Department of Food Science, Cornell University , Stocking Hall, Ithaca, New York 14853, United States.
Department of Food Science, University of Massachusetts , 102 Holdsworth Way, Amherst, Massachusetts 01003, United States.
ACS Sens. 2017 Nov 22;2(11):1573-1577. doi: 10.1021/acssensors.7b00561. Epub 2017 Oct 20.
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 10 CFU mL in food samples without the need for sophisticated instruments or skilled operators.
操作简便、成本低廉且便于运输是专为资源有限环境设计的商用细菌检测试剂盒的关键要求。在这里,我们报告了使用冻干工程噬菌体(噬菌体)对食品样本中的大肠杆菌(E. coli)进行比色检测。在这种方法中,我们对 T7 噬菌体进行了工程改造,使其携带由 T7 启动子驱动的 lacZ 操纵子,以过表达报告酶。工程噬菌体在水溶性聚合物中冻干保存和运输。当用于检测大肠杆菌细胞时,细胞内酶[β-半乳糖苷酶(β-gal)]过表达并释放到周围介质中,提供酶放大的比色信号。使用这种策略,我们能够在无需复杂仪器或熟练操作人员的情况下,在食品样本中检测到浓度为 10 CFU mL 的大肠杆菌细胞。