Strain Anaerostipes sp B2131 and application thereof in inflammatory intestinal diseases
A technology of inflammatory bowel disease, cgmccno.1.5295, applied in the field of microorganisms, can solve the problem of lack of Lachnospiraceae strains and other problems, achieve significant application value, have no toxic side effects, and improve the effect of intestinal flora
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[0040] Reagents, materials and equipment
[0041]
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Embodiment 1
[0043] Embodiment 1, the isolation, cultivation and identification of Anaerostipes sp.B2131 bacteria
[0044] (1) Conditioned medium preparation
[0045] Pre-deoxygenated liquid BHI medium (containing 5% bovine serum and 0.1% cysteine): Weigh 37 g of brain heart infusion (BD company) into a 1 L Erlenmeyer flask, add 1 g of cysteine and double distilled Dilute the water to 1000ml, stir to dissolve, and then sterilize in an autoclave at 121°C for 15min; Shake evenly to obtain pre-deoxygenated liquid BHI medium, and then divide into 50ml centrifuge tubes.
[0046] Pre-deoxygenated solid BHI medium (containing 5% bovine serum, 0.1% cysteine and 1 mg / L aztreonam): take 37 g of brain heart infusion (BD company) in a conical flask with a capacity of 1 L, add 1g cysteine, 15g agar powder and double-distilled water to a volume of 1000ml, stir to dissolve, and then sterilize in an autoclave at 121°C for 15min; after the sterilization is completed, put the medium into an anaerobic ...
Embodiment 2
[0063] Example 2, Abundance Detection of Anaerostipes sp. Bacterial Content in IBD Patients and Healthy Controls
[0064] (1) Fecal flora DNA extraction
[0065] Stool samples from IBD patients were provided by inpatients in the Inflammatory Bowel Disease Center of Run Run Shaw Hospital Affiliated to Zhejiang University. After collection, the samples were frozen and stored in a -20°C refrigerator in time, and transported to the laboratory with dry ice in a -80°C refrigerator within 2 days. The fecal samples of healthy people were provided by healthy people for physical examination, and they were taken back to the laboratory within 5 hours and stored in a -80°C refrigerator. When extracting fecal DNA, take out the sample from -80°C, and take 100-150 mg of feces with a medicine spoon into a 1.5ml centrifuge tube without thawing; then, use the fecal flora DNA extraction kit (Qiagen) to extract the total DNA of fecal flora , and then use the Nanodrop 2000 UV micro-spectrophotomet...
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