To identify and quantify the virus we need to run several PCR steps with different reagents on multiple instruments. This is because the genetic material of the virus is RNA instead of DNA. DNA is relatively easy. For example, with our mold analyses (ERMI, SIM, Big 2, SIAM etc.), we place the mold capture medium in a small tube with tiny glass beads and beat them vigorously in an apparatus that’s a little like a paint mixer, which cracks open the mold spores. Then we just drain out the DNA after washing away contaminants like proteins and carbohydrates and analyze it. This will not work for RNA. RNA is a wimpy nucleic acid compared to DNA. I often joke in the lab “don’t breathe on the RNA or it will fall apart.” Hence, it’s impossible to use disruptive bead beating for RNA extraction, otherwise the end product is completely sheared into tiny bits that are impossible to decode with PCR. Instead we have to lyse the virus with gentle chemicals and carefully elute the RNA for downstream purification and PCR. It’s like that old game we played as kids with an egg on a spoon where you had to walk to the finish line without dropping.
Once we have the RNA purified the fun starts. We cannot decode it directly, but science has found a work around. We take the RNA and run a conversion PCR method called RT-PCR or Reverse Transcriptase PCR. Note, for those of you not familiar with PCR, it stands for Polymerase Chain Reaction. It allows scientists to detect nucleic acids, RNA and DNA. We need those nucleic acids so we can genetically decode them to identify specific genes, or in our case unique strains and species of microbes. The problem is that if we have one virus or one mold spore the amount of nucleic acids in that cell is not concentrated enough for our current technology to detect. The work around is to concentrate it. That is why I often call PCR a molecular photocopy machine. It takes a single copy of DNA and turns it into millions of copies in a relatively short period of time. It’s easy for lab instrumentation to detect millions of copies, and decode them to identify a microbial target.
Back to RNA. In order to decode the RNA we use RT-PCR which takes the RNA genetic code and converts it into millions of copies of its complementary DNA strands. Remember we cannot read RNA directly as it would fall apart during the copying process. By converting it to DNA we end up with a stable nuclei acid that is easy to decode because it is a mirror image of the original RNA sequence. Once we convert that target into DNA, we can concentrate and measure it using quantitative PCR (qPCR) in several simple calculations.
To wrap up this discussion, Assured Bio has significantly increased its equipment and capacity to handle COVID-19 testing. We are awaiting one final approval by our accrediting body, the American Industrial Hygiene Associations, to become the first AIHA accredited COVID-19 environmental lab in North America for surfaces and air. In terms of testing, we have been running samples since March for local municipalities and for NYC essential businesses in the financial district. We recently started testing Long Term Care Facilities to provide management with the data they need to protect our most vulnerable group of Americans. Moreover, we are providing COVID-19 surface testing and air testing to healthcare facilities in the northeast U.S. as they are converting COVID-19 patient rooms back into standard patient rooms. We deliver same day turnaround during week days and upon request for weekends. Assured Bio is committed to supporting our clients during these trying times.
As the country prepares for mass employee re-entry in some of the hardest hit locations, we will provide weekly testing and valuable cleaning methodology to employers so they may keep employees safe. We are deploying our WhisperCareTM continuous air monitoring systems for COVID-19 air testing. We are currently using WhisperCareTM systems in firehouses to quickly identify the presence of COVID-19, and prevent widespread infection of crucial first responders. Commercial clients are currently assessing their facilities and working with us to determine the best locations to place the monitors, and the frequency of reporting while verifying cleaning efficacy. In short, we have many unknowns ahead, but I feel confident that by using common sense, good science and savvy technology, workplace re-entry is going to be a success even in the worst hit locations.
Edward Sobek, PhD MBA
Founder/Principal Assured Bio labs, LLC