Bacteria Screening, Viability, And Confirmation Assays Using Bacteriophage-Impedimetric/Loop-Mediated Isothermal Amplification Dual-Response Biosensors

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• 作者：Chaker Tlili ; Esen Sokullu ; Mohammadali Safavieh ; Mona Tolba ; Minhaz Uddin Ahmed ; Mohammed Zourob
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：May 21, 2013
• 年：2013
• 卷：85
• 期：10
• 页码：4893-4901
• 全文大小：432K
• 年卷期：v.85,no.10(May 21, 2013)
• ISSN：1520-6882

文摘

Here, we integrate two complementary detection strategies for the identification and quantification of Escherichia coli based on bacteriophage T4 as a natural bioreceptor for living bacteria cells. The first approach involves screening and viability assays, employing bacteriophage as the recognition element in label-free electrochemical impedance spectroscopy. The complementary approach is a confirmation by loop-mediated isothermal amplification (LAMP) to amplify specifically the E. coliTuf gene after lysis of the bound E. coli cells, followed by detection using linear sweep voltammetry. Bacteriphage T4 was cross-linked, in the presence of 1,4-phenylene diisothiocyanate, on a cysteamine-modified gold electrode. The impedimetric biosensor exhibits specific and reproducible detection with sensitivity over the concentration range of 10³鈥?0⁹ cfu/mL, while the linear response of the LAMP approach was determined to be 10²鈥?0⁷ cfu/mL. The limit of detection (LOD) of 8 脳 10² cfu/mL in less than 15 min and 10² cfu/mL within a response time of 40 min were achieved for the impedimetric and LAMP method, respectively. This work provides evidence that integration of the T4-bacteriophage-modified biosensor and LAMP can achieve screening, viability, and confirmation in less than 1 h.