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THE DISCOVERY OF BIOMARKERS OF VIRAL INFECTIVITY BY MASS SPECTROMETRY
Citation:
Shoemaker, J A., J. M. Talley, D R. Dahling, AND G S. Fout. THE DISCOVERY OF BIOMARKERS OF VIRAL INFECTIVITY BY MASS SPECTROMETRY. Presented at Science Forum 2003, Washington, DC, May 5-7, 2003.
Impact/Purpose:
This particular task is comprised of 4 subtasks: 1.) Characterization of Potential Viral Biomarkers by Mass Spectrometry; 2.) Characterization of Parasites by Mass Spectrometric Techniques;
3.) Rapid Discrimination of Bacterial Indicators of Fecal Contamination and Bacterial Pathogens by Mass Spectrometric Techniques; and 4.) Investigation of Aeromonas Virulence Factors Using Mass Spectrometry.
The purpose of this research project is to use mass spectrometric techniques, such as electrospray ionization (ESI), capillary electrophoresis (CE) and matrix assisted laser desorption ionization (MALDI) mass spectrometry, to provide "protein mass fingerprinting" and protein sequencing information for viruses, bacteria and protozoa that cause waterborne disease. These protein mass fingerprinting libraries will be evaluated to determine whether mass spectrometric techniques can identify protein fingerprints related to the infectivity/viability of selected microorganisms and whether they can differentiate between infective / non-infective genus and strains of the selected microorganisms. The characteristic proteins identified by mass spectrometry as markers of infectivity/viability or strain differentiation can then be used to develop more sensitive microbiological drinking water methods.
Description:
Over the past three decades, the CDC and the U.S. EPA have collected and reported data relating to occurrences and causes of waterborne-disease outbreaks in the United States. Thirty nine outbreaks associated with drinking water were reported during 1999-2000. According to CDC's 1999-2000 water surveillance report, of the 20 outbreaks with known infectious causes, four were caused by viruses, which indicates that viral contamination of drinking water is a problem. A number of these viruses (e.g. coxsackie) are now listed on the U. S. EPA 1998 Contaminant Candidate List as needing additional research (i.e., analytical methods, occurrence, treatment and health effects). Coxsackievirus causes flu-like symptoms including diarrhea and fever and can cause complications such as diabetes mellitus and myocarditis. Thus, NERL is striving to develop methods for viruses listed on the CCL. This research project is a collaborative effort between NERL microbiologists and chemists to use analytical chemistry to differentiate between virus strains and to identify potential biomarkers of viral infectivity. Typical microbiological methods require months to determine the virus strain. Through the use of mass spectrometry, we have identified potential viral biomarkers which can be used to differentiate rapidly between the strains of coxsackievirus and can potentially indicate whether the virus is infectious. A unique peak was observed in an infectious coxsackievirus, but was not observed in a non-infectious coxsackievirus. This unique peak may be responsible for viral infectivity, thus, be a potential biomarker of infectivity. This type of information, gathered through the use of mass spectrometric techniques is cutting edge, ground breaking basic research, and can have a significant impact on the field of virology and play a role in the future development of drinking water methods to detect viruses and their infectivity.