Eric Delwart, Ph.D.
Senior Investigator, BSRI
Professor, Department of Laboratory Medicine
University of California, San Francisco
Viral Metagenomic Analysis of Plasma from CFS patients
Eric Delwart, Ph.D., University of California San Francisco and Blood Systems Research Institute is cataloging all the viruses, or virome, from 200 ME/CFS blood samples from the SolveCFS BioBank. He will look for novel viruses as well as compare those found to what is known in healthy blood samples. He will do this by using filtering the plasma to remove bacteria and human cells. The filtrate is treated with a cocktail of nuclease to digest non-viral capsid protected (mostly human and bacterial) nucleic acids. The RNA and DNA protected within viral capsids are then extracted and amplified in a non-biased way using random RT-PCR. The random amplification products are then deep sequenced using 454 pyrosequencing or Illumina sequencing. The resulting genetic information is then analyzed using a super-computer for similarities to known human and animal viruses in order to detect novel viral-like sequences. Viral-like sequences are used as a genetic foothold to sequence the entire viral genome. Knowing the viral genome allows sensitive PCR to test other samples for the presence of that viral sequence. Viral prevalence is then compared in symptomatic cases versus epidemiologically matched healthy controls to determine if the presence of the virus is associated with CFS.