二重环介导等温扩增法快速检测乳粉中沙门氏菌和金黄色葡萄球菌

为建立能够同时检测乳粉中沙门氏菌和金黄色葡萄球菌的二重环介导等温扩增(loop-mediated isother-mal amplification,LAMP)方法.针对沙门氏菌invA基因、金黄色葡萄球菌nuc基因保守区域设计LAMP引物,优化引物浓度,并通过熔解曲线分析判断扩增靶标来源,建立同时检测沙门氏菌和金黄色...

Rapid detection of Salmonella and Staphylococcus aureus in milk powder by double loop-mediated isothermal amplification

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.