Tian Xiaolan, Feng Junli, Wang Yi
Institute of Seafood, Zhejiang Gongshang University, Hangzhou 310012, PR China.
Key Lab of aquatic Products Processing of Zhejiang Province, Hangzhou 310012, PR China.
FEMS Microbiol Lett. 2018 Jun 1;365(11). doi: 10.1093/femsle/fny092.
Staphylococcus aureus (S. aureus) is a major human pathogen that may produce a variety of toxins and cause staphylococcal food poisoning. In the present study, a direct loop-mediated isothermal amplification (LAMP) assay was developed and validated to detect S. aureus in food samples. Without prior cultural enrichment and DNA extraction steps, bacterial DNA was released by heating at 100°C for 5 min and directly subjected to LAMP assay. Using a set of LAMP primers recognizing six distinct regions of nuc gene, the developed direct LAMP assay was highly specific, and the analytical sensitivity was determined to be 7.6 × 102 CFU/mL. Moreover, with the pre-mixed LAMP reagents stored at -20°C, the entire assay should be finished within 40 min. These features greatly simplified the operating procedure and made the direct LAMP a powerful tool for rapid and on-site detection of S. aureus in food samples.
金黄色葡萄球菌是一种主要的人类病原体，它可能产生多种毒素并导致葡萄球菌食物中毒。在本研究中，开发并验证了一种直接环介导等温扩增（LAMP）检测方法，用于检测食品样本中的金黄色葡萄球菌。无需事先进行培养富集和DNA提取步骤，通过在100°C加热5分钟释放细菌DNA，然后直接进行LAMP检测。使用一组识别nuc基因六个不同区域的LAMP引物，所开发的直接LAMP检测方法具有高度特异性，分析灵敏度确定为7.6×102 CFU/mL。此外，将预混的LAMP试剂储存在-20°C，整个检测可在40分钟内完成。这些特点极大地简化了操作程序，使直接LAMP成为快速现场检测食品样本中金黄色葡萄球菌有力工具。
