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10.1016/j.jmoldx.2021.05.016

http://scihub22266oqcxt.onion/10.1016/j.jmoldx.2021.05.016
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suck abstract from ncbi


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pmid34153515      J+Mol+Diagn 2021 ; 23 (9): 1065-1077
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  • Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Genome Sequencing from Post-Mortem Formalin-Fixed, Paraffin-Embedded Lung Tissues #MMPMID34153515
  • Van Campenhout C; De Mendonca R; Alexiou B; De Clercq S; Racu ML; Royer-Chardon C; Rusu S; Van Eycken M; Artesi M; Durkin K; Mardulyn P; Bours V; Decaestecker C; Remmelink M; Salmon I; D'Haene N
  • J Mol Diagn 2021[Sep]; 23 (9): 1065-1077 PMID34153515show ga
  • Implementation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing in the daily practice of pathology laboratories requires procedure adaptation to formalin-fixed, paraffin-embedded (FFPE) samples. So far, one study reported the feasibility of SARS-CoV-2 genome sequencing on FFPE tissues with only one contributory case of two. This study optimized SARS-CoV-2 genome sequencing using the Ion AmpliSeq SARS-CoV-2 Panel on 22 FFPE lung tissues from 16 deceased coronavirus disease 2019 (COVID-19) patients. SARS-CoV-2 was detected in all FFPE blocks using a real-time RT-qPCR targeting the E gene with crossing point (Cp) values ranging from 16.02 to 34.16. Sequencing was considered as contributory (i.e. with a uniformity >55%) for 17 FFPE blocks. Adapting the number of target amplification PCR cycles according to the RT-qPCR Cp values allowed optimization of the sequencing quality for the contributory blocks (i.e. 20 PCR cycles for blocks with a Cp value <28 and 25 PCR cycles for blocks with a Cp value between 28 and 30). Most blocks with a Cp value >30 were non-contributory. Comparison of matched frozen and FFPE tissues revealed discordance for only three FFPE blocks, all with a Cp value >28. Variant identification and clade classification was possible for 13 patients. This study validates SARS-CoV-2 genome sequencing on FFPE blocks and opens the possibility to explore correlation between virus genotype and histopathologic lesions.
  • |Autopsy[MESH]
  • |COVID-19/pathology/*virology[MESH]
  • |Formaldehyde[MESH]
  • |Genome, Viral/*genetics[MESH]
  • |High-Throughput Nucleotide Sequencing/methods[MESH]
  • |Humans[MESH]
  • |Lung/pathology/*virology[MESH]
  • |Paraffin Embedding[MESH]
  • |Real-Time Polymerase Chain Reaction/*methods[MESH]
  • |SARS-CoV-2/*genetics/isolation & purification[MESH]


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