence 2, a sequence 3 and a sequence 4 in a sequence table. Experiments in the invention proof that the primer provided by the invention, the prepared MS-LAMP detection reagent and the created method have the characteristics of speediness, flexibility, specificity and simpleness, only one temperature controllable water bath kettle is used, and results can be determined by eyes instead of an instrument, so that the LAMP visual detection kit disclosed by the invention is applicable to rapid detection in a food products factory, a border port, a farm and a primary-level veterinary station, is significant in effective prevention and control of MS and has better application prospect.

The present invention provides a Mycoplasma synoviae indirect ELISA detection kit. The kit uses main membrane antigen MSPB recombinant protein amplified by a strain of Mycoplasma synoviae Ningxia isolate through overlapping PCR as a coating antigen. The kit is capable of rapidly, specifically and efficiently detecting mycoplasma synoviae, to monitor prevalence of Mycoplasma synoviae in chicken flocks.

The invention discloses a RPA kit for rapidly detecting mycoplasma synoviae of chicken and its use. The kit comprises: a reaction tube containing RPA lyophilized particles, a reaction buffer solution,magnesium acetate, a pair of primers and a probe. The experiments prove that the sensitivity and specificity of the kit of the present invention are comparable to those of a qPCR method, and the entire reaction time is completed within 20 minutes, and the detection result only needs to be determined by the change of a fluorescence curve. Therefore, the kit of the invention has the advantages of quickness, high efficiency, sensitivity, and the like, provides a technical means for effectively detecting and identifying the mycoplasma synoviae of chicken, is easy and fast to operate, and is suitable for clinical veterinary on-site diagnosis and scientific research on mycoplasma synoviae of chicken.

The invention relates to a mycoplasma synoviae culture medium and a preparation method thereof. The culture medium comprises a basal culture medium and auxiliary components. The basal culture medium comprises a phosphate buffer solution, glucose, lactoalbumin hydrolysate, coenzyme I, arginine hydrochloride, L-cysteine hydrochloride, MEM, yeast extract powder, 1% phenol red, and trehalose. The auxiliary components comprise pig serum and penicillin. According to the culture medium, the antigen content of mycoplasma synoviae bacterial liquid is remarkably increased, the bacterial liquid titer reaches up to 1012.0-13.0 CCU/ml. Meanwhile, the separation rate of the MS wild strains is remarkably improved and is as high as 93%. According to the preparation method of the culture medium, the antigen content of mycoplasma synoviae is increased, the production process is simplified, and the production cost is reduced.

The invention discloses a salmonella pullorum and mycoplasma synoviae double-plate agglutination antigen as well as a preparation method and application thereof, and the preparation method comprises the following steps: respectively carrying out resuscitation passage on salmonella pullorum C79-1 and mycoplasma synoviae GX11-T to prepare seed bacterial liquid, inoculating the seed bacterial liquid into a corresponding culture medium, carrying out enlarged culture, harvesting, respectively inactivating by a formaldehyde solution, concentrating, mixing the two bacterial liquids according to a certain volume ratio, adding a crystal violet solution with the final concentration of 3% by mass fraction for dyeing and glycerol with the volume fraction of 10%, uniformly mixing, and subpackaging, thereby obtaining the product. The technological method is simple and reasonable, low in cost and good in stability, the prepared double-plate agglutination antigen can detect two epidemic disease antibodies at the same time through one-time reaction, the advantages of being high in sensitivity, rapid in diagnosis, high in specificity, easy to observe the agglutination effect and the like are achieved, the detection time and the detection cost are remarkably saved, and the detection efficiency is improved. The product can be clinically used for rapidly detecting the two antibodies of the pullorum disease and the mycoplasma synoviae, diseased chickens are eliminated, and disease purification is achieved.

The invention discloses a chicken mycoplasma synoviae inactivated vaccine which comprises an antigen and a vaccine adjuvant, and the antigen is an inactivated mycoplasma synoviae MS-FJ01 strain; the strain is separated from sick chicken attached joints with chicken mycoplasma synoviae in Fuzhou, and belongs to a typical strain in Fujian province; the strain is preserved in the China Center for Type Culture Collection, the preservation address is Wuhan, China, the preservation number is CCTCC NO: M 2021210, and the preservation date is March 8, 2021; according to the scheme, the vaccine can generate a high-level antibody, the immunization duration is long, the morbidity of the immunized chicken mycoplasma synoviae is obviously reduced, and the prevalence of the chicken mycoplasma synoviae can be effectively prevented; the chicken mycoplasma synoviae inactivated vaccine prepared from the mycoplasma synoviae MS-FJ01 strain in the scheme can effectively prevent and control the chicken mycoplasma synoviae, the richness of commercially available vaccines of the category is improved, and an optional scheme is provided.

The present disclosure relates to a composition for preventing and treating Mycoplasma synoviae infection, the composition comprising Tuf, NOX, MS53-0285 or a combination thereof as active ingredients. The composition of the present disclosure is effective in alleviating symptoms caused by Mycoplasma synoviae infection and can be used as an effective tool in preventing or treating poultry diseases.

The invention discloses a preparation method of a mycoplasma synoviae monoclonal antibody. The conservative analysis is carried out on two imaginary membrane proteins with the mycoplasma membrane surface protein sizes of 20Kda and 13Kda; then recombinant plasmids synthesizing, protein inducible expression, and affinity chromatography purification are carried out; the purified recombinant protein is used as immunogen to immunize BALB/c mice, cell fusion is carried out after antibody detection is qualified, a single monoclonal antibody cell strain is obtained after subcloning of fusion cells, and the prepared monoclonal antibody can well and specifically recognize mycoplasma synoviae antigen.

The invention provides a mycoplasma synoviae antigen protein LP85, an established ELISA method for detecting a corresponding antibody of mycoplasma synoviae, an ELISA kit prepared according to the method and a corresponding using method. Based on the mycoplasma synoviae antigen protein LP85, especially a mycoplasma synoviae recombinant expression protein LP85, the ELISA detection method of the corresponding antibody and the ELISA kit prepared according to the method have the characteristics of better specificity and sensitivity and good detection repeatability compared with an existing detection form.

The invention provides a mycoplasma synoviae virulent strain and application thereof. A chicken flock infected by a certain synovial sac in Fujian Province is subjected to MS separation, purification and identification and then named as an MS FZ strain, SPF chickens infected by a pure culture of the MS FZ strain and aged 10-11 weeks have high pathogenicity, and at least 80% of the SPF chickens have air sacculitis; and the MS FZ strain is inactivated through formaldehyde and then emulsified with an oil adjuvant to prepare an inactivated vaccine, the inactivated vaccine is used for immunizing the SPF chickens twice, then a fresh culture of the MS FZ strain is used for counteracting toxic substances to verify the vaccine protection force, and the result shows that at least 80% of the SPF chickens obtain immune protection. Therefore, the separated MS FZ virulent strain can be used as a virulent strain for inspection of effectiveness evaluation of different drugs or prevention and treatment methods and can be used as a vaccine candidate strain for preparing an MS inactivated vaccine.

The invention belongs to the technical field of PCR, and particularly relates to a PCR detection kit for rapidly identifying mycoplasma hyopeumoniae, mycoplasma synoviae and mycoplasma hyorhinis, and applications of the PCR detection kit. The PCR detection kit comprises PCR premix buffering solution, superpure water, Marker DL2000, upstream primer, downstream primer, a negative control and a positive control. The sequence of the upstream primer is 5'-ACACCATGGGAGCTGGTAAT-3'; the sequence of the downstream primer is 5'-GTTCATCGACTTTCAGACCCAAGGCAT-3'. The PCR kit has the advantages of being low in detection limit, good in sensitivity, strong in specificity and the like, can rapidly detect and identify mycoplasma hyopeumoniae, mycoplasma synoviae and mycoplasma hyorhinis.

The invention relates to a mycoplasma synoviae culture medium and a preparation method thereof, and belongs to the technical field of veterinary biology. The mycoplasma synoviae culture medium is composed of a basic culture medium and an auxiliary culture medium, the basic culture medium mainly comprises Hank's concentrated solution, milk protein hydrolysate, yeast extract, beef heart extract powder and trehalose, and a high-pressure steam sterilization mode is used for sterilization; the auxiliary culture medium is prepared from arginine, sodium pyruvate, coenzyme I, penicillin and porcine serum, and all the components of the auxiliary culture medium are filtered and sterilized. The titer of the mycoplasma synoviae cultured by the culture medium disclosed by the invention reaches 1010-1011CCU/ml, the culture time is 14-24h, the content of porcine serum is as low as 5%, and the stress reaction of heterologous to chickens and the production cost of enterprises are greatly reduced. The inactivated vaccine prepared from the mycoplasma synoviae cultured by the culture medium has good safety and immunogenicity.

The invention relates to the field of animal medicines, and particularly discloses a plant extract composition for inhibiting mycoplasma synoviae, and a preparation method and a use method of the plant extract composition. The plant extract composition for inhibiting the mycoplasma synoviae of the chicken comprises plant extracts, and the plant extracts comprise one or a combination of multiple of a dry aloe extract, a semen raphani extract, a folium cortex eucommiae extract, a radix scutellariae extract, a fructus forsythiae extract, a Chinese pulsatilla root extract, a rhizoma ligustici extract and a cortex acanthopanacis extract. The plant extract composition for inhibiting the mycoplasma synoviae has the advantage of effectively inhibiting the mycoplasma synoviae, so that the mycoplasma synoviae is prevented and controlled, and the morbidity of the mycoplasma synoviae is reduced. Meanwhile, the plant extract composition for inhibiting the mycoplasma synoviae is not easy to generate drug resistance, and has the advantages of being natural, free of toxic and side effects, free of drug resistance and free of residues.

The invention discloses an enrofloxacin nano-microemulsion solution for treating Mycoplasma synoviae disease in poultry. The nano-microemulsion is prepared by the following method: preparing raw materials including 5-15 parts by weight of enrofloxacin, 5-15 parts by weight of an oil phase, 5-20 parts by weight of a surfactant, 5-20 parts by weight of a cosurfactant, and 30-80 parts by weight of awater phase; adding enrofloxacin, the surfactant and the cosurfactant into a reaction device, and dispersing in a water phase through stirring, ultrasonic treatment or homogenization treatment to obtain an emulsion; slowly adding the emulsion into an oil phase, introducing nitrogen to expel oxygen, and adding a mixture of initiators; and carrying out external cooling and polymerization on the above reaction to obtain the enrofloxacin nano-microemulsion solution. The invention aims to provide an enrofloxacin nano-microemulsion solution for treating Mycoplasma synoviae disease in poultry. The process effect of adopting a nano-microemulsion preparation is more stable than the effect of a common oral solution, and the enrofloxacin nano-microemulsion solution shows a better therapeutic effect on Mycoplasma synoviae disease in poultry.

The invention relates to an avian mycoplasma culture medium, a preparation method of an avian mycoplasma solution and applications of the avian mycoplasma culture medium and the avian mycoplasma solution, and belongs to the technical field of biology. The avian mycoplasma culture medium comprises a basal culture medium and auxiliary components, wherein the basal culture medium is a conventional avian mycoplasma liquid culture medium, and the auxiliary components comprise gelatin, Tween 80 and calcium chloride; a strain for production is mycoplasma galliscepticum or mycoplasma synoviae, the avian mycoplasma solution is produced by solution inoculation, culture and harvesting, and an avian mycoplasma vaccine is prepared. According to the preparation method, antigen content of the avian mycoplasma solution is increased, production technology is simplified, production cost is reduced, probability of occurrence of vaccine immunity side effects is reduced, and vaccine immunity effect is improved.