| 侧向流动型重组酶聚合酶扩增技术检测食品中大肠埃希氏菌O157 |
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| 引用本文： | 兰全学,陈佳平,杨慧,甘莉萍,杜文旗,刘渠,金玉娟.侧向流动型重组酶聚合酶扩增技术检测食品中大肠埃希氏菌O157[J].食品科技,2021(1). |
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| 作者姓名： | 兰全学 陈佳平 杨慧 甘莉萍 杜文旗 刘渠 金玉娟 |
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| 作者单位： | 深圳市龙岗区疾病预防控制中心;深圳市计量质量检测研究院 |
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| 基金项目： | 深圳市科创委项目(JSGG20160606144217004)。 |
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| 摘 要： | 研究内容基于侧向流动型重组酶聚合酶扩增(Lateral flow-based recombinase polymerase amplification,LF-RPA)技术原理,建立大肠埃希氏菌O157(Escherichia coli O157)的LF-RPA快速检测方法。应用该方法对3株大肠埃希氏菌O157和27株非大肠埃希氏菌O157进行检测,结果表明该方法具有良好的特异性。分别以10倍浓度梯度稀释插入靶基因序列的质粒pUC57-rfbE和大肠埃希氏菌O157菌液DNA作为模板进行LF-RPA检测。结果表明LF-RPA方法可检出1.0×102拷贝/μL的pUC57-rfbE质粒和1.2×104 cfu/mL的大肠埃希氏菌O157菌液。以大肠埃希氏菌O157污染食品样品,增菌4 h后,大肠埃希氏菌O157初始浓度为1.2×101 cfu/mL的食品增菌液可以被LF-RPA检出阳性信号。该研究建立的方法可快速、准确、灵敏地检测食品中大肠埃希氏菌O157,无需专业设备,在食品生产企业和基层检测机构具有一定的应用前景。
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| 关 键 词： | 重组酶聚合酶扩增 侧向流动免疫分析 大肠埃希氏菌O157 rfbE基因 |
The Detection Method for Escherichia Coli O157 Based on Lateral Flow-Based Recombinase Polymerase Amplification |
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| LAN Quanxue,CHEN Jiaping,YANG Hui,GAN Liping,DU Wenqi,LIU Qu,JIN Yujuan.The Detection Method for Escherichia Coli O157 Based on Lateral Flow-Based Recombinase Polymerase Amplification[J].Food Science and Technology,2021(1). |
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| Authors: | LAN Quanxue CHEN Jiaping YANG Hui GAN Liping DU Wenqi LIU Qu JIN Yujuan |
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| Affiliation: | (Shenzhen Longgang District Center for Disease Control and Prevention,Shenzhen 518172,China;Shenzhen Academy of Metrology&Quality Inspection,Shenzhen 518109,China) |
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| Abstract: | This study developed a rapid detection method for Escherichia coli O157 based on lateral flow-based recombinase polymerase amplification (LF-RPA).According to testing 3 Escherichia coli O157 and 27 non-Escherichia coli O157 strains by using LF-RPA,the developed method in this research can detect Escherichia coli O157 specifically.To carry out LF-RPA assay with the templates of tenfold gradient diluted pUC57-rfbE plasmids;moreover,LF-RPA assay was carried out by using the templates of extracted DNA from tenfold gradient diluted Escherichia coli O157 liquid.The sensitivity of LF-RPA can reach 1.0×102 copies/μL and 1.2×104 cfu/mL,respectively.The food samples were inoculated with Escherichia coli O157,positive signals can be obtained from the food samples inoculated with 1.2×101 cfu/mL Escherichia coli O157 after 4 h enrichment.This study developed a testing method for Escherichia coli O157,this method can detect Escherichia coli O157 rapidly,specifically and sensitively without professional equipment.So Escherichia coli O157 LF-RPA can be used for point-of-care detection of Escherichia coli O157 in food production enterprises and grass-roots testing institutions. |
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| Keywords: | recombinase polymerase amplification lateral flow immunoassay Escherichia coli O157 rfbE gene |
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