摘要
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。
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 resulTSwere 100%consistent with the resulTSfrom using the traditional methods.The resulTSshow that the new method is stable,rapid and sensitive in identifying E.coli O157:H7 in textiles.
作者
李轲
张子宏
禹建鹰
连素梅
丁友超
谢堂堂
傅科杰
郭会清
LI Ke;ZHANG Zihong;YU Jianying;LIAN Sumei;DING Youchao;XIE Tangtang;FU Kejie;GUO Huiqing(Zhengzhou Customs,Zhengzhou,Henan 450003,China;Shijiazhuang Customs,Shijiazhuang,Hebei 050051,China;Nanjing Customs Industrial ProducTSTesting Center,Nanjing,Jiangsu 210001,China;Shenzhen Customs Industrial ProducTSTesting Technology Center,Shenzhen,Guangdong 518067,China;Ningbo Institute of Inspection Science and Technology,Ningbo,Zhejiang 315000,China)
出处
《纺织学报》
EI
CAS
CSCD
北大核心
2021年第10期92-98,共7页
Journal of Textile Research
基金
河南省科技攻关项目(182102210226)
国家认监委认证认可科技支撑计划项目(2017RJWKJ35)
河南省科技攻关计划项目(162102210370)。
