Department of Electrical and Computer Engineering, University of Alberta, Edmonton, Canada.
Analyst. 2011 Feb 7;136(3):486-92. doi: 10.1039/c0an00697a. Epub 2010 Nov 16.
A bioassay platform using T4 bacteriophage (T4) as the specific receptor and surface plasmon resonance (SPR) as the transduction technique has been developed for the detection of Escherichia coli K12 bacteria. The T4 phages have been covalently immobilized onto gold surfaces using a self-assembled monolayer of dithiobis(succinimidyl propionate) (DTSP). Substrates of BSA/EA-T4/DTSP/Au prepared using different T4 phage concentrations have been characterized using scanning electron microscopy (SEM). The studies reveal that the use of DTSP results in a uniform binding of T4 phages onto the surface. The SPR analysis demonstrates that these BSA/EA-T4/DTSP/Au interfaces can detect the E. coli K12 with high specificity against non-host E. coli NP10 and NP30. Results of SEM and SPR studies indicate that the maximum host bacterial capture is obtained when 1.5 × 10(11) pfu ml(-1) concentration of T4 phages was used for immobilization. The surface of these chemically anchored phage substrates can be regenerated for repeated detection of E. coli K12 and can be used for detection in 7 × 10(2) to 7 × 10(8) cfu ml(-1) range. The results of these studies have implications for the development of online bioassays for the detection of various food and water borne pathogens using the inherent selectivity of bacteriophage recognition.
已开发出一种使用 T4 噬菌体（T4）作为特异性受体和表面等离子体共振（SPR）作为转导技术的生物测定平台，用于检测大肠杆菌 K12 细菌。T4 噬菌体已使用二硫代双（琥珀酰亚胺基丙酸酯）（DTSP）的自组装单层共价固定在金表面上。使用不同 T4 噬菌体浓度制备的 BSA/EA-T4/DTSP/Au 底物已使用扫描电子显微镜（SEM）进行了表征。研究表明，DTSP 的使用导致 T4 噬菌体在表面上均匀结合。SPR 分析表明，这些 BSA/EA-T4/DTSP/Au 界面可以高度特异性地检测到大肠杆菌 K12，而对非宿主大肠杆菌 NP10 和 NP30 则具有特异性。SEM 和 SPR 研究的结果表明，当使用 1.5×10(11)pfu ml(-1)浓度的 T4 噬菌体进行固定时，可获得最大的宿主细菌捕获量。这些化学锚定噬菌体底物的表面可以进行再生，以重复检测大肠杆菌 K12，并可用于检测 7×10(2)至 7×10(8)cfu ml(-1)的范围内。这些研究的结果对于使用噬菌体识别的固有选择性开发用于检测各种食源和水源病原体的在线生物测定具有重要意义。
