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Reagent kit for culturing and differentiating pathogenic mycoplasma and preparing method thereof
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A mycoplasma and kit technology, which is applied in the field of medical testing and can solve problems such as inconvenience in clinical use
Inactive Publication Date: 2008-01-23
SHANGHAI EASTDIAGNO MEDICAL TECH
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Problems solved by technology
At present, the identification kits on the market are for cultivating a certain type of mycoplasma alone, which brings inconvenience to clinical use.
Method used
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Embodiment 1
[0019] A diagnostic kit capable of cultivating and distinguishing U.u, M.h and M.p is provided. It is mainly divided into two processes: kit preparation and test operation:
[0020] 1. Preparation of kits
[0021] (1) Basic medium configuration
[0022] 1. Prepare sulfamethoxazole (SMZ) solution with a concentration of 50-100 mg / ml; prepare trimethoprim (TMP) solution with a concentration of 10-50 mg / ml, and mix the two to obtain compound sulfamethoxazole solution ( SXT), cryopreservation;
[0023] 2. Prepare a phenol red solution with a concentration of 1-5%, and store it at low temperature;
[0024] 3. Prepare the additive solution, in which the concentration of L-cysteine is 1-10 mg / ml, the concentration of urea is 10.0-100.0 mg / ml, the concentration of glucose is 200-1000 mg / ml, sterile filter, and store at low temperature;
[0025] 4. Prepare medium broth, in which the concentration of PPLO medium is 10-100g / L, the concentration of yeastpowder is 10-100g / L, and it ...
[0038] This kit includes basic culture medium, M.h identification reagent and M.p identification reagent group.
[0039] 1. According to the basic medium formula in Table 1, prepare the basic medium through the following steps,
[0040] 1) Weigh 550-1000 mg of sulfamethoxazole (SMZ), 110-500 mg of trimethoprim (TMP), add 10-20 ml of DMSO, fully dissolve, subpackage, and store at low temperature;
[0041] 2) Dissolve phenol red in 1N NaOH, add 3 to 5 times the volume of sterile deionized water, and store at low temperature;
[0042] 3) Weigh 10-50g of PPLO medium, 5-20g of yeastpowder, add 300-800ml of deionized water, heat and mix, adjust the pH to 6.2-7.2, and sterilize at 121°C for 15min.
[0043] 4) Weigh 0.2-1.0g L-cysteine, 3.0-10.0g urea, 20-100g glucose, dissolve in 50-200ml deionized water, adjust the pH to 6.2-7.2, add 0.5-5g ampicillin and SMZ 1....
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Abstract
The invention pertains to the field of medical inspection, and relates to a reagent box and preparing method for the reagent box for culturing and distinguishing pathogenic mycoplasma, in particular to a method for culturing and distinguishing Ureaplasma urealyticum, mycoplasma hominis, and mycoplasmal pneumonia. The invention prepares basic culture medium based on the common features of clinically common pathogenic mycoplasma, which is applicable for direct culturing of mycoplasma hominis; prepares distinguishing reagent for mycoplasma hominis and mycoplasmal pneumonia based on the different culturing features of mycoplasma hominis and mycoplasmal pneumonia. After adding the distinguishing reagent for mycoplasma hominis or mycoplasmal pneumonia in the basic culture reagent, the culture medium is only applicable for the growth of mycoplasma hominis or mycoplasmal pneumonia, and culture and distinguish various pathogenic mycoplasma for human being. The invention further prepares the reagent box, which obviously makes convenience for the clinic detection.
Description
technical field [0001] The invention belongs to the field of medical examination, and relates to mycoplasma cultivation, identification and clinical examination, more specifically, to a kit for cultivating and identifying pathogenic mycoplasma and a preparation method thereof, and more specifically, to a Ureaplasma urealyticum (U.u. ), Mycoplasma hominis (M.h) and Mycoplasma pneumoniae (M.p) culture identification method. Background technique [0002] Mycoplasma is a kind of prokaryotic microorganism lacking cell wall, the size is generally between 0.3-0.5um, it is highly pleomorphic, and has various forms such as spherical, rod-shaped, filamentous, and branched. It's different from a cell, and it's different from a virus. There are many kinds of mycoplasma, which are widely distributed and cause considerable harm. Among the mycoplasmas that are pathogenic to humans, Mycoplasma pneumoniae (M.p), Ureaplasma urealyticum (U.u) and Mycoplasma hominis (M.h) account for the vast...
Claims
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Application Information
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