gdna contamination
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2019 ◽  
Vol 21 (Supplement_2) ◽  
pp. ii88-ii88
Author(s):  
Agda Karina Eterovic ◽  
Nader Ezzeddine ◽  
Ping Song ◽  
Lin-ya Tang ◽  
Mohammad Mohammad ◽  
...  

Cancers ◽  
2018 ◽  
Vol 10 (9) ◽  
pp. 290 ◽  
Author(s):  
Laure Sorber ◽  
Karen Zwaenepoel ◽  
Koen De Winne ◽  
Kaat Van Casteren ◽  
Elien Augustus ◽  
...  

A multicenter study was performed to determine an optimal workflow for liquid biopsy in a clinical setting. In total, 549 plasma samples from 234 non-small cell lung cancer (NSCLC) patients were collected. Epidermal Growth Factor Receptor (EGFR) circulating cell-free tumor DNA (ctDNA) mutational analysis was performed using digital droplet PCR (ddPCR). The influence of (pre-) analytical variables on ctDNA analysis was investigated. Sensitivity of ctDNA analysis was influenced by an interplay between increased plasma volume (p < 0.001) and short transit time (p = 0.018). Multistep, high-speed centrifugation both increased plasma generation (p < 0.001) and reduced genomic DNA (gDNA) contamination. Longer transit time increased the risk of hemolysis (p < 0.001) and low temperatures were shown to have a negative effect. Metastatic sites were found to be strongly associated with ctDNA detection (p < 0.001), as well as allele frequency (p = 0.034). Activating mutations were detected in a higher concentration and allele frequency compared to the T790M mutation (p = 0.003, and p = 0.002, respectively). Optimization of (pre-) analytical variables is key to successful ctDNA analysis. Sufficient plasma volumes without hemolysis or gDNA contamination can be achieved by using multistep, high-speed centrifugation, coupled with short transit time and temperature regulation. Metastatic site location influenced ctDNA detection. Finally, ctDNA levels might have further value in detecting resistance mechanisms.


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