牛奶中β-内酰胺类抗生素残留检测试剂盒的研制.pdfVIP

现代食品科技 Modern Food Science and Technology 2014, Vol.30, No.10 牛奶中β-内酰胺类抗生素残留检测试剂盒的研制 刘冬梅，郭均，梁洁英，吴晖，余以刚，李晓凤，唐语谦 （华南理工大学轻工与食品学院，广东广州 510640） 摘要：以牛奶中残留的β-内酰胺类抗生素为检测目标，开发一种能快速检测其残留的试剂盒。利用嗜热脂肪芽孢杆菌BS 10208 对酸和β-内酰胺类抗生素敏感的特点，优化其最优的生长条件，并制成在2.5 h 内能发生显色反应的检测试剂盒。单因素结果表明， 当菌液接种量5%、生长初始pH 8.0、培养温度55 ℃、蛋白胨添加量0.50%，其菌落数对数值分别为2.9、7.5、7.3、5.8。正交结果 表明，在外加入0.5%蛋白胨营养肉汤中，接种5%种子液，在65 ℃恒温振荡器中培养24 h ，其菌落数对数值为7.6 。优化后检测试 剂盒在2.5 h 内能检测牛奶中-内酰胺类抗生素残留，其各项参数为：反应体积为0.5 mL，1.5%琼脂，0.01 g/L 溴甲酚紫浓度，BS 10208 细胞总数为1.30×108 cfu，最适反应温度65 ℃，青霉素钠的限值为24 g/L。该试剂盒还能检测牛奶中残留的四环素类、磺胺类、 大环内酯类、氨基糖苷类等抗生素。 关键词：嗜热脂肪芽孢杆菌；β- 内酰胺类抗生素；牛奶；溴甲酚紫 文章篇号：1673-9078(2014)10-251-256 DOI: 10.13982/j.mfst.1673-9078.2014.10.042 Development of a Test Kit forβ-lactam Antibiotic Residues in Milk LIU Dong-mei, GUO Jun, LIANG Jie-ying, WU Hui, YU Yi-gang, LI Xiao-feng, TANGYu-qian (College of Light Industry and Food Science, South China University of Technology, Guangzhou 510640, China) Abstract: We used β-lactam antibiotic residues in milk as the target to develop a rapid detection kit. Based on sensitivity of Bacillus stearothermophilus 10208 to acid and β-lactam antibiotics, growth conditions were optimized and a test kit that could show a color reaction within 2.5 h was developed. The results of the single-factor experiment showed that when the inoculum was 5%, initial growth pH was 8.0, culture temperature was 55 ℃, and the quantity of peptone was 0.50%, the logarithm values of growth for the bacterium were 2.9, 7.5, 7.3, and 5.8, respectively. The results of an orthogonal test showed that when a nutrient broth containing 0.50% peptone was added and the mixture was inoculated with 5% seed broth and incubated for 24 h at 65 ℃, the logarithm value of growth for the bacterium was 7.6. T