Chemically immobilized T4-bacteriophage for specific Escherichia coli detection using surface plasmon resonance
Chemically immobilized T4-bacteriophage for specific Escherichia coli detection using surface plasmon resonance
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DOI：
10.1039/c0an00697a
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发表时间：
2011-01-01
期刊：
影响因子：
4.2
通讯作者：
Evoy, S.
中科院分区：
文献类型：
--
作者：
Arya, Sunil K.;Singh, Amit;Evoy, S.
建立了以T4噬菌体(T4)为特异性受体，以表面等离子体共振(SPR)为转导技术的检测大肠杆菌K12细菌的生物检测平台。T4噬菌体通过自组装的丁二胺基丙酸酯(DTSP)单分子膜固定在金表面。用扫描电子显微镜(SEM)对不同T4噬菌体浓度制备的BSA/EA-T4/DTSP/Au底物进行了表征。研究表明，DTSP的使用导致T4噬菌体均匀结合在表面。SPR分析表明，这些BSA/EA-T4/DTSP/Au接口可以检测到对非宿主大肠杆菌NP10和NP30具有高特异性的大肠杆菌K12。扫描电子显微镜和SPR研究结果表明，当T4噬菌体的固定化浓度为1.5×10(11)pfu ml(-1)时，宿主细菌捕获率最高。这些化学锚定的噬菌体底物的表面可以再生，用于重复检测E.ColiK12，并可用于7×10(2)至7×10(8)cfu ml(-1)的检测。这些研究的结果对利用噬菌体识别固有的选择性开发在线生物检测方法以检测各种食物和水传播的病原体具有重要意义。
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 x 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 x 10(2) to 7 x 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.