纺织品中大肠杆菌O157:H7的检测方法

为建立一种灵敏、特异、快速、高效地鉴定纺织品基质中大肠杆菌O157:H7的方法,筛选出大肠杆菌O157:H7特有的rfbE基因保守序列,设计PCR特异性引物和荧光双标记探针,结合免疫磁珠技术,集成创新开发一种目标菌低浓度、不可培养情况下的样品中高效、快速富集分离大肠杆菌O157:H7的技术,建立了一种针对纺织品基质的免疫磁珠富集-实时荧光定量PCR方法,其检测下限可达8 CFU/mL。利用该方法对收集到的30份阳性样品进行鉴定,鉴定结果与传统方法结果100%吻合。结果表明,新建方法稳定性强,实现了纺织品中大肠杆菌O157:H7的快速灵敏鉴定。

Method to detect virulence gene rfbE of Escherichia coli O157:H7 in textiles

In order to establish a sensitive, specific, rapid and efficient method to evaluate E. coli O157:H7 in textiles, the conservative rfb E sequence of E. coli O157:H7 was screened for the design of PCR specific primers and fluorescent double labeled probes. Combined with immunomagnetic beads technology, a new technology was developed for quick and efficient isolation of E. coli O157:H7 under the condition of low concentration and non-culturable target bacteria. A real-time fluorescent quantitative PCR method was developed, and the detection limit could reach 8 CFU/g. 30 positive samples were identified using this method, and the results were 100% consistent with the results from using the traditional methods. The results show that the new method is stable, rapid and sensitive in identifying E. coli O157:H7 in textiles.