摘要
为了建立原料乳中金黄色葡萄球菌的环介导等温扩增(LAMP)检测方法,本研究根据金黄色葡萄球菌的耐热核酸酶基因(nuc)在保守区域设计特异性引物,建立LAMP检测方法,并用于原料乳中金黄色葡萄球菌的快速检测。结果表明,针对金黄色葡萄球菌的基因组DNA,使用设计合成的4条引物和建立的LAMP方法能够有效检测金黄色葡萄球菌DNA,每25.0μl反应体系的灵敏度为110.0 fg,并且此方法针对金黄色葡萄球菌具有良好的特异性。针对原料乳中金黄色葡萄球菌的污染,建立的LAMP检测方法的检测限为67 CFU/ml。本研究结果为进一步将LAMP方法应用于食品中金黄色葡萄球菌的安全检测奠定了基础。
The loop-mediated isother^nal amplification(LAMP) method was established and applied in the rapid de-tection of Staphylococcus aureus in raw milk according to the conser^^ed sequence of the heat-tolerant nuclease gene ( n u c ) . The genomic DNA of S. aureus could be detected using the four primers designed and synthesized,and the detection limit was 110. 0 fg per 25. 0μl reaction system. The method was S. aureus specific,with the detection limit of 67 CFU/ml in raw milk. This study provided a basis for further application of LAMP method in the safety detection of S. aureus in food.
出处
《江苏农业学报》
CSCD
北大核心
2017年第5期1171-1175,共5页
Jiangsu Journal of Agricultural Sciences
基金
北京市长城学者培养计划项目(CIT&TCD20140315)
北京农学院研究生改革与发展项目(2016YJS048)