Plasma ctDNA was evaluated by the Huajianwei bespoke MRD assay based on Signatera as previously described13 (link),31 (link). Briefly, 16 top-ranked SNVs were selected based on the tumor tissue whole exome sequencing (WES) data generated from a pan-cancer WES panel (Quanxi) on the MGISEQ-2000 platform (MGI Tech), and 16-plex specific primer pairs were used to amplify the universal cfDNA libraries. The products were then sequenced on the HiSeq2500 system (Illumina Inc). Detailed methods are described in the Supplemental Digital Content 2, http://links.lww.com/JS9/C41 . To minimize the false positive rate, the plasma sample with at least two variants positive (out of 16 variants in total per person) was defined as ctDNA or MRD positive, and ctDNA was quantified in mean tumor molecule per milliliter (MTM/ml) plasma (Supplementary eTable 3, Supplemental Digital Content 3, http://links.lww.com/JS9/C42 ). The raw data that support the findings of this study have been deposited into CNGB Sequence Archive (CNSA)32 of China National GeneBank DataBase (CNGBdb)33 (link) with accession number CNP0005016 (https://db.cngb.org/ ).
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Plasma ctDNA Evaluation by Signatera Assay
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Corresponding Organization :
Other organizations : Shanghai Medical College of Fudan University, BGI Group (China), Tianjin Medical University, Cancer Institute (WIA), Fudan University Shanghai Cancer Center
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Variable analysis
independent variables
- Tumor tissue whole exome sequencing (WES) data generated from a pan-cancer WES panel (Quanxi) on the MGISEQ-2000 platform
- Presence or absence of at least two variants positive (out of 16 variants in total per person) in the plasma sample, which was defined as ctDNA or MRD positive
- Quantification of ctDNA in mean tumor molecule per milliliter (MTM/ml) plasma
- The use of 16-plex specific primer pairs to amplify the universal cfDNA libraries
- The sequencing of the products on the HiSeq2500 system (Illumina Inc)
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