Rapid detection of coliphage

The need for rapid detection Of coliphage is something that has been present for years. Coliphage is fecal-borne viruses. These can be present in virtually any body of water in any country in the world. The present method of coliphage detection takes at least 24 hours once the sample arrives at the lab. It requires a lab setting with delicate equipment and highly trained personnel. We are striving to create a handheld device that takes 1-3 hours and can be performed by anyone.

We plan to use freeze-dried E. coli cells in a hand held device. The [potentially] contaminated water will be introduced in controlled volumes and lyse (burst) the cells. The amount of virus present will be indicated with a colorimetric reading.

We have constructed a plasmid which we have transformed into a strain of Escherichia coli. This plasmid consists of a promoter, a mutant lacZ operon, and a terminator. The enzyme fragment produced by the lacZ operon is released upon lysis and seeks out its complement in vitro (outside of the cell.)The enzymatic activity of this complementation reacts with our indicator, chlorophenol red (CPRG) to produce a color change that is directly proportional to the amount of viruses present.