Selected article for: "copy reaction and genome copy"
Author: Devon Chandler-Brown; Anna M. Bueno; Oguzhan Atay; David S. Tsao
Title: A Highly Scalable and Rapidly Deployable RNA Extraction-Free COVID-19 Assay by Quantitative Sanger Sequencing
  • Document date: 2020_4_10
    • ID: eui41zyg_18
    Snippet: Here, we describe a novel quantitative Sanger (qSanger) assay that can detect COVID-19 without RNA extraction. We show that qSanger performs as well as qPCR in estimates of viral RNA abundance and consistently detects as low as 10-20 viral genome copy equivalents, even when VTM is added directly into the reaction mix without RNA extraction. Because qSanger is an end-point PCR reaction with an internal spike-in control, it is more robust to inhibi.....
    Document: Here, we describe a novel quantitative Sanger (qSanger) assay that can detect COVID-19 without RNA extraction. We show that qSanger performs as well as qPCR in estimates of viral RNA abundance and consistently detects as low as 10-20 viral genome copy equivalents, even when VTM is added directly into the reaction mix without RNA extraction. Because qSanger is an end-point PCR reaction with an internal spike-in control, it is more robust to inhibitors that can exist in VTM, and failures in amplification result in undetermined results that require a repeat reaction, as opposed to false negatives that would be obtained by qPCR. It also has higher specificity than qPCR, as the examination of the sequencing information can be used to distinguish similar viruses and rule out false-positives due to non-specific amplifications.

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