环介导等温扩增技术检测酸奶中嗜酸乳杆菌

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 r RNA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/m L,人工污染酸奶样品的检出限为120 CFU/m L,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。

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

To establish of loop mediated isothermal amplification (LAMP) method detect thermophilic Lactobacillus in yogurt. Method Based on the GeneBank in 16S-23S rRNA as target sequence design primer and optimize the LAMP reaction system and produce. At the same time, primer specificity was verified and the sensitivity and detection limit of Lactobacillus acidophilus were confirmed. Result The reaction results were determined by ladder strip through gel electrophoresis and white precipitate of magnesium pyrophosphate after centrifugation. Detection time is 5.5h . The sensitivity and detection limit by artificial contamination were 6.2×101 CFU/mL and 1.2 ×102 CFU/mL, respectively. For the specific detection of Lactobacillus acidophilus, 12 strains of Lactobacillus acidophilus bacteria specific results were 100%, which showed the primer with high specificity. Conclusion LAMP could be used to detect Lactobacillus acidophilus in yogurt with high sensitivity and specificity, sipmle operation and less time.