Selected article for: "dilution series and SARS spike"

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_42
    Snippet: qSanger is a quantitative approach to measuring viral abundance in a sample. ( A ) Data analysis and interpretation in a qSanger COVID-19 assay. Representative Sanger sequencing chromatograms are shown for amplified products of spike-in DNA only, SarS-CoV-2 RNA only, or a mixture of the two. Since the spike-in is an internal control for amplification and sequencing, observing the spike-in signal only indicates that SARS-CoV-2 RNA was absent, and .....
    Document: qSanger is a quantitative approach to measuring viral abundance in a sample. ( A ) Data analysis and interpretation in a qSanger COVID-19 assay. Representative Sanger sequencing chromatograms are shown for amplified products of spike-in DNA only, SarS-CoV-2 RNA only, or a mixture of the two. Since the spike-in is an internal control for amplification and sequencing, observing the spike-in signal only indicates that SARS-CoV-2 RNA was absent, and therefore the specimen is negative for COVID-19. In contrast, observing SARS-CoV-2 signal only indicates that SARS-COV-2 RNA was so abundant in the specimen that it is above the quantifiable range. In a mixed chromatogram of spike-in and SARS-CoV-2, the abundance of SARS-CoV-2 RNA is determined by the relative contributions of SARS-CoV-2 and spike-in signal intensities. ( B -D ) Synthetic viral genomic RNA was added to RT-PCR reactions at 10, 100, 1000, and 5000 GCE. The same dilutions were subjected to qSanger testing and RT-qPCR. ( B ) RT-qPCR exhibits a linear estimate across the dilution series, consistent with previous results. ( C ) Across the same dilution series, the ratio (R) of genomic sequence to spike-in sequence scales with RNA added. ( D ) When the qPCR estimate of abundance is compared to qSanger estimates of abundance, they exhibit a strong linear relationship, indicating that qSanger performs as well as qPCR in estimates of viral RNA abundance. author/funder. All rights reserved. No reuse allowed without permission.

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