RAPID DETECTION OF SALMONELLA TYPHIMURIUM USING AN IMMUNOELECTROCHEMICAL METHOD COUPLED WITH IMMUNOMAGNETIC SEPARATION

Journal of Rapid Methods and Automation in Microbiology Pub Date : 2011-11-09 DOI:10.1111/j.1745-4581.1999.tb00371.x

YIHUA CHE, ZHONGPING YANG, YANBIN LI, DAVID PAUL, MICHAEL SLAVIK

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引用次数: 24

Abstract

Abstract An immunoelectrochemical method coupled with magnetic separation was developed for rapid detection of Salmonella typhimurium. Samples containing Salmonella were mixed with anti-Salmonella coated magnetic beads (ASCMB) and alkaline phosphatase labeled anti-Salmonella (APLAS) simultaneously to form ASCMB-Sdhaoaella-APLAS conjugates by antibody-antigen reaction. The conjugates were separated by a magnetic field and were incubated with phenyl phosphate substrate to produce phenol. The number of Salmonella was determined by measuring the phenol concentration using an amperometric tyrosinase biosensor. This method could be completed within two hours with a detection limit of 5 times 10³ cells/mL under optimized conditions (with ***1 mM MgCl₂, 0.2 μg/mL APLAS and ***l mM phenyl phosphate in pH 7.0 TBS). A linear response on semi-log scale was found for Salmonella pure culture between 5 times 10³ and 1 times 10⁶ cells/mL.

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免疫电化学联用免疫磁分离快速检测鼠伤寒沙门菌

摘要建立了磁分离-免疫电化学快速检测鼠伤寒沙门菌的方法。将含沙门菌样品与抗沙门菌包被磁珠(ASCMB)和碱性磷酸酶标记的抗沙门菌(APLAS)同时混合，通过抗体-抗原反应形成ASCMB- shaoaella -APLAS偶联物。用磁场分离共轭物，并与磷酸苯酯底物孵育产生苯酚。采用电流型酪氨酸酶生物传感器测定苯酚浓度，确定沙门氏菌数量。在优化条件下(***1 mM MgCl2, 0.2 μg/mL APLAS， ***1 mM苯基磷酸，pH 7.0 TBS)， 2 h内检出限为5 × 103 cells/mL。沙门氏菌纯培养在5 × 103 ~ 1 × 106细胞/mL之间呈半对数线性关系。

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Journal of Rapid Methods and Automation in Microbiology 生物-生物工程与应用微生物

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