43 results about "Mycoplasma capricolum" patented technology

The invention aims at providing a primer and a probe for detecting peste des petits ruminants virus by virtue of a RPA (Recombinase Polymerase Amplification) technique. According to the primer and the probe, a forward primer sequence is represented by SEQ ID NO:1, a reverse primer sequence is represented by SEQ ID NO:2, and a probe sequence is represented by SEQ ID NO:3. According to a method, a large number of primers and probes are designed according to genomic sequences of the peste des petits ruminants virus, and a pair of primer and probe combinations capable of rapidly detecting nucleic acid of the peste des petits ruminants virus are screened out; obvious amplification curves can be obtained by carrying out rapid detection by virtue of the primer and the probe and taking nucleic acid RNA of the peste des petits ruminants virus of domestically separated Tibet strains and Xinjiang strains as a template, and no amplification curve is obtained through a reaction by taking other pathogenic nucleic acids such as foot and mouth disease virus, contagious pustular dermatitis virus, mycoplasma capricolum and bluetongue disease virus as the template.

The invention discloses a rapid detection kit for goat contagious pleuropneumonia and a preparation method thereof. The kit is established on the basis of molecular biology. The kit comprises 2*loop-mediated isothermal amplification (LAMP) reaction liquid, a primer mixer, BstDNA polymerase, a color-developing agent, a standard positive template and ddH2O, wherein the primer mixture comprises a forward inner primer FIP, a downward inner primer BIP, a forward outer primer F3 and a downward outer primer B3; and the ratio of the FIP to the BIP to the F3 to the B3 is 8:8:1:1. The kit has the characteristics of high sensitivity and specificity and the like, can react quickly, is easy to operate, does not need special instrument equipment and overcomes the defects of long pathogeny separation time, high workload, fussy detection method, complicated operation and the like of goat contagious pleuropneumonia, and the detection method is complicated, is difficult to operate and the like, and can be used for detecting whether clinical samples are infected with Mycoplasma capricolum subsp. Capripneumonia as well as performing early diagnosis and molecular epidemiological survey of goat contagious pleuropneumonia.

The present invention relates to contagious caprine plueropneumonia (CCPP) antibody detection test strips. The test strips comprise: a PVC liner, a nitrocellulose membrane, an absorption pad, a gold standard pad and a sample pad. The left end of the nitrocellulose membrane is sprayed with anti-mycoplasma capricolum subsp. capripneumoniae mccp IgG, wherein the IgG is adopted as a quality control line. The right end of the nitrocellulose membrane is sprayed with mycoplasma capricolum subsp. capripneumoniae mccp polysaccharide antigen, wherein the polysaccharide antigen is adopted as a detection line. The gold standard pad is sprayed with colloidal gold-labeled mycoplasma capricolum subsp. capripneumoniae mccp polysaccharide antigen. The preparation process comprises: a, preparing the colloidal gold, wherein a sodium citrate reduction method is adopted to prepare the colloidal gold; b, preparing the colloidal gold-labeled mycoplasma capricolum subsp. capripneumoniae mccp polysaccharide antigen; c, assembling the antibody detection test strip. According to the present invention, the presences of the detection line and the quality control line are adopted to determine whether the CCPP antibody exists; the test strips of the present invention have characteristics of convenient detection, fastness and accuracy, and are applicable for primary level quarantine, epidemiological investigation quarantine, isolated quarantine and rapid detection so as to eliminate infected sheep in time, control disease occurrence, and reduce economic losses.

The invention provides a real-time fluorescent quantitative PCR detection primer used for mycoplasma capricolum subsp.capripneumonia. The primer sequences are as follows: the sequence of a forward primer is 5'-GAACTGAAGAAGGTATGGCTGAAGA-3', and the sequence of a reverse primer is 5'-CCTGTTCCAGCACCAACTAAAAC-3'. According to the arcD gene sequence of AY529462.1 logged in the GenBank, the specific primer is designed, a SYBR Green I real-time fluorescence quantitative PCR method of the mycoplasma capricolum subsp.capripneumonia is firstly built at home and abroad, and the method is good in specificity, high in sensitivity and good in repeatability.

The invention discloses a bivalent inactivated vaccine for goat mycoplasma pneumonia and chlamydia psittaci and a preparation method thereof. The active ingredients of the bivalent inactivated vaccinecomprise inactivated Mycoplasma capricolum subsp. Capripneumoniae, inactivated mycoplasma ovipneumoniae and inactivated MOMP protein of chlamydia psittaci. The amino acid sequence of the MOMP proteinof the chlamydia psittaci is shown as a sequence 2 in a sequence table. The mycoplasma ovipneumoniae is mycoplasma ovipneumoniae SH-01 CGMCC No.16503. The Mycoplasma capricolum subsp. Capripneumoniaeis Mycoplasma capricolum subsp. Capripneumoniae GT-01 CGMCC No.16504. Experiments prove that the bivalent inactivated vaccine has good safety in immunizing experimental animals and target animals, and can effectively prevent goat mycoplasma pneumonia and chlamydia psittaci from infecting the experimental animals and the target animals. The vaccine has important significance for preventing and controlling prevalence and propagation of mycoplasma pneumonia and chlamydia psittaci of goats. The vaccine has important application value.

The invention which belongs to the technical field of biological detection relates to a method for detecting MP of sheep and provides the methylation state of CpG islands in an MBL (mannan-binding lectin) gene promoter region related with the MP of sheep and an application thereof. The invention also provides a method for detecting the MP of sheep and concretely relates to the methylation detection of the MBL promoter region and the artificial infection verification. The methylation state detection of the MBL gene promoter region of sheep is carried out through selecting a bisulfite sequencing process, the methylation state and the frequency of each CpG locus in a given region can be accurately detected, three substantially differential CpG methylation loci, CpG3, CpG4 and CpG5, related with the MBL level are found, a case that the MBL level after toxic elimination is lower than the MBL level before the toxic elimination is promoted, and the MBL gene promoter region methylation may be a more substantial characteristic of the MBL level reduction of serum. The method allows the MP infection of sheep and susceptible individuals to be sensitively and accurately detected, and provides complete information and scientific bases for the sheep MBL gene promoter region methylation and the MP infection research of sheep.

The invention discloses a mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium. The mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium comprises the following components of PPLO broth, sodium pyruvate, lactose, tryptone, fresh yeast extract, L-glutamine, L-cysteine, lactoalbumin hydrolysate, horse serum, penicillin, phenol red, and deionized water; and the invention also provides a preparation method. The mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium has the beneficial effect that the serum content of the mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium is only 5%, which is 1 / 5-1 / 4 of the serum amount of the mediums in the prior art; in addition, the prepared semi-finished product bacteria liquid titer can reach 1010 CCU / ml, which is obviously higher than that of the medium in the prior art. The low-serum high efficiency medium used for culturing the mycoplasma capricolum subsp. Capripneumoniae is in favor of mitigating allergic stress reaction of heterogenous serum on goat, increases the biological safety, increases the live bacteria titer, and reduces the production cost.

The invention provides a kit for rapid detection and genotyping of mycoplasma pneumoniae, belonging to the technical field of microbiological detection. The kit contains 2 specific primer pairs and 2 probes matched with the primer pairs, and the nucleotide sequences are shown by SEQ ID No.1-6 respectively; rapid detection and genotyping of mycoplasma pneumoniae are realized by a duplex fluorescent quantitative PCR (polymerase chain reaction) method, the detection sensitivity is 3CFU, the coincident rate between the genotyping accuracy and that of common genotyping methods reaches 100%, the genotyping sensitivity is much higher than that of common culture-dependent genotyping technologies, the genotyping time is greatly shortened, and the genotyping can be performed at the time of detection on mycoplasma pneumonia in a sample; and moreover, the shortcomings of low sensitivity, time and labor consuming and relatively high cost of the existing mycoplasma pneumoniae genotyping method are overcome, and the kit provided by the invention has good practical sample detection and genotyping ability and is suitable for popularization and application.

The invention discloses a high-flux micro-fluidic chip for detecting pathogeny of the epidemic disease of the respiratory tract of pigs and a detection method and belongs to the technical field of biological chips. The high-flux micro-fluidic chip comprises an LAMP amplification reagent, a DNA / cDNA template, ddH2O, LAMP positive control, LAMP negative control, a detection chip, a chip probe and amicro-fluidic chip analyzer, the detection chip is placed in the constant-temperature amplification micro-fluidic chip analyzer, and the chip probe comprises nucleotide sequences shown in SEQ ID NO:1-42 and specific nucleic acid segments applied to detection of the hog cholera virus, porcine reproductive and respiratory syndrome virus, porcine circovirus 2 type, porcine circovirus 3 type, haemophilus parasuis, mycoplasma hyopneumoniae, porcine pasteurella and swine influenza virus. According to the high-flux micro-fluidic chip, detection can be completed within one hour, the detection limit is5*10-2ng / microliter, different kinds of pathogeny of the epidemic disease are accurately distinguished, and the high-flux micro-fluidic chip has the advantages of quick detection, high sensitivity and accurate specificity.

The invention discloses a culture medium for separation and purification of mycoplasma bovirhinis. Every 1000ml of culture medium comprises, by weight 10.5g of PPLO (pleuropneumonia-like organism) broth, 8,0g of cerebrocardiac leach liquor, 4.0g of sodium chloride, 0.05g of magnesium sulfate, 0.2g of potassium chloride, 0,09g of calcium chloride, 0.024g of disodium hydrogen phosphate, 0,03g of monopotassium phosphate, 5.0g of glucose, 5.0g of lactoalbumin hydrolysate, 60ml of 25% of fresh yeast leach liquor, 100ml of aseptic swine serum, 100ml of aseptic horse serum, 100mg of ampicillin and 1ml of 10% of thallium acetate. The invention further provides a preparation method and application of the culture medium. Compared with the prior art, the culture medium for separation and purification of the mycoplasma bovirhinis, the preparation method and the application of the culture medium have the advantages of high growing speed and titer and high mycoplasma bovirhinis separation success rate.

The invention relates to a synthesize prevention and control method of enzootic pneumonia of pig farms and belongs to the technical field of disease prevention and control. The synthesize prevention and control method includes: inoculating mycoplasma hyopneumoniae vaccine for each pig at 7-21 days old; adding houttuynia cordata and florfenicol into feed when the pigs are at 60-70 days old, and feeding the pigs for ten continuous days; adding houttuynia cordata and florfenicol into feed when the pigs are at 100-110 days old, and feeding the pigs for ten continuous days; at the whole breeding stage of the piglets, controlling the temperature of the piggery at 18-28 degrees centigrade and the humidity at 55-65%. The synthesize prevention and control method of enzootic pneumonia of pig farms set up comprehensive prevention and control measures of the enzootic pneumonia at all the breeding stages of the pigs, accordingly the enzootic pneumonia of the pig farms is effectively prevented and controlled, and the morbidity of enzootic pneumonia is reduced by more than 65%.

The invention discloses a method for simply comparison of the virulence of goat mycoplasma goat pneumonia subspecies strains. The method comprises the steps: inoculating the resurgent goat mycoplasmagoat pneumonia subspecies strains of equal concentration to chicken embryos, then putting the inoculated chicken embryos in a thermostat box for cultivation, detecting the status and death situationsof the chicken embryos periodically, counting the fatality of the chicken embryos inoculated with the strains, and adopting the fatality for comparison and judgment. The method has the advantages of simple operation, low cost, no need for complicated conditions of animal breeding, easy availability of experimental animals, a short consumed time, good repeatability and the like, and the workload ofanimal experiments is reduced greatly, so that the defects of high price of experimental animal goats, large individual differences, high breeding requirements, complex experimental operation, a longexperiment cycle and high workload in the prior art are overcome.

The invention discloses a poultry and waterfowl one-day-age hatching device purifying spray. The poultry and waterfowl one-day-age hatching device purifying spray is composed of tiamulin, neomycin sulfate, nuramyl dipeptide and taurine. The poultry and waterfowl one-day-age hatching device purifying spray has the advantage that the poultry and waterfowl one-day-age hatching device purifying sprayis applied in a sprayed mode inside a hatching device, and after being fully atomized, achieves drug delivery via lung absorption to purify one day old baby chicks or ducklings, thereby combining breeding environment, chicken purifying situation and the like in China, taking into full consideration of pharmacokinetic factors of drugs as well as nutrition demands of the baby chicks, achieving comprehensive purifying effects on both bacteria prone to vertical propagation among chicks such as mycoplasmas and salmonellas, and bacteria prone to infection during hatching such as staphylococcus aureus, pseudomonas aeruginosa and Escherichia coli, taking into account lack of energy of chicks during transportation as well as injuries of respiratory tract villi and the like caused by stress of transportation, comprehensively improving the health of chicks, reducing incidence of diseases and enhancing food safety.

The invention relates to application of a Chinese medicinal composition in preparing a gel for treating gynecological inflammation, wherein, the Chinese medicinal composition is prepared by the following bulk medicines by weight: 2000 parts of cortex ailanthi, 1500 parts of sophora flavescens, 1500 parts of frankincense, 50 parts of borneol, and 1500 parts of cortex moutan. The gel prepared by the Chinese medicinal composition in the invention can overcome many defects of chemical medicines, exert the incomparable extensive curative effects (such as heat clearing and dehumidifying, leucorrhea stopping and itching relieving, necrotic tissue removing and granulation promoting) of chemical medicines, and can be widely applied in treating gynecological inflammation (such as damp-heat pouring downward leukorrhagia including bacterial vaginosis, vulvovaginal candidiasis, etc.) caused by bacteria, fungi, protozoa, chlamydiae, mycoplasmas and other pathogens.

The invention discloses specific PCR primers used for detecting mycoplasma capricolum subsp. capripneumoniae. According to the specific PCR primers used for detecting the mycoplasma capricolum subsp.capripneumoniae, firstly, by conducting gene sequencing and comparing on the mycoplasma capricolum subsp. capripneumoniae, a target sequence used for detecting the mycoplasma capricolum subsp. capripneumoniae is screened out, a gene sequence of the target sequence is shown in SEQ ID NO.1 as shown in the description, then the specific PCR primers used for detecting the mycoplasma capricolum subsp.capripneumoniae are designed, the specific PCR primers used for detecting the mycoplasma capricolum subsp. capripneumoniae can distinguish the Mccp (mycoplasma capricolum subsp. capripneumoniae) fromother members of mycoplasma capricolum clusters and common ruminant mycoplasmas, show good specificity, and can be used as a novel diagnosis target, and on the basis of the specific PCR primers used for detecting the mycoplasma capricolum subsp. capripneumoniae, an Mccp diagnosis technology with higher clinical practicability is built.

The invention provides a traditional Chinese medicine tablet for treating Mycoplasma gallisepticum infection and a preparation method of the traditional Chinese medicine tablet, and belongs to the technical field of veterinary medicine. According to the traditional Chinese medicine tablet for treating the Mycoplasma gallisepticum infection and the preparation method of the traditional Chinese medicine tablet, drugs with heat clearing, toxicity removing, and infectious disease treating effects such as radix isatidis and flos lonicerae are used as main drugs, radix scutellariae has broad-spectrum antibacterial and antiviral effects, prevents secondary or concurrent bacterial infections, and is a ministerial drug in a prescription, radix glycyrrhizae has spleen tonifying, qi benefiting, lungmoistening, cough relieving, heat clearing, detoxifying, and medical nature regulating functions, aster has lung moistening, qi descending, inflammation diminishing, and cough relieving functions, andthe radix glycyrrhizae and the aster are both supplementary drugs; and the various drugs are jointly used for realizing heat clearing, detoxifying, cough relieving, and asthma relieving effects together. According to the traditional Chinese medicine tablet, the effects on preventing and treating the Mycoplasma gallisepticum infection are good, imbalance of various aspects in the course of a disease can be improved and regulated, coordination of an internal environment and recovery of the disease are promoted, the multi-directional, multi-level, multi-target pharmacological characteristics ofa compound traditional Chinese medicine are given full play to, and the characteristics that residues are low, toxicity is low, the curing rate is high, and drug resistance is not prone to being generated are realized.

The invention relates to feed for pheasants which are 40-60 days old. The feed is prepared from corn, fish powder, wheat bran, bone powder, feather meal, artemisia seleirgensis, thesium chinense, feather cockscomb seeds, purslane, pine needle powder, laminaria japonica powder, lactoferrin, stachyose, sodium caseinate, calcium hydrophosphate, zeolite powder, clove oil, origanum oil and blood meal. The feed is balance in formula nutrient and can meet the nutrient requirements for normal growth of the pheasants which are 40-60 days old. The feed can also prevent the young pheasants from being infected by coccidiosis, mycoplasmosis and colibacillosis, and increases the growth speed of the pheasants which are 40 days old.

The invention belongs to the technical field of veterinary drugs, and particularly relates to a traditional Chinese medicine composition for preventing and treating mycoplasmal pneumonia of swine anda preparation method thereof. The traditional Chinese medicine composition is prepared from, by weight, 20-45 parts of herba ephedrae, 15-35 parts of apricot kernels, 10-40 parts of codonopsis pilosula, 20-45 parts of radix liquiritiae, 35-55 parts of gypsum, 10-25 parts of venenum bufonis, 15-40 parts of parched radix paeoniae alba, 25-40 parts of fructus schisandrae chinensis, 5-30 parts of radix scutellariae, 15-35 parts of radix bupleuri, 15-35 parts of earthworms, 10-35 parts of semen lepidii, 25-45 parts of polygala roots, 5-35 parts of willowleaf rhizomes, 10-35 parts of perilla seeds,15-35 parts of flos lonicerae, 20-40 parts of radix asteris and 15-40 parts of Chinese yams. The traditional Chinese medicine composition is a pure traditional Chinese medicine preparation, is reasonable in medicine compatibility, safe and effective, addresses both symptoms and root causes, is free of relapse, has good curative effects on prevention and treatment for mycoplasma pneumonia of the swine, has good effects on chronic respiratory infectious diseases, coughs and asthma, avoids medicine residues due to antibiotic use for prevention and treatment, ensures food safety, and is suitable for popularization in livestock production.

The invention relates to a combined inactivate vaccine for sheep pox and contagious caprine pleuropneumonia (CCPP), and a production method of the combined inactivate vaccine. The method comprises thesteps of inoculating suspension cultured BHK-21 cells and a proper culture medium with a capripox virus AV41 strain and a mycoplasma capricolum pneumonia subspecies C87001 strain respectively for culturing, harvesting the culture, concentrating and purifying; inactivating with BEI and thimerosal respectively, adding a suitable adjuvant according to a certain proportion, mixing and emulsifying. The combined inactivate vaccine is used for preventing the sheep pox and the CCPP. The combined inactivate vaccine, provided by the invention, for the sheep pox and the CCPP is obtained internationallyfor the first time, and can achieve the aim of preventing the two diseases by means of one injection; furthermore, a live vaccine for the sheep pox is produced by means of the suspension cultured BHK-21 cells; compared with the original inactivate vaccine prepared from ovine testicular cells, the combined inactivate vaccine provided by the invention has obvious advantages, and can solve the problems that in the production process of the live vaccine for the sheep pox, the sheet testis is difficult to obtain, lamb death and ewe abortion are easily caused, subcutaneous immunization needs to be performed when the vaccine is used, an operation is difficult during clinical application, and the like.

The invention discloses a test strip for testing mycoplasma capricolum subsp.capripneumonia and a preparation method of the test strip. The test strip is characterized in that an antigen enveloped by a colloidal gold pad is a mycoplasma capricolum subsp.capripneumonia whole cell antigen obtained after ultrasonic wave disruption, a test line envelops a capricolum subsp.capripneumonia polysaccharide antigen, and a quality control line is mycoplasma capricolum subsp.capripneumonia whole-cell poly-IgG (immunoglobulin G). The test strip can test antibodies generated by the mycoplasma capricolum subsp.capripneumonia really, has specificity of being not in cross reaction with any other similar mycoplasmas and has the advantages of low cost, simplicity in operation, long-term storage and no toxicity and harm to human bodies.

The invention belongs to the technical field of veterinary drugs, in particular to a traditional Chinese medicine composition for preventing and treating mycoplasmal pneumoniae in sheep and a preparation method of the composition. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 15-40 parts of asterageratoides, 20-45 parts of herba hyperici monogyni, 15-40 parts of herba macleayae cordatae cum radice, 10-35 parts of acanthopanax evodiaefolius franch, 25-45 parts of folium livistonae chinensis and 10-35 parts of ramulus et folium picrasmae. The composition is a pure traditional Chinese medicine preparation, has the effects of clearing away heat and toxic materials, relieving cough and asthma, resisting bacteria and diminishing inflammation, the traditional Chinese medicine composition is reasonable in compatibility, safe and effective, treats both symptoms and root causes, has no relapse, avoids drug residues prevented and treated by antibiotics, ensures food safety, is suitable for popularization in animal husbandry production, and has wide market prospect and larger economic benefits.

The invention relates to application of a medicinal composition in preparation of a medicine for treating pneumonia. The composition is prepared from the following traditional Chinese medicine materials in parts by weight: 26-40 parts of herba houttuyniae, 26-40 parts of rhizoma belamcandae, 18-30 parts of cacumen biotae, 15-25 parts of radix scutellariae, 18-30 parts of common ducksmeat herb, 18-30 parts of dried tangerine peel, 8-16 parts of herba ephedrae and 8-16 parts of cortex moutan. The medicinal composition can be used in match with erythrocin to treat mycoplasma pneumonia in childrenand is capable of effectively reducing the time of abatement of fever, the cough eliminating time, and the lung noise eliminating time of the children, and thus the pain of children patients can be relieved.

The invention discloses a genetic engineering subunit vaccine of goat contagious pleuropneumonia as well as a preparation method and application thereof. The vaccine comprises a first recombinant protein, a second recombinant protein and a pharmaceutically acceptable carrier, wherein the first recombinant protein and the second recombinant protein respectively have sequences as shown in SEQ ID NO: 2 and SEQ ID NO: 6. The vaccine provided by the invention is free of any toxicity, high in safety and good in immunogenicity, and can generate relatively strong humoral immunity in a sheep body, antigens expressed by the first recombinant protein and the second recombinant protein have a very good cross protection effect, which can simultaneously resist attacks of mycoplasma capricolum subsp.capripneumoniae, mycoplasma capricomlusubsp.capricolum and mycoplasma ovipneumoniae, the immunized sheep can resist virulent virus attack, and the vaccine can be prepared through large-scale serum-free suspension culture by using a bioreactor, and has advantages of easy quality control, batch-to-batch stability, low production cost and the like.

The invention discloses a culture medium and fermentation culture method used for high-density culture of mycoplasma capricolumsubsp. capripneumonia. The mycoplasma capricolumsubsp. capripneumonia isisolated from respiratory secretions of a goat clinically diagnosed with mycoplasmal pneumonia; the mycoplasma capricolumsubsp. capripneumonia has been collected in China General Microbiological Culture Collection Center on 4 December, 2019, and the collection number is CGMCC No.18878; according to the fermentation culture method, liquid fermentation culture is carried out on the resuscitated-passaging mycoplasma capricolumsubsp. capripneumonia ZFY17 CGMCC No.18878 in a specific fermentation culture medium under specific conditions; and the specific fermentation culture medium replaces part ofhorse serum with a combination of glucose, hydrolyzed yeast protein, a hydrolyzed yeast culture, lactoalbumin hydrolysate and casein, thus greatly reducing strain fermentation cost.

The invention discloses an iELISA method for screening serologically negative sheep of Mycoplasma ovis pneumoniae and Mycoplasma capricoides subspecies capneumoniae. , the bacterial solution was centrifuged, sterilized, and ultrasonically resuspended in PBS to separate the antigen; (2) Coat the microtiter plate with the antigen, incubator at 37°C for 1 hour, then overnight at 4°C; add blocking solution, block at 37°C for 2 hours; add Serum samples diluted 200 times were incubated at 37°C for 1 hour; rabbit anti-goat secondary antibody diluted 5000 times with 5% skimmed milk powder was added, and incubated at 37°C for 1 hour; (3) After washing the plate with PBST, add TMB chromogenic solution and incubate at 37°C 10min; add stop solution to terminate the reaction; read OD with microplate reader 450nm value. The sensitivity of the iELISA method of the invention can reach 98%, the operation is simple, and the sample processing capacity is large, which can not only be used for the screening of Mccp and Mo two pathogenic serologically negative sheep, but also be used for monitoring the level of Mccp antibody in the experiment.