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PCR结合变性高效液相色谱快速检测发酵乳酸杆菌
杨大伟1,2, 周裔彬1, 刘云国2, 雷质文2, 陈俊芳1, 王建广2, 李正义2
1.安徽农业大学;2.山东出入境检验检疫局检验检疫技术中心
摘要:
采用多聚酶链式反应结合变性高效液相色谱技术建立了发酵乳酸杆菌的快速检测方法。以编码发酵乳酸杆菌Lactobacillus fermentum延伸因子(EF-Tu)基因为靶基因设计引物,优化PCR体系,以加氏乳酸杆菌等26株试验菌株做特异性检测;并将标准菌株稀释成不同梯度,做灵敏度检测。试验结果表明,该方法有很好的特异性,且灵敏度高,检测限可达到100CFU/ml。
关键词:  发酵乳酸杆菌  PCR  变性高效液相色谱  检测
DOI:
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基金项目:国家质检总局课题(2008IK140;2009IK170)资助
Rapid detection of Lactobacillus fermentum by PCR-DHPLC
Abstract:
Rapid detection of Lactobacillus fermentum was established by using polymerase chain reaction (PCR) and denaturing high-performance liquid chromatography (DHPLC). The primers were designed and the PCR system was optimized with the elongation factor (EF-Tu) of L. fermentum as the target gene. Twenty-six strains including L. gasseri were tested with specific detection. The sensitivity of various diluted standard strains were determined. The results indicated that the PCR-DHPLC method was specific and sensitive with a detection limit of 100 CFU/ml.
Key words:  Lactobacillus fermentum  PCR  DHPLC  Detection