Yunlong Zhao1, Yingjie Li1, Ping Chen1, Shaojun Li1, Jian Luo2, Hui Xia1. 1. Department of Cardiothoracic Surgery, First Affiliated Hospital of PLA General Hospital, Beijing, China. 2. Burning Rock Biotech, Guangzhou, China.
Abstract
BACKGROUND: The ratio of target cfDNA in total plasma is low. The abundant gDNA background resulting from blood cell lysis caused by improper operation has become a major obstacle to accurately measure cfDNA. In this study, we investigated the storage capacity of three blood cell collection tubes (BCTs) in the prevention of genomic DNA contamination caused by white blood cell rupture and evaluated their performance when they were utilized combining with highly sensitive mutation detection technology. METHODS: Blood samples were drawn from six healthy blood donors and stored in three types of BCTs (BD K2 -EDTA tube, Roche tube, and Streck tube). Plasma samples were isolated at specific time points (day 0, day 3, day 7, and day 14) for content analysis. RESULTS: Roche BCT was more capable for preventing cfDNA contamination caused by white blood cell disruption within 14 days, comparing with BD K2 -EDTA tube and Streck tube. Severe white blood cell lysis and gDNA contamination were found in the BD tube. The impacts of Roche and Streck tubes on the quantity and complexity of next-generation sequencing (NGS) libraries did not differ significantly within 3 days, satisfying most of our daily demands. In addition, the rupture of WBC was not synchronized with hemolysis in BCTs. CONCLUSION: This study showed that capacities of blood collection tubes differed considerably in preservation of blood samples. Therefore, suitable blood collection devices should be selected to minimize gDNA contamination and to standardize blood samples processing for achieving more accurate and reliable clinical analysis of cfDNA.
BACKGROUND: The ratio of target cfDNA in total plasma is low. The abundant gDNA background resulting from blood cell lysis caused by improper operation has become a major obstacle to accurately measure cfDNA. In this study, we investigated the storage capacity of three blood cell collection tubes (BCTs) in the prevention of genomic DNA contamination caused by white blood cell rupture and evaluated their performance when they were utilized combining with highly sensitive mutation detection technology. METHODS: Blood samples were drawn from six healthy blood donors and stored in three types of BCTs (BD K2 -EDTA tube, Roche tube, and Streck tube). Plasma samples were isolated at specific time points (day 0, day 3, day 7, and day 14) for content analysis. RESULTS: Roche BCT was more capable for preventing cfDNA contamination caused by white blood cell disruption within 14 days, comparing with BD K2 -EDTA tube and Streck tube. Severe white blood cell lysis and gDNA contamination were found in the BD tube. The impacts of Roche and Streck tubes on the quantity and complexity of next-generation sequencing (NGS) libraries did not differ significantly within 3 days, satisfying most of our daily demands. In addition, the rupture of WBC was not synchronized with hemolysis in BCTs. CONCLUSION: This study showed that capacities of blood collection tubes differed considerably in preservation of blood samples. Therefore, suitable blood collection devices should be selected to minimize gDNA contamination and to standardize blood samples processing for achieving more accurate and reliable clinical analysis of cfDNA.
Authors: Sonya Parpart-Li; Bjarne Bartlett; Maria Popoli; Vilmos Adleff; Laura Tucker; Rebecca Steinberg; Andrew Georgiadis; Jill Phallen; Julie Brahmer; Nilo Azad; Ilene Browner; Daniel Laheru; Victor E Velculescu; Mark Sausen; Luis A Diaz Journal: Clin Cancer Res Date: 2016-11-08 Impact factor: 12.531
Authors: G D Sorenson; D M Pribish; F H Valone; V A Memoli; D J Bzik; S L Yao Journal: Cancer Epidemiol Biomarkers Prev Date: 1994 Jan-Feb Impact factor: 4.254
Authors: Inga Medina Diaz; Annette Nocon; Daniel H Mehnert; Johannes Fredebohm; Frank Diehl; Frank Holtrup Journal: PLoS One Date: 2016-11-10 Impact factor: 3.240
Authors: Laure Sorber; Karen Zwaenepoel; Julie Jacobs; Koen De Winne; Kaat Van Casteren; Elien Augustus; Filip Lardon; Hans Prenen; Marc Peeters; Jan Van Meerbeeck; Geert Roeyen; Christian Rolfo; Patrick Pauwels Journal: Mol Diagn Ther Date: 2020-02 Impact factor: 4.074
Authors: Sarah R Greytak; Kelly B Engel; Sonya Parpart-Li; Muhammed Murtaza; Abel J Bronkhorst; Mark D Pertile; Helen M Moore Journal: Clin Cancer Res Date: 2020-03-02 Impact factor: 12.531
Authors: Jacob E Till; Taylor A Black; Caren Gentile; Aseel Abdalla; Zhuoyang Wang; Hareena K Sangha; Jacquelyn J Roth; Robyn Sussman; Stephanie S Yee; Mark H O'Hara; Jeffrey C Thompson; Charu Aggarwal; Wei-Ting Hwang; Kojo S J Elenitoba-Johnson; Erica L Carpenter Journal: J Mol Diagn Date: 2021-08-25 Impact factor: 5.341