Genitourinary tract mycoplasma culture medium and detect method thereof

A technique for urogenital tract and detection method, which is applied in the field of urogenital tract microorganism culture and detection, can solve the problems of no mycoplasma culture medium and preparation method, the culture medium affects the detection effect, the culture medium is expensive, and the like, and achieves typical colony morphology. , the effect of prolonging the logarithmic phase of growth and reducing the false negative rate

Inactive Publication Date: 2009-10-14
上海市皮肤病性病医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the growth of urease-producing bacteria leads to acid-base changes, which can also cause misjudgment of Uu liquid culture results [Hebei Medical Sciences 2004, 10(7): 614-616]
[0005] Solid culture is used to identify various mycoplasmas by observing the presence or absence of oil-fried egg-like colony growth and morphological characteristics under a microscope, but the quality of domestic solid culture medium is far behind that of imported A7 solid culture medium
Imported solid medium is expensive, the order cycle is long, and the medium becomes dry and thin, which may affect the detection effect
In addition, directly inoculating the sample into solid medium has a lower detection positive rate
Clinical practice has found that a large number of commercially used liquid or solid culture media for Mycoplasma have great differences in sensitivity and specificity. Even if the positive rate of liquid culture of the same brand is significantly higher than that of solid culture, there is currently no high-quality culture medium. Mycoplasma culture medium and preparation method used in combination

Method used

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  • Genitourinary tract mycoplasma culture medium and detect method thereof
  • Genitourinary tract mycoplasma culture medium and detect method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Culture medium configuration: Take a 2000ml clean small-bore, high-pressure resistant glass container with a cover (pluggable), weigh 5g~10g of trypsin soybean broth dry powder and dissolve it in 600ml of purified water, add NaCl and diphosphate Potassium hydrogen, adjust to pH 6.1~6.3 with 1N HCL, paste disinfection and sterilization instruction strips, place in a high-pressure container, sterilize at 121°C for 15 minutes after removing cold air, and remove residual steam after the pressure returns to zero. Cover and cool naturally for later use.

[0068] Take a clean container and add an appropriate amount of sterilized purified water, weigh yeast extract, urea, arginine, L-cysteine, and phenol red into the container one by one according to the formula, pressurize and pass through a 0.22μm filter , add the solution to the cooled above-mentioned culture medium, take a sterile container and inject an appropriate amount of sterilized water, weigh vancomycin, amphotericin...

Embodiment 2

[0071] The configuration of the solid medium: take a 2000ml clean small-caliber, high-pressure-resistant thick-walled glass container with a cover (can be plugged), add about 500ml of purified water, and weigh trypsin soybean broth dry powder, agar powder, Dissolve manganese sulfate into the container one by one, adjust to pH 6.1~6.3 with 1N HCL, paste the disinfection and sterilization instruction strip, put it in a high-pressure container, and sterilize at 121°C for 15 minutes after excluding cold air, and wait until the pressure returns to zero Finally, remove the residual steam and open the cover to cool naturally to 56°C, and keep it warm for later use.

[0072] Take a clean container and add an appropriate amount of sterilized purified water, weigh the yeast extract, urea, arginine, L-cysteine, and phenol red into the container one by one to dissolve, pass through a 0.22 μm filter, and then pour into the insulated culture In the base, take a sterile container and inject ...

Embodiment 3

[0075] Comparison of the detection ability of different culture media on clinical samples: take swabs of clinical samples, put them into sterile small test tubes containing 50 μl to 200 μl of normal saline, discard the swabs after fully stirring, and add the liquid samples equally to the present invention , imported A and domestic B liquid medium, at 37 ℃, 5% CO 2 Cultivate under the conditions for 24 hours, and replant in the solid medium of the present invention and import A respectively, because domestic B commodity does not have corresponding solid medium, only compare the performance of liquid medium. Liquid and solid media containing samples to be tested, continue to store at 37°C, 5% CO 2 conditions, up to 72 hours. Observe the following indicators of the medium every 12 hours: whether the liquid medium changes color, time, turbidity, solid culture colony growth rate, in order to understand the positive rate of culture, pH buffer capacity, record the changes in the thr...

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Abstract

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.

Description

technical field [0001] The invention relates to a method for cultivating and detecting microorganisms in the urogenital tract, in particular to a culture medium and a detection method for ureaplasma urealyticum (Uu) and mycoplasma hominis (Mh) in the urogenital tract. Background technique [0002] Mycoplasma is the smallest microorganism known to be able to grow independently and reproduce by itself. It has no cell wall, is widely distributed, and can grow on artificial medium, but requires high nutritional conditions. They are closely related to urogenital inflammation and can also cause immune diseases [Mycoplasma, 2nd Edition, 2008, 41-44]. Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) have high incidence and strong pathogenicity. They are related to female reproductive tract infections, such as cervicitis, endometritis, pelvic inflammatory disease, and some complications such as spontaneous abortion, premature rupture of membranes, and premature delivery. Uu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04C12R1/01
Inventor 顾伟鸣吴磊杨阳
Owner 上海市皮肤病性病医院
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