Integrated Cell Culture-Mass Spectrometry Method for Infectious Human Virus Monitoring

Tools for detecting infectious human viruses in environmental samples are essential for protecting human health. Here, we developed an Integrated Cell Culture-Mass Spectrometry (ICC-MS) method that detects infectious viruses by identifying viral proteins that replicate in cell cultures. The ICC-MS method was first validated and optimized with murine hepatitis viruses (MHV) spiked in culture media and wastewater concentrates and then applied to detect human viruses in untreated and treated wastewater samples. Reoviruses were detected in wastewater samples when the method was applied with Vero, BGMK, and BSC-1 cell lines. Strain-specific viral peptides from reovirus type 1, type 2, and type 3 were detected in primary influent, secondary effluent (pre-UV treatment), and UV-treated effluent. We propose that the ICC-MS method is a promising technique for directly detecting infectious human viruses in environmental samples without the need for primer design. This method can be expanded in future work to detect and quantify more types of culturable viruses and to address questions of virus fate and transmission in the environment.