Rapid, sensitive, and specific immunomagnetic separation of foodborne pathogens

Emma B. Setterington, Barbara C. Cloutier, J. M. Ochoa, A. Cloutier, Parul Patel, E. Alocilja
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引用次数: 13

Abstract

Food defence requires the means to efficiently screen large volumes of food for microbial pathogens. Even rapid detection methods often require lengthy enrichment steps, making them impractical for this application. There is a great need for rapid, sensitive, specific, and inexpensive methods for extracting and concentrating microbial pathogens from food. In this study, an immunomagnetic separation (IMS) methodology was developed for Escherichia coli O157:H7, using three different types of magnetic nanoparticles (MNPs). The microbiological specificity of the IMS method was evaluated against Escherichia coli O55:H7 and Shigella boydii , and was improved by addition of NaCl during conjugation of antibodies onto MNPs. The microbiological sensitivity of the IMS method was greatest when a high concentration of antibodies (1.0 mg/ml) was present during conjugation. MNP concentrations of 1.0 mg/ml and 0.5 mg/ml provided optimal sensitivity and specificity. The entire IMS procedure requires only 35 minutes, and antibody-conjugated MNPs show no decline in performance up to 60 days after conjugation.
快速、灵敏、特异的食源性致病菌免疫磁分离
食物防御需要有效地筛选大量食物中的微生物病原体。即使是快速检测方法通常也需要冗长的富集步骤,这使得它们不适合这种应用。迫切需要一种快速、灵敏、特异和廉价的方法从食品中提取和浓缩微生物病原体。本研究利用三种不同类型的磁性纳米颗粒(MNPs)建立了大肠杆菌O157:H7的免疫磁分离(IMS)方法。IMS方法对大肠杆菌O55:H7和博氏志贺氏菌的微生物学特异性进行了评估,并通过在MNPs上偶联抗体时添加NaCl提高了其特异性。当偶联过程中存在高浓度抗体(1.0 mg/ml)时,IMS方法的微生物敏感性最高。MNP浓度为1.0 mg/ml和0.5 mg/ml时灵敏度和特异度最佳。整个IMS过程只需要35分钟,抗体偶联的MNPs在偶联后60天内没有表现出性能下降。
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