Colorimetric detection of Escherichia coli using engineered bacteriophage and an affinity reporter system.

ABSTRACT: Reporter phage systems have emerged as a promising technology for the detection of bacteria in foods and water. However, the sensitivity of these assays is often limited by the concentration of the expressed reporter as well as matrix interferences associated with the sample. In this study, bacteriophage T7 was engineered to overexpress mutated alkaline phosphatase fused to a carbohydrate-binding module (ALP*-CBM) following infection of E. coli to enable colorimetric detection in a model system. Magnetic cellulose particles were employed to separate and concentrate the overexpressed ALP*-CBM in bacterial lysate. Infection of E. coli with the engineered phage resulted in a limit of quantitation of 1.2 × 10⁵ CFU, equating to 1.2 × 10³ CFU/mL in 3.5 h when using a colori

SUBMITTER: Singh S 

PROVIDER: S-EPMC7241434 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature