| 纺织品中大肠杆菌O157:H7的检测方法 |
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| 引用本文： | 李轲,张子宏,禹建鹰,连素梅,丁友超,谢堂堂,傅科杰,郭会清. 纺织品中大肠杆菌O157:H7的检测方法[J]. 纺织学报, 2021, 42(10): 92-98. DOI: 10.13475/j.fzxb.20201101207 |
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| 作者姓名： | 李轲 张子宏 禹建鹰 连素梅 丁友超 谢堂堂 傅科杰 郭会清 |
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| 作者单位： | 1.郑州海关, 河南 郑州 4500032.石家庄海关, 河北 石家庄 0500513.南京海关工业产品检测中心, 江苏 南京 2100014.深圳海关工业品检测技术中心, 广东 深圳 5180675.宁波检验检疫科学技术研究院, 浙江 宁波 315000 |
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| 基金项目： | 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) |
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| 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。
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| 关 键 词： | 纺织品 大肠杆菌O157:H7 毒力基因 免疫磁珠 实时荧光PCR |
| 收稿时间： | 2020-11-26 |
Method to detect virulence gene rfbE of Escherichia coli O157:H7 in textiles |
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| LI Ke,ZHANG Zihong,YU Jianying,LIAN Sumei,DING Youchao,XIE Tangtang,FU Kejie,GUO Huiqing. Method to detect virulence gene rfbE of Escherichia coli O157:H7 in textiles[J]. Journal of Textile Research, 2021, 42(10): 92-98. DOI: 10.13475/j.fzxb.20201101207 |
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| Authors: | LI Ke ZHANG Zihong YU Jianying LIAN Sumei DING Youchao XIE Tangtang FU Kejie GUO Huiqing |
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| Affiliation: | 1. Zhengzhou Customs, Zhengzhou, Henan 450003, China2. Shijiazhuang Customs, Shijiazhuang, Hebei 050051, China3. Nanjing Customs Industrial Products Testing Center, Nanjing, Jiangsu 210001, China4. Shenzhen Customs Industrial Products Testing Technology Center, Shenzhen, Guangdong 518067, China5. Ningbo Institute of Inspection Science and Technology, Ningbo, Zhejiang 315000, China |
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| Abstract: | 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. |
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| Keywords: | textile Escherichia coli O157:H7 virulence gene immunomagnetic bead real-time PCR |
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