1. 称取42.5 g本品,加1 L去离子水溶解。
2. 边搅动边加热,煮沸1min,使其完全溶解。
3. 121ºC灭菌15min。
产品用法: 称取本品 42.75g,加热搅拌溶解于 1000ml 蒸馏水中,分装试管,121℃高压灭菌 15 分钟备用。
强化梭菌鉴别琼脂(DRCA)实验方法:
1. Prepare serial 10-fold dilutions of the sample in 1/4 strength Ringer’s solution or 0.1% peptone water.
2. Depending on the amount of the initial sample, transfer 1 mL or 0.1 mL of the appropriate dilution, prepared in step 1, to the bottom of a molten (45-50°C) DRCA tube.Prepare a duplicate tube using the same procedure.
3. Tighten the caps on the tubes.
4. Heat one of the duplicate DRCA tubes prepared in step 2 to 80 ± 1°C for 10 minutes to kill vegetative cells.
5. Incubate both tubes, heat-shocked and non-heat-shocked, at 35 ± 1°C for 5 days; examine for sulfite reduction. Non-heat-shocked cultures showing blackening must be heat shocked and subcultured to DRCA for confirmation.
其它方法
Inoculate samples onto the surface of agar plates using the streak plate or spread plate technique. Samples may be inoculated into DRCA using the pour plate technique. Agar in agar deeps may be inoculated using the stab technique. DRCA may be used to overlay the membrane filter in the membrane filter technique. Incubate plates and tubes at 35 ± 1°C for 24-48 hours under anaerobic conditions. Agar deeps may be incubated under aerobic conditions when following the Prickett tube method.
结果分析
The presence of clostridia is presumptively indicated by blackening in the Agar. Heat-shocked tubes showing blackening should be considered confirmatory for the presence of sulfite-reducing clostridia.