Optimization of pre-enrichment protocol of PCR-based method for detection of Escherichia coli
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摘要: 目的:比较不同培养模式与不同添加剂对大肠杆菌增殖效果的影响。方法:本实验以大肠杆菌为目标菌,研究了2种培养模式(振荡与静止培养)、2种添加剂(甘露醇与乳糖)对大肠杆菌增殖效果的影响,并采用PCR法检验其增殖效果。结果得出:振荡培养的增殖效果优于静止培养。在LB培养基中添加甘露醇和乳糖可起增殖作用,其中0.3g/100mL甘露醇与1.0%乳糖复配4h后,大肠杆菌的增殖效果最佳。检测中使用0.3g/100mL的甘露醇和1.0%的乳糖,并辅以37℃、150r/min的振荡培养即可获得良好效果。结论:本方法在6h内即可完成大肠杆菌的增殖及PCR检测,可用于食品中大肠杆菌的快速检测。Abstract: Objective: To compare the enrichment effect of different culture methods and different medium additives on Escherichia coli. Method: This study was to investigate the enrichment effect of 2 culture methods ( static cultivation and shaking cultivation) and 2 medium additives ( lactose and mannitol) on Escherichia coli, and to evaluate the enrichment effect by PCR.Results: The results indicated that shaking cultivation was better than static cultivation.The effect of LB medium adding mannitol and lactose was beneficial to the growth of Escherichia coli.When the dosage of lactose and mannitol were 1.0% and 0.3g /100 mL, the effect of enrichment was best. The enrichment time was 4h.The detection could get good results by using 1.0% lactose and 0.3g /100 mL mannitol with shaking cultivation at 37℃.Conclusion: The enrichment and PCR detection of Escherichia coli can be accomplished in 6h, which can be used for rapid detection of Escherichia coli in foods.
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Keywords:
- PCR detection /
- Escherichia coli /
- pre-enrichment
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