基于多壁碳纳米管/SDBS/硫堇阪崎肠杆菌免疫传感器的研究  

Studies on Immunosensor for Enterobacter Sakazakii Based on Multi-Wall Carbon Nanotubes/SDBS and Thionine

作　　者：张晓[1] 窦文超[1] 赵广英[1] 

机构地区：[1]浙江工商大学食品与生物工程学院浙江省食品安全重点实验室,杭州310035

出　　处：《中国食品学报》2012年第2期124-130,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金资助项目(No.30571623);浙江省重中之重项目(No.Z05-178)

摘　　要：目的:研制一种灵敏度高、电化学性能稳定的免疫传感器,构建准确检测阪崎肠杆菌(Enterobacter sakazakii,E.sakazakii)的电化学方法。方法:将硫堇(Thi)、辣根过氧化物酶标记的阪崎肠杆菌抗体(HRP-anti-E.sakazakii)依次自组装固定于多壁碳纳米管(MWCNT)/十二烷基苯磺酸钠(SDBS)修饰的四通道丝网印刷电极上,制得一次性免疫传感器。采用原子力显微镜(AFM)表征电极修饰和孵育抗原后的表面形态,用循环伏安法(CV)考察不同修饰电极的电化学特性,根据还原峰电流的变化对阪崎肠杆菌进行测定。结果:在优化的实验条件下,该方法检测阪崎肠杆菌的线性范围为102～108cfu/mL,检测限为5.7×101cfu/mL(S/N=3)。结论:该免疫传感器灵敏度很高,具有较好的特异性、重现性(RSD=6.3%)、稳定性(4℃无菌容器中放置15d后电流响应为初始值的93.24%)和准确性(与GB/T4789.40-2010符合率为96.67%),具有一定的应用潜力。Object： For the purpose of establishing the accurate electrochemical method for detecting Enterobacter sakazkii （E. sakazakii）, a sensitive and stable immunosensor was successfully fabricated. Method： First, multi-wall carbon nanotube （MWCNT）/sodium dodecylbenzene sulfonate （SDBS） homogeneous composite was dropped on the surface of the 4-channel screen-printed carbon electrode. Then Thionine was chemisorbed by carboxylic MWCNT. Furthermore, horseradish peroxidase labeled antibody to E. sakazakii （HRP-anti-E.sakazakii） was chemisorbed onto Thionine filmthrough the electrostatic force with the amino group of Thionine. The preparation process of modified electrode was char- acterized with atomic force microscope （AFM）. Cyclic voltammetry （CV） was carried out to characterize the electrochemi- cal properties of different electrodes and detect E. sakazakii. Results： under optimal conditions, concentration of E. sakazakii from 102-10Scfu/mL could be detected with a detection limit of 5.7x10~ cfu/mL （S/N=3）. Conclusions： The en- zyme immunosensor exhibited high specificity, satisfactory reproducibility （RSD=6.3%）, stability （electrical response was 93.24% of initial value after storage in sterile container at 4℃ for 15 d） and accuracy（coincidence rate was 96.67% with GB/T 4789.40-2010）. The novel had potential application for rapid detection

关 键 词：免疫传感器 多壁碳纳米管 硫堇 阪崎肠杆菌 快速检测 

分 类 号：TS207.4[轻工技术与工程—食品科学] TP212.2[轻工技术与工程—食品科学与工程]