58 results about "Mycoplasma culture" patented technology

The invention relates to an efficient mycoplasma hyopneumoniae culture medium and a preparation method thereof, and belongs to the technical field of veterinary biology. The efficient mycoplasma hyopneumoniae culture medium comprises an A liquid and a B liquid mainly consisting of MEM, yeast leaching powder, tryptone, glucose, inorganic salt and the like. The prepared culture medium of the invention has the main advantages of low serum content which is only 10%-15%, while the serum content in common culture medium is 20% even more. The culture medium prepared by the low serum relieves the pig allergic to the stress reaction, meanwhile gives consideration to the biosafety of animals. Besides the valence of the semi-finished bacterial solution prepared by the method is up to 109CCU/ml, which is much higher than the culture medium prepared by the common technology.

The invention relates to a culture medium adopting continuous enrichment culture for mycoplasma gaujseptium and a preparation method, belonging to the technical field of veterinary biology. The culture medium comprises PPLO (pleuropneumonia-like organism), yeast extract powder, glucose and the like, wherein before use, healthy pig serum is added. The culture medium can ensure the high titer and stability of the growth of a semi-finished product, and the semi-finished product bacterium solution prepared by the method disclosed by the invention is up to 10<12> CCU/ml; simultaneously, the culture medium is used for culturing mycoplasma gaujseptium by adopting a continuous enrichment culture process, so that the stable growth phase of the propagated thalli of mycoplasma gaujseptium is prolonged, the active ingredients in the culture medium are furthest utilized and consumed, and then the active antigens of mycoplasma gaujseptium are greatly increased and incremented; and the final bacterium solution can be used for preparing an inactivated vaccine directly or after being diluted, and does not need to be concentrated, so that the production process is simplified, and the production cost is reduced.

The invention relates to a mycoplasma hyopneumoniae culture medium and a preparation method thereof. The mycoplasma hyopneumoniae culture medium consists of a basic culture medium and an auxiliary culture medium, wherein the basic culture medium has major components of Hank's liquid, lactoprotein hydrolysate, a yeast extract and an ox heart extract, sterilization can be performed through autoclaving, and a pollution risk caused by filtration sterilization is greatly reduced; and the auxiliary culture medium comprises pig serum, argenine, cysteine, phenol red solution, penicillium and the like, the pig serum is sterilized through cobalt radiation, and the rest components are mixed with inactivated pig serum after filtration sterilization. The titer of a lapinized attenuated strain of the mycoplasma hyopneumoniae cultured with the culture medium provided by the invention is 109-1010 CCU, the minimum culture time may be 40 h, the generation of old bacteria and aged bacteria is greatly reduced, the usage amount of the pig serum can be as low as 8%, the allergic stress reaction caused by the pig serum is reduced, and the production cost of an enterprise is lowered.

The invention discloses a mycoplasma hyopneumoniae culture medium and a preparation method thereof, which belong to the technical field of veterinary biological products. Each 1000 ml of phosphate buffer contains 20-50 g of PPLO (pleuropneumonia-like organism), 2-10 g of yeast powder, 3-15 g of glucose, 0.5-4 ml of 1% phenol red, 2-10 g of amino acid composition, 15-30 g of compound traditional Chinese medicine polysaccharide and 50-100 ml of pig serum. When culturing the mycoplasma hyopneumoniae, the mycoplasma hyopneumoniae culture medium disclosed by the invention can improve the tolerance of the mycoplasma hyopneumoniae and reduce the apoptosis rate of the mycoplasma hyopneumoniae; the mycoplasma hyopneumoniae culture medium disclosed by the invention can increase the amount of culture bacteria of the mycoplasma hyopneumoniae, the concentration of the culture bacteria liquid reaches 10<10-11>/ml, which is increased by more than 5-10 times compared with that of a common culture medium; the mycoplasma hyopneumoniae culture medium disclosed by the invention can inhibit the growth of other bacteria when culturing the mycoplasma hyopneumoniae so as to reduce the culture pollution risk of culturing the mycoplasma hyopneumoniae.

The invention relates to a combined culture reagent box of Candida Albicans, hemophilic bacteria and mycoplasma and application thereof. The reagent box comprises a culture medium and a CO2 generator. The culture medium comprises culture medium of Candida Albicans, culture medium of hemophilic bacteria and culture medium of mycoplasma. The pH value of the culture medium is 6.3 plus or minus 0.5. The invention is used to detect Candida Albicans, hemophilic bacteria and mycoplasma, also evaluate the microecology of the urogenital tract of male and female, diagnose the infection of Candida Albicans, hemophilic bacteria and mycoplasma, and evaluate the curative effect of the bacteria. Compared with the prior art, the combined culture reagent box of Candida Albicans, hemophilic bacteria and mycoplasma of the invention has the advantages of the simple operation, the reliable method, the stable characteristics, and easily observing and judging the results, provides a diagnostic tool with more scientific, more convenient, more humanized and more accurate, is favorable for biological or medical practice and commercialized transportation and has the good social and economic efficiency.

The invention provides a mycoplasma anatis culture medium. The mycoplasma anatis culture medium is composed of a liquid culture medium and a solid culture medium. The liquid culture medium is prepared from deionized water, a 10% thallium acetate solution, PPLO broth, glucose, peptone, tryptone, 1% phenol red, a CMRL-1066 culture medium containing L-glutamine, inactivated horse serum, yeast leachate, penicillin, L-cysteine hydrochloride and nicotinamide adenine dinucleotide. Besides the situation that agar powder is added and phenol red is removed, other components of the solid culture medium are the same as those of the liquid culture medium. The culture medium is used for separating and purifying mycoplasma anatis, wherein a bacterial colony is separated from the solid culture medium, the separated bacterial colony is placed in the liquid culture medium to grow, then filtering, dilution and plate coating of liquid culture substances and purification of picked single colony multiplication are conducted, and the steps are repeated till purified mycoplasma anatis is obtained. The culture medium can promote growth of mycoplasma anatis, and the success rate of separation and purification of mycoplasma anatis is high.

The difficult problem of the contamination of cell cultured mycoplasma and the infection diagnosis of various mycoplasma urgently need a reliable and effective kit. Mycoplasma culture by a fluorescent staining method is reliable but takes time and trouble, and culture pollution is intensified. A direct PRC method for amplifying a mycoplasma gene, which is used for the routine detection of the mycoplasma is easy for cross contamination, and high false masculinity is only used as an auxiliary reference. The invention relates to a mycoplasma conservative gene which across links a plant arabidopsis LEY sequence into a reporting gene DNA by a hybridization probe and amplifies the indirect detection of a reporting gene. The mycoplasma conservative gene is characterized in that mycoplasma 16sRNA is hybridized with the head part and the tail part of the LEY reporting gene DNA whose head part and tail part are provided with mycoplasma conservative sequence probes; the reporting gene DNA is catalyzed into a ring by heat-proof ligase, and the annular reporting gene is reversely amplified by PCR so as to indirectly reflect the detection of the mycoplasma; by adjusting ligase reaction, the reporting gene is difficultly across linked by the cross contamination, thereby reducing the false masculinity; and if a system is contaminated, the reporting gene can be changed.

The invention relates to genitourinary tract microorganism cultivation and detect method thereof, in particular to a genitourinary tract ureaplasma urealyticum (Uu) and mycoplasma hominis (Mh) culture medium and a detect method thereof. The culture medium comprises a fluid medium and a solid medium which are combined when the medium is used for culturing, separating and differentiating mycoplasma. The formulation comprises essential elements such as tryptic soy broth, yeast extract, urea, arginine, L-cysteine, phenol red, HEPES liquid, calf (fetal calf) serum, manganese sulfate, vancomycin, amphotericin B, trimethoprim, polymyxin, penicillin, and the like, and thereby the nutrition for mycoplasma growth and the antibiotic combination for inhibiting undesired microbe growth are optimized, no millipore filter is needed for sample inoculation and subcultivation, and the mycoplasma grows quickly. The fluid medium can maintain and prolong the logarithmic phase and indicate the growth of the mycoplasma, and the solid medium can used for observing Uu and Mh bacterial colonies, which has large background reflectance, thereby being easy for differentiation and significantly increasing the sensibility and the specificity of genitourinary tract mycoplasma detect.

The invention discloses a mycoplasma ovipneumoniae low-serum medium, which consists of the following components: a PPLO broth, sodium pyruvate, phenol red, de-ionized water, lactose, tryptone, a fresh yeast extract, insulin, L-glutamine, L-cysteine, lactoalbumin hydrolysate, transferrin, penicillin and little horse serum; and the invention provides a preparation method of the mycoplasma ovipneumoniae low-serum medium. The mycoplasma ovipneumoniae medium provided by the invention has the following beneficial effects: a serum content is merely 5%, which is 1/4 of the serum content of a medium in the prior art, and the living bacterium titer of cultivated mycoplasma ovipneumoniae reaches 7*10<9> CCU/mL or above, which is significantly higher than that of the medium in the prior art. In comparison with the prior art, the mycoplasma ovipneumoniae low-serum medium provided by the invention has the characteristics of being low in serum content, rapid in growth and high in living bacterium titer.

The invention discloses a bovine mycoplasma Taqman fluorescence PCR kit and a using method thereof. The kit comprises a TaqMan probe, a PCR detection primer or a derived sequence of the primer, PCR amplification reagent, positive control and negative control. Each reagent is added according to the using method to form a reaction system, and is used for detecting a sample according to the fluorescence PCR instrument reaction condition set according to instruction of the using method, and can be used for quickly detecting the bovine mycoplasma nucleic acid in a bovine mycoplasma culture or tissue sample. The kit can be used for quickly detecting a bovine mycoplasma pneumonia suspected case on a cattle farm, and has the characteristics of simplicity, accuracy, quickness and the like.

The invention belongs to the technical field of animal quarantine, and particularly relates to a chicken poison mycoplasma fluorescent quantitative detection kit and a detection method thereof. The detection kit comprises a fluorescent quantitative PCR reaction liquid, a positive control and a negative control, wherein the fluorescent quantitative PCR reaction liquid comprises an upstream primer, a downstream primer and SYBR Premi*E*Taq (Tli RNaseH Plus). By adopting the detection kit to conduct chicken poison mycoplasma fluorescent quantitative PCR detection on samples, the preprocessing process of the samples is very simple, there is no need to conduct mycoplasma cultivation on the samples or conduct mycoplasma gene extraction on the samples, and cooling after direct boiling of the samples is only required, so that both time and cost are saved. The detection kit has the advantages of being high in detecting sensitivity, good in specificity and repeated stability, and is very applicable to the detection of the chicken poison mycoplasma in the chicken throat swabs.

The invention discloses a lower-serum and efficient culture medium of mycoplasma ovipneumoniae. The lower-serum and efficient culture medium is prepared from the following components: MEM, sodium pyruvate, glucose, Hank's liquid, fresh yeast leaching liquid, L-glutamine, L-cysteine, lactalbumin hydrolysate, calf thymus DNA (Deoxyribonucleic Acid), insulin, transferrin, penicillin, horse serum, phenol red and de-ionized water; the invention provides a preparation method of the lower-serum and efficient culture medium. The lower-serum and efficient culture medium of the mycoplasma ovipneumoniae, disclosed by the invention, has the beneficial effects that the content of the serum is only 5 percent and is 1/4 of the content of the serum in a culture medium in the prior art; the titer of mycoplasma ovipneumoniae semi-finished-product bacterium liquid prepared by the method reaches 10<10>CCU/ml and is obviously higher than that of the culture medium in the prior art. According to the lower-serum and efficient culture medium of the mycoplasma ovipneumoniae, disclosed by the invention, the sensitive stress response to a goat body, caused by heterologous serum, is alleviated, and the biosafety is improved; the titer of bacterium liquid of living bacteria is also improved and the production cost is reduced.

The invention provides a low-serum efficient culture medium for culturing mycoplasma hyopneumoniae and a preparation method thereof. The low-serum efficient culture medium comprises a base medium and an addition bag. The base medium comprises M199, brain extract powder, lactoalbumin hydrolysate, BSA, glucose, fresh yeast extract, Hepes, iron citrate, calcium chloride, 2H2O, phenol red and dd H2O. The addition bag comprises cholesterol, linoleic acid, linolenic acid, palmitic acid, ethanol and porcine serum. Through use of some hydrolyzates and lipid, a serum addition ratio is less than 5% so that a cost is reduced, the quality of mycoplasma is improved, and mycoplasma hyopneumoniae antigen titer is improved.

The present invention relates to a preparation method of an egg yolk antibody injection for treating porcine respiratory syndrome, which comprises the following steps: (1), isolating the current prevalent PRRSV (porcine reproductive and respiratory syndrome virus), PCV-2 (porcine circovirus virus) and mycoplasma hyopneumoniae, and culturing the PRRSV with the MARC-145 cells, the PCV-2 with the PK15 cells and the mycoplasma hyopneumoniae with the mycoplasma culture medium respectively to prepare the triple inactivated vaccine after measuring the titer of virus and mycoplasma; (2), immunizing the nonimmune laying hen with the prepared triple inactivated vaccine for PRRSV/PCV2 and mycoplasma hyopneumoniae; (3), collecting the egg yolk in the aseptic conditions; (4), measuring the valence of neutralizing antibody in the PRRSV and PCV-2 through microneutralization experiment and measuring the valence of antibody of in the mycoplasma hyopneumoniae through ELISA (enzyme-linked immune sorbent assay); and (5), performing sterility test for the egg yolk after valence measurement and performing sterile packaging after testing qualified. The preparation method has the benefits of effectively treating pigs which are independently infected or mixedly infected with PRRSV, PCV2 and mycoplasma hyopneumoniae diseases.

The invention provides a method for separating mycoplasma ovipneumoniae, which comprises the following steps: (1) collecting a sample and treating the sample to obtain a sample inoculation solution; (2) inoculating chick embryos; (3) separating the mycoplasma ovipneumoniae: collecting dead chick embryos after inoculation and/or non-dead chick embryos after 9 days, taking chick embryo allantoic fluid to carry out viable bacteria detection on the mycoplasma ovipneumoniae, and if the chick embryo allantoic fluid is collected to be positive, storing the material, if the chick embryo allantoic fluid of the first generation is negative, the collected chick embryo allantoic fluid is blindly transmitted to the chick embryo by the same method, and if the chick embryo allantoic fluid is still negative after three generations of blind transmission, determining the sample to be negative for mycoplasma ovipneumoniae, and discharging the material. The method disclosed by the invention is simple to operate, does not need to prepare a mycoplasma ovipneumoniae culture medium with complex components, and avoids the problem that in the prior art, high-pressure sterilization or a high-capacity filteris often needed in preparation of an isolation culture medium; and according to the method, the mycoplasma ovipneumoniae without animal serum can be obtained.

The invention belongs to the field of medical detection instruments and particularly relates to a mycoplasma culture plate automatic interpreter. The mycoplasma culture plate automatic interpreter comprises a culture assembly, an analysis assembly, a moving assembly, a main controller, a display screen, a printer and a shell carrying the above assemblies. For the mycoplasma culture plate automaticinterpreter provided by the invention, by arranging the culture assembly, the analysis assembly, the moving component, the main controller, the display screen and the printer, operation of artificialculture and manual interpretation in traditional operation is replaced, the working efficiency is greatly improved, and automatic operation of mycoplasma identification and drug sensitivity analysisis achieved.

The invention relates to the technical field of medical detection, in particular to a full-automatic mycoplasma culture sample-adding detector and a culture sample-adding detection method thereof, Thesample-adding detector comprises a consumable storage area, a working area, a cultivation area and a result interpretation area, and a work table is arranged at the bottom of the working area, a mechanical arm and a suction head are arranged above the work table, a specimen holder is arranged on one side of the work table, the specimen holder is provided with test tubes, the top of the test tubeis provided with a sample collection swab body card, the specimen collection swab body card is matched with a specimen holder concave card disposed at the upper part of the specimen holder, the test tube is also provided with a specimen collection swab head card, and the bottom of the sample collection swab head card is connected to a specimen collection swab rod. Compared with the prior art, thedetector can realize the automatic operation of the culture of the mycoplasma, the automatic operation of the instrument is realized, the reproducibility of the result is good, and the work efficiencyis improved.

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.

The invention provides a mycoplasma anseris in-vitro liquid culture medium. The mycoplasma anseris in-vitro liquid culture medium consists of a culture medium A and a culture medium B, wherein the culture medium A is a mycoplasma culture medium for breakdown of glucose; the culture medium B is a mycoplasma culture medium for breakdown of arginine; the culture medium S comprises the following ingredients: mycoplasma broth base, deionized water, glucose, sodium pyruvate, 1% phenol red, a 10% thallium acetate solution, inactivated horse serum, a yeast extract, penicillin, an L-cysteine hydrochloride aqueous solution and a nicotinamide adenine dinucleotide nad aqueous solution; the culture medium B comprises the following ingredients: mycoplasma broth base, deionized water, sodium pyruvate, 1% phenol red, a 10% thallium acetate solution, inactivated horse serum, a yeast extract, penicillin and an L-arginine monohydrochloride aqueous solution. By adoption of the mycoplasma anseris in-vitro liquid culture medium, growth can be realized only after 24 h, and the tilter after 48-h growth can be 10<3> CCU/mL. The mycoplasma anseris in-vitro liquid culture medium is suitable for isolated culture and diagnosis of mycoplasma anseris.