PCR couple with magnetic nanoparticles-based biotin-avidin system for chemiluminescent detection of specific gene (rfbE) of E. coli O157

摘要：

A novel method was established through the amplification of chemiluminescent signals of nucleic acid hybridization based on magnetic nanoparticles (MNPs) and biotin-avidin system. Amino-modified specific probe was firstly coupled to MNPs with carboxylic acid. Then the probes on the MNPs were hybridized with the biotin-dUTP-labeled DNA fragments obtained by PCR amplification. Streptavidin-modified alkaline phosphatase was applied to bind with the biotin, and the alkaline phosphatase reacted with 3-(2-spiroadamantane)-4-methoxy-4-(3-phosphoryloxy) phenyl-1, 2-dioxetane to generate a light-emitting product. Specific gene (rfbE) of E. coli O157 antigen was studied to evaluate this system. The MNPs immobilized with specific probe of rfbE could specifically capture the biotin-labeled DNA fragment of rfbE rather than that of the chloramphenicol acetyltrans-ferase (CAT) gene.

关键词： magnetic nanoparticles PCR chemiluminescence alkaline phosphatase

作者: Li Zhi-Yang He Lei He Nong-Yue Li Song Liu Hong-Na

作者单位: The State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China School of Public Health, Southeast University, Nanjing, 210009, China

会议类型: 国际会议

会议名称: International Symposium on Medical and Pharmaceutical Biotechnology(医药生物技术国际研讨会)

会议地点: 南京

会议语种:英文

页码: 123-125

在线出版日期: 2009-04-27（万方平台首次上网日期，不代表论文的发表时间）