Rapid detection of Salmonella and Staphylococcus aureus in milk powder by double loop-mediated isothermal amplification
XU Wenwen1, LIANG Yulin1, WANG Yunxia2, LIU Xiu1*, YIN Jianjun1, ZHOU Guangjun3, SONG Quanhou1, DING Mengxuan1, ZHOU Pengfei1
1(China National Research Institute of Food & Fermentation industries Co.,Ltd.,Beijing 100015,China) 2(Inner Mongolia Yili Industrial Group Limited by Share Ltd.,Hohhot 010110,China) 3(Shandong daoxiangcun Food Industry Co.,Ltd.,Heze 274100,China)
Abstract: The aim of this research is to establish a loop-mediated isothermal amplification (LAMP) method that can simultaneously detect Salmonella and Staphylococcus aureus in milk powder.LAMP primers were designed based on the conserved region of invA gene in Salmonella and the nuc gene in S.aureus.After primer concentration optimization and melting curve analysis to determine the source of amplification target,a double LAMP detection method for simultaneous detection of Salmonella and S.aureus was established.The results showed that when the primer concentration ratio of Salmonella to S.aureus was 3∶1,the established double detection system had the best amplification efficiency.The detection system had good specificity and high sensitivity.Six target strains and nine non-target strains were tested,resulting in no false positive and false negative result.The sensitivity of the two foodborne pathogens reached 102 fg/μL.In addition,according to the annealing temperature in the melting curve,the target strain can be accurately judged to complete the strain differentiation.The established detection method can be used for rapid on-site screening of Salmonella and S.aureus in milk powder samples of dairy companies.
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2021, Vol. 47 
