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杨大伟1,2， 刘云国2*， 雷质文2， 周裔斌1， 陈俊芳1， 王建广2
（1. 安徽农业大学 茶与食品科技学院，安徽 合肥 230000;
2. 山东出入境检验检疫局 检验检疫技术中心，山东 青岛 266002）
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凝结芽孢杆菌的快速检测方法。以编码凝结芽孢杆菌的特异性基因寡聚-1,6-葡萄糖苷酶基因作为靶基因的设计引物，优化PCR体系，以嗜热脂肪芽孢杆菌等23株试验菌株做特异性检测。同时，将标准菌株稀释成不同梯度，做灵敏度检测。与传统的检测方法相比，该方法有很好的特异性，且灵敏度高，检测时间短并可节省人力，具有快速方便等优点，检测限可达到100 cfu·mL-1，适用于食物中凝结芽孢杆菌的快速检测。
关键词: 凝结芽孢杆菌； PCR； 变性高效液相色谱； 检测
中图分类号: O 652.63 文献标志码： A
Denaturing High Performance Liquid Chromatography Used in the Detection of Bacillus Coagulans
YANG Da-wei1，2, LIU Yun-guo2, LEI Zhi-wen2, ZHOU Yi-bin1, CHEN Jun-fang1, WANG Jian-guang2
(1. College of Tea and Food Science and Technology, Anhui Agriculture University,Hefei 230000,China;
2. Inspection and Qusrantine Technology Center, Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002,China)
Abstract: In this study, we reported a new method of PCR-DHPLC to detect Bacillus coagulans. By using gene of Bacillus coagulans’ oligomerization-1,6-glucosidase as the target gene, PCR system was optimized and the twenty-three test strains from Bacillus stearothermophilus were tested by specific detection. Then different grades of standard strains were obtained by diluting and sensitivity was detected. Compared with the conventional method. The PCR-DHPLC methods could test Bacillus coagulans exclusively and sensitively with the lowest amount of 100 cfu·mL-1. The PCR-DHPLC method was rapid and accurate in detection of Bacillus coagulans in food.
Key words: Bacillus coagulans； PCR； DHPLC； detection
收稿日期：2010-11-03
基金项目： 国家质检总局课题项目（2008IK140；2009IK170）.
作者简介： 杨大伟（1984—），男，硕士研究生. * 通信联系人.
