A new replication medium enables a rapid identification of {varphi}x-174 virus by synchronous fluorescence of Tryptophan.

Abstract

Development of rapid methods for detection of virus particles based on their intrinsic fluorescence is challenging. Pure viruses may be detected in filtered solutions, based on the strong fluorescence of the amino acid Tryptophan (Trp) in their proteins. Nevertheless, Trp also exists in high quantities in the hosts and host cultivation media. In this work, we show that a separation of the bacteriophage{varphi} x-174 from its E. coli host (grown on the standard cultivation medium "Nutrient Agar") by simple extraction and filtration is not sufficient for its detection based on the intrinsic fluorescence since ~70 % of the Trp fluorescence is derived from impurities. We formulate a new cultivation medium with very low Trp concentration. We apply synchronous fluorescence measurements to show that no Trp fluorescence is detected in the extract solution upon incubation of this medium substrate with ammonium acetate extraction buffer. Finally, we detect{varphi} x-174 based on the spectral fingerprint of its intrinsic Trp content by synchronous fluorescence measurements. The concept of coupling intrinsic fluorescence-based methods to impurities reduction in the source, may pave the way towards future development of simple, cheap, and rapid methods for detection of viral pathogens.