现代食品科技2024，Vol.40Issue(4)：287-295,9.DOI:10.13982/j.mfst.1673-9078.2024.4.0230
恒温隔绝式PCR快速检测蜡样芽孢杆菌方法的建立及应用
Development of an Insulated Isothermal PCR-based Method for Rapid Detection of Bacillus cereus
摘要
Abstract
Specific primers and probe were designed for the gyr B gene of Bacillus cereus encoding the gyrase B subunit,and a method based on insulated isothermal polymerase chain reaction(iiPCR)was established for the rapid detection of B.cereus and subsequently validated.The efficacy of this method was compared with that of traditional PCR and real-time fluorescent quantitative PCR(SYBER Green Ⅰ fluorescent dye and TaqMan probe methods)detection methods,and applied in the detection of B.cereus in different types of retail food.The results revealed that the newly developed method facilitated the rapid detection of B.cereus within 40 min.Moreover,there was no cross reaction with other bacteria,including Escherichia coli,Staphylococcus aureus,Listeria monocytogenes,and Salmonella enteritidis,thereby indicating good specificity.The minimum detection limit of the method was 1.5×102 CFU/mL,which was significantly superior to that of the traditional PCR method,and equivalent to that of quantitative PCR.Of the 42 retail food samples assessed,19(45.23％)were found to be contaminated with B.cereus.The results obtained for method validated were completely consistent with those obtained using traditional PCR detection,although test results were available within a considerably shorter timeframe,saving at least 4 h.The method developed in this study provides a rapid,convenient,specific,and sensitive procedure that is suitable for the real-time on-site detection of B.cereus.关键词
蜡样芽孢杆菌/恒温隔绝式PCR/gyrB基因Key words
Bacillus cereus/insulated isothermal PCR/gyrB gene引用本文复制引用
寇肖迪,孟含,苏雅航,汤承,胡瑜,唐俊妮..恒温隔绝式PCR快速检测蜡样芽孢杆菌方法的建立及应用[J].现代食品科技,2024,40(4):287-295,9.基金项目
国家重点研发计划重点专项(2018YFD0500500) （2018YFD0500500）
西南民族大学研究生创新型科研项目(ZD2022257) （ZD2022257）
