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Lyophilized Engineered Phages for Escherichia coli Detection in Food Matrices
ACS Sensors ( IF 8.2 ) Pub Date : 2017-10-20 00:00:00 , DOI: 10.1021/acssensors.7b00561 Juhong Chen 1 , Rachael A. Picard 2 , Danhui Wang 1 , Sam R. Nugen 1
ACS Sensors ( IF 8.2 ) Pub Date : 2017-10-20 00:00:00 , DOI: 10.1021/acssensors.7b00561 Juhong Chen 1 , Rachael A. Picard 2 , Danhui Wang 1 , Sam R. Nugen 1
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Ease of use, low cost, and convenient transport are the key requirements for a commercial bacteria detection kit designed for resource-limited settings. Here, we report the colorimetric detection of Escherichia coli (E. coli) in food samples using freeze-dried engineered bacteriophages (phages). In this approach, we have engineered T7 phages to carry the lacZ operon driven by T7 promoter to overexpress reporter enzymes. The engineered phages were freeze-dried in a water-soluble polymer for storage and transportation. When used for the detection of E. coli cells, the intracellular enzyme [β-galactosidase (β-gal)] was overexpressed and released into the surrounding media, providing an enzyme-amplified colorimetric signal. Using this strategy, we were able to detect E. coli cells at the concentration of 102 CFU mL–1 in food samples without the need for sophisticated instruments or skilled operators.
中文翻译：
用于食品基质中大肠杆菌检测的冻干工程噬菌体
易于使用，成本低廉，运输方便是专为资源有限的环境设计的商用细菌检测套件的关键要求。在这里，我们报告使用冷冻干燥的工程噬菌体（噬菌体）对食品样品中的大肠杆菌（E. coli）进行比色检测。在这种方法中，我们设计了T7噬菌体以携带由T7启动子驱动的lacZ操纵子来过表达报告酶。将工程化的噬菌体在水溶性聚合物中冷冻干燥，以进行存储和运输。当用于大肠杆菌的检测时在细胞中，细胞内酶[β-半乳糖苷酶（β-gal）]被过表达并释放到周围的培养基中，从而提供了一种酶放大的比色信号。使用这种策略，我们无需复杂的仪器或熟练的操作人员就能检测食物样品中10 2 CFU mL –1浓度的大肠杆菌细胞。
更新日期：2017-10-20
中文翻译：
用于食品基质中大肠杆菌检测的冻干工程噬菌体
易于使用，成本低廉，运输方便是专为资源有限的环境设计的商用细菌检测套件的关键要求。在这里，我们报告使用冷冻干燥的工程噬菌体（噬菌体）对食品样品中的大肠杆菌（E. coli）进行比色检测。在这种方法中，我们设计了T7噬菌体以携带由T7启动子驱动的lacZ操纵子来过表达报告酶。将工程化的噬菌体在水溶性聚合物中冷冻干燥，以进行存储和运输。当用于大肠杆菌的检测时在细胞中，细胞内酶[β-半乳糖苷酶（β-gal）]被过表达并释放到周围的培养基中，从而提供了一种酶放大的比色信号。使用这种策略，我们无需复杂的仪器或熟练的操作人员就能检测食物样品中10 2 CFU mL –1浓度的大肠杆菌细胞。
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