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10.1128/JCM.00649-21

http://scihub22266oqcxt.onion/10.1128/JCM.00649-21
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34550806!8601232!34550806
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suck abstract from ncbi


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pmid34550806      J+Clin+Microbiol 2021 ; 59 (12): e0064921
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  • Whole-Genome Sequencing of SARS-CoV-2: Assessment of the Ion Torrent AmpliSeq Panel and Comparison with the Illumina MiSeq ARTIC Protocol #MMPMID34550806
  • Plitnick J; Griesemer S; Lasek-Nesselquist E; Singh N; Lamson DM; St George K
  • J Clin Microbiol 2021[Nov]; 59 (12): e0064921 PMID34550806show ga
  • Fast and effective methods are needed for sequencing of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome to track genetic mutations and to identify new and emerging variants during the ongoing pandemic. The objectives were to assess the performance of the SARS-CoV-2 AmpliSeq research panel and S5 plug-in analysis tools for whole-genome sequencing analysis of SARS-CoV-2 and to compare the results with those obtained with the MiSeq-based ARTIC analysis pipeline, using metrics such as depth, coverage, and concordance of single-nucleotide variant (SNV) calls. A total of 191 clinical specimens and a single cultured isolate were extracted and sequenced with AmpliSeq technology and analysis tools. Of the 191 clinical specimens, 83 (with threshold cycle [C(T)] values of 15.58 to 32.54) were also sequenced using an Illumina MiSeq-based method with the ARTIC analysis pipeline, for direct comparison. A total of 176 of the 191 clinical specimens sequenced on the S5XL system and prepared using the SARS-CoV-2 research panel had nearly complete coverage (>98%) of the viral genome, with an average depth of 5,031x. Similar coverage levels (>98%) were observed for 81/83 primary specimens that were sequenced with both methods tested. The sample with the lowest viral load (C(T) value of 32.54) achieved 89% coverage using the MiSeq method and failed to sequence with the AmpliSeq method. Consensus sequences produced by each method were identical for 81/82 samples in areas of equal coverage, with a single difference present in one sample. The AmpliSeq approach is as effective as the Illumina-based method using ARTIC v3 amplification for sequencing SARS-CoV-2 directly from patient specimens across a range of viral loads (C(T) values of 15.56 to 32.54 [median, 22.18]). The AmpliSeq workflow is very easily automated with the Ion Chef and S5 instruments and requires less training and experience with next-generation sequencing sample preparation than the Illumina workflow.
  • |*COVID-19[MESH]
  • |*SARS-CoV-2[MESH]
  • |Genome, Viral/genetics[MESH]
  • |High-Throughput Nucleotide Sequencing[MESH]
  • |Humans[MESH]
  • |Pandemics[MESH]


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  • suck abstract from ncbi

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