环介导等温扩增法快速检测阪崎肠杆菌

Rapid Detection of Enterobacter sakazakii by LAMP

目的:研究阪崎肠杆菌的环介导等温扩增(loop-mediated isothermal amplification,简称LAMP)方法,实现对食品中阪崎肠杆菌的快速检测。方法:针对阪崎肠杆菌16S rRNA基因(16S rDNA)及外膜蛋白A(OmpA)基因设计LAMP引物,扩增后通过电泳或加入显色剂SYBR-GREENⅠ检测。结果:LAMP法能有效特异地检测出阪崎肠杆菌;以16S rRNA引物检测阪崎肠杆菌的最低检出限为32CFU/mL,用OmpA引物检测阪崎肠杆菌的最低检出限达3CFU/mL,扩增产物加入SYBR-GREENⅠ在紫外条件下可见荧光,与电泳检测具有同样的灵敏度。结论:LAMP法可实现对阪崎肠杆菌的快速检测,在食品检测中具有广阔的应用前景。

Objective： To develop a loop-mediated isothermal amplification （LAMP） method for rapid detection of Enterobacter sakazakii in food. Methods： LAMP primers were designed on the basis of 16S rRNA gene （16S rDNA） and outer membrane protein A gene （OmpA） in Enterobacter sakazakii. The LAMP products were evaluated by electrophoresis or SYBR-GREEN L Results： The detection limit of Enterobacter sakazakii by LAMP assay was 32 CFU/mL using 16S rRNA primers or 3 CFU/mL using OmpA primers. The similar detection sensitivity of LAMP products using SYBR-GREEN I and electrophoresis was observed. Conclusion： LAMP method was effective and sensitive for rapidly detecting Enterobacter sakazakii, which will have a broad application prospect in food.