环介导等温扩增技术检测酸奶中嗜酸乳杆菌

Rapid detection of Lactobacillus acidophilus in yogurt by loop-mediated isothermal amplification

中文关键词:  环介导等温扩增法  嗜酸乳杆菌  酸乳

英文关键词:loop-mediated isothermal amplification  Lactobacillus acidophilus  yogurt

中文摘要:

      目的 建立环介导等温扩增法(loop-mediated isothermal amplification, LAMP)检测酸奶中嗜酸乳杆菌。方法 以嗜酸乳杆菌16S rRNA保守区的基因序列为靶序列设计引物, 优化建立LAMP反应体系和程序, 并研究引物的特异性, 同时确定嗜酸乳杆菌的灵敏度和检出限。结果 本方法可通过凝胶电泳和离心后焦磷酸镁白色沉淀检测反应结果, 检测时间为5.5 h, 灵敏度为62 CFU/mL, 人工污染酸奶样品的检出限为120 CFU/mL, 12株非嗜酸乳杆菌细菌非特异检测结果均为100%。结论 该方法灵敏性好、特异性强、操作简单、耗时短, 可适合于酸奶中的嗜酸乳杆菌的快速测定。

英文摘要:

      Objective To establish a method for detection of thermophilic Lactobacillus in yogurt by loop mediated isothermal amplification (LAMP). Methods Using the 16S rRNA of Lactobacillus acidophilus as target sequences, the primers were designed and the LAMP reaction system and produce were built and optimized. At the same time, the specificity of primers was verified and the sensitivity and detection limit of Lactobacillus acidophilus were confirmed. Results The reaction results were determined by ladder strip through gel electrophoresis and white precipitate of magnesium pyrophosphate after centrifugation. Detection time was 5.5 h. The sensitivity and detection limit by artificial contamination were 62 CFU/mL and 120 CFU/mL, respectively. The specific results of 12 strains of Lactobacillus acidophilus bacteria were 100%, which showed the primers had high specificity. Conclusion LAMP can be used to detect Lactobacillus acidophilus in yogurt with high sensitivity, high specificity, simple operation and short time consuming.