Frequently Asked Questions

  • How does DU maintain sample integrity and chain of custody?

    Each patient, upon arrival, is asked to scan their DU ID and show picture identification. Two forms of identification are placed in each sample in addition to the vial bar code (Name and Date of Birth). This information is both logged in the myhealth medical record system and on paper backup logs. The process is entirely contactless to promote the safest collection environment. Privacy screens are provided for those who wish to use them. The patient places their own vial in the transport cooler. Each cooler is numbered and tracked when it leaves the collection site, with information about who transported it, when it arrived at the lab, and who received it. Vials are then scanned into the lab software connecting a sample ID with a plate number and well position. These files are then sent to the HCC for confirmation that all samples are registered in the system and any issues are corrected via the paper logs.  

  • What are the actual techniques deployed to analyze samples?

    Extraction: When the DU SPIT receives a saliva sample, the first step is to isolate genetic material in order to look for SAR-CoV-2 virus. The University of Denver uses an RNA Advance kit from Beckman Coulter and robotic handling to purify the saliva by extracting RNA and RNA fragments using magnetic beads into a fresh fluid. The RNA material adheres to the bead and those beads are washed multiple times to remove saliva and other materials found in the saliva. The extracted RNA is then released or eluded from the beads in one step into a fresh buffer.     

    Amplification Technique: The concentrated RNA solution is then mixed with the COVID-19 Taqpath kit. This kit uses molecular probes to adhere to three specific COVID-19 genes: Orf, N, and Spike. These molecular probes fluoresce when excited and concentration is measured easily as a function of light intensity in real-time. A classical reverse transcription methodology is employed to replicate the RNA and RNA fragments. Each replication cycle is referred to as “cycle” and the quant studio machines record the increase in fluoresce of each of the three genes with each cycle. At the end of 40+ cycles, if all three genes are present, the lab quantifies the sample as positive and calculates the viral load or copies of viral RNA per ml based on the fluorescence.    

  • Does the University perform quality assurance on the test results?

    The SPIT Lab uses in-plate and in-well controls to determine if the sample results reported meet quality controls. Our in-well control is associated with a virus, thus providing a better comparison if the in-well replication process worked. We use all three genetic markers instead of one marker like many other labs. We have clear thresholds based on optical limits of detection and analytical thresholds that must be exceeded before a positive result is reported.   

  • Can the SPIT Lab assess any of the variants?

    If a sample is positive for the genes associated with SARS-CoV-2, the extracted sample is processed using a melt curve analysis. Most labs sequence the concentrated material to understand the variant. This is time-consuming and expensive. DU has leveraged the fact that variations in genetic sequence in an RNA material will show differences in a melt curve. We have chosen to use this methodology to determine and report the variant to the state.  

  • Does the SPIT Lab maintain my genetic information from the saliva test?

    No, the primary saliva sample is destroyed approximately 24 hours after collection. This is articulated in our lab protocols but is actually a requirement for the lab to function. There is simply not enough room to store 1,200 samples per day, seven days a week, for very long. The extracted RNA samples are also destroyed but all positive samples are consumed in the melt curve analysis.   

  • What types of lab certifications or approvals has the SPIT Lab received?

    The lab is CLIA and CAP certified. These certifications define how the lab operates from sample collection to destruction. DU has also decided to have oversight of the protocols reviewed, monitored and approved by the Institutional Human Subjects review board. This is a collection of experts charged with ensuring the protocols that the lab utilizes place the needs of the human subjects first and minimizes all risks. This includes the storage and handling of private health information and materials. Additionally, DU’s SPIT lab has an EUA application in with the FDA. Our application is under evaluation based on the questions we have received and answered but we do not know when approval might be granted.   

  • How do the SPIT Lab and the COVID team protect private health information?

    The University of Denver is bound by FERPA regulations, but as an enhanced requirement, DU also applies HIPPA to all testing and medical data stored in the myhealth system in order to protect the privacy of its students, staff, and employees. All medical information from salivary testing is automatically transferred to HCC medical records once the samples have been resulted and analyzed.