Selected article for: "additional evidence and phylogenetic analysis"
Author: Chenyu Li; David N. Debruyne; Julia Spencer; Vidushi Kapoor; Lily Y. Liu; Bo Zhou; Lucie Lee; Rounak Feigelman; Grayson Burdon; Jeffrey Liu; Alejandra Oliva; Adam Borcherding; Hongdong Tan; Alexander E. Urban; Guoying Liu; Zhitong Liu
Title: High sensitivity detection of coronavirus SARS-CoV-2 using multiplex PCR and a multiplex-PCR-based metagenomic method
  • Document date: 2020_3_14
    • ID: 0hxan9rw_29
    Snippet: Here, we report the development of a multiplex PCR assay for high-sensitivity identification of SARS-CoV-2 infection. With 172 pairs of primers, this method enables the detection of low copy numbers, potentially degraded fragments of the viral genome, or even a mutated variant, with high confidence. The 172 pairs of primers cover about 56% of the genome of SARS-CoV-2. When detecting limited copies of virus, fewer targets are successfully amplifie.....
    Document: Here, we report the development of a multiplex PCR assay for high-sensitivity identification of SARS-CoV-2 infection. With 172 pairs of primers, this method enables the detection of low copy numbers, potentially degraded fragments of the viral genome, or even a mutated variant, with high confidence. The 172 pairs of primers cover about 56% of the genome of SARS-CoV-2. When detecting limited copies of virus, fewer targets are successfully amplified. In the case of one copy of virus, we estimate that 20% of the viral genome, or 6kb of sequences, could be amplified. These are sufficient to produce a conspicuous peak in electrophoresis for an initial indication of positives. The use of NGS sequencing can provide nucleic acid-level information to further confirm the identification, and provide additional evidence for phylogenetic analysis. In addition, our method is robust, accurate and easily performed from different levels of expertise in various laboratory settings.

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