摘要
mecA是耐甲氧西林葡萄球菌携带的耐药基因。本实验根据克隆的mecA基因序列,在保守区设计了一套环介导等温扩增(LAMP)引物,通过条件优化,建立了耐甲氧青霉素mecA基因LAMP检测方法,其最佳工作条件为:甜菜碱浓度为0.8 mol/L,Mg2+浓度为20 mmol/L,反应温度63℃,时间50 min。此方法可在60 min左右完成对mecA基因的检测,特异性好,mecA基因的LAMP检测灵敏度比PCR高100倍。对38株葡萄球菌分离株的mecA基因的检测表明,阳性携带率为44.7%。
mecA is unique as a resistance gene of methicillin-resistant Staphylococccus aureus（MRSA）.Based on the cloned mecA gene in the assay,a set of primers was designed against highly conserved sequences respectively,then the LAMP reaction conditions were optimised and the detection for the mecA gene using loop-mediated isothermal amplication were established initially.The final conditions were fixed,betaine 0.8 mol/L,Mg2＋ 20 mmol/L,completed within 50 min in 63 ℃ isothermal water-bath.Compared to PCR detection,this detection can be completed in 60 min with good specificity and more rapidity,100 times higher sensitivity for the mecA gene.The positive rate of the mecA gene is 44.7 % in 38 staphylococcus aureus isolated.
出处
《食品研究与开发》
CAS
北大核心
2011年第8期68-71,共4页
Food Research and Development
基金
国家质检总局科技计划项目(2009GFW-0251)
