摘要
建立针对β型溶血性链球菌的变性泡介导恒温扩增检测方法,对检测方法进行完整的方法学评价试验。对检测方法的反应温度进行优化得到最适温度为60℃,在此基础上,以常见菌株特异性测试、稀释模板灵敏度测试及10组乳粉样本稳定性测试3方面对方法进行方法学评价,并对特异性测试进行荧光可视化检测。方法学评价结果显示所建立的β型溶血性链球菌的变性泡介导恒温扩增检测方法的最快检测时间可达到30 min以内,5种常见的食源性致病菌特异性试验结果无误,灵敏度试验得到的荧光值趋势良好,100 ng/μL条件下最低荧光阈值时间最快,可达到30 min以内,3位实验员对该方法与国家标准方法的对比结果未出现异常和不符,可视化检测结果与荧光监测结果一致。该方法特异性、灵敏性、稳定性均良好。
To establish a denatured vesicle-mediated constant temperature amplification detection method for betahemolytic Streptococcus,and conduct a complete methodological evaluation on the detection method.the reaction temperature of the detection method was optimized and the optimal temperature was 60℃.On this basis,the detection method was evaluated from three aspects:common strain specific testing,dilution template sensitivity testing and stability testing of 10 groups of milk powder sample.Fluorescence visualization detection was performed on the specificity testing.The results of the methodological evaluation showed that the fastest detection time of the denatured vesicle-mediated constant temperature amplification detection method for beta-hemolytic Streptococcus could reach within 30 minutes,the specific test results of five common foodborne pathogenic bacteria were correct,and the fluorescence values obtained by the sensitivity test showed a good trend,the lowest fluorescence threshold time was the fastest at 100ng/μL concentration,which could reach within 30min.The results of the comparison between the method and the national standard method by 3 experimenters showed no abnormalities or discrepancies,and the visual detection results were consistent with the fluorescence monitoring results.The method had good specificity,sensitivity and stability.
作者
史贇学
杨春梅
吴雪莉
冉凌云
李敏婕
邓玲
孙家欣
姚华
SHI Yunxue;YANG Chunmei;WU Xueli;RAN Lingyun;LI Minjie;DENG Ling;SUN Jiaxin;YAO Hua(Chengdu Institute of Product Quality Inspection Co.,Ltd.,Chengdu 610000)
出处
《中国食品添加剂》
CAS
2024年第4期243-248,共6页
China Food Additives
基金
四川省科技厅项目(2023YFN0059)
国家市场监督管理总局科技计划项目(2021MK096)。
关键词
变性泡介导
恒温扩增
溶血性链球菌
方法学评价
denatured vesicle-mediated
constant temperature amplification
hemolytic Streptococcus
methodological evaluation
作者简介
史贇学(1993-),男,硕士,助理工程师,研究方向:食品安全与控制方向。E-mail:872551416@qq.com;通信作者:姚华(1985-),男,本科,工程师,研究方向:食品微生物检测。E-mail:460699297@qq.com。
