TY - JOUR
T1 - Comparison of cell stabilizing blood collection tubes for circulating plasma tumor DNA
AU - Toro, Patricia Valda
AU - Erlanger, Bracha
AU - Beaver, Julia
AU - Cochran, Rory L.
AU - VanDenBerg, Dustin A.
AU - Yakim, Elizabeth
AU - Cravero, Karen
AU - Chu, David
AU - Zabransky, Daniel J.
AU - Wong, Hong Yuen
AU - Croessmann, Sarah
AU - Parsons, Heather
AU - Hurley, Paula
AU - Lauring, Josh David
AU - Park, Ben Ho
N1 - Publisher Copyright:
© 2015 The Canadian Society of Clinical Chemists.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - Objectives: Circulating plasma DNA is being increasingly used for biomedical and clinical research as a substrate for genetic testing. However, cell lysis can occur hours after venipuncture when using standard tubes for blood collection, leading to an increase in contaminating cellular DNA that may hinder analysis of circulating plasma DNA. Cell stabilization agents can prevent cellular lysis for several days, reducing the need for immediate plasma preparation after venipuncture, thereby facilitating the ease of blood collection and sample preparation for clinical research. However, the majority of cell stabilizing reagents have not been formally tested for their ability to preserve circulating plasma tumor DNA. Design & methods: In this study, we compared the properties of two cell stabilizing reagents, the cell-free DNA BCT tube and the PAXgene tube, by collecting blood samples from metastatic breast cancer patients and measuring genome equivalents of plasma DNA by droplet digital PCR. We compared wild type PIK3CA genome equivalents and also assayed for two PIK3CA hotspot mutations, E545K and H1047R. Results: Our results demonstrate that blood stored for 7. days in BCT tubes did not show evidence of cell lysis, whereas PAXgene tubes showed an order of magnitude increase in genome equivalents, indicative of considerable cellular lysis. Conclusions: We conclude that BCT tubes can prevent lysis and cellular release of genomic DNA of blood samples from cancer patients when stored at room temperature, and could therefore be of benefit for blood specimen collections in clinical trials.
AB - Objectives: Circulating plasma DNA is being increasingly used for biomedical and clinical research as a substrate for genetic testing. However, cell lysis can occur hours after venipuncture when using standard tubes for blood collection, leading to an increase in contaminating cellular DNA that may hinder analysis of circulating plasma DNA. Cell stabilization agents can prevent cellular lysis for several days, reducing the need for immediate plasma preparation after venipuncture, thereby facilitating the ease of blood collection and sample preparation for clinical research. However, the majority of cell stabilizing reagents have not been formally tested for their ability to preserve circulating plasma tumor DNA. Design & methods: In this study, we compared the properties of two cell stabilizing reagents, the cell-free DNA BCT tube and the PAXgene tube, by collecting blood samples from metastatic breast cancer patients and measuring genome equivalents of plasma DNA by droplet digital PCR. We compared wild type PIK3CA genome equivalents and also assayed for two PIK3CA hotspot mutations, E545K and H1047R. Results: Our results demonstrate that blood stored for 7. days in BCT tubes did not show evidence of cell lysis, whereas PAXgene tubes showed an order of magnitude increase in genome equivalents, indicative of considerable cellular lysis. Conclusions: We conclude that BCT tubes can prevent lysis and cellular release of genomic DNA of blood samples from cancer patients when stored at room temperature, and could therefore be of benefit for blood specimen collections in clinical trials.
KW - Cell stabilizing tube
KW - Circulating tumor DNA
KW - Droplet digital PCR
KW - Plasma tumor DNA
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U2 - 10.1016/j.clinbiochem.2015.07.097
DO - 10.1016/j.clinbiochem.2015.07.097
M3 - Article
C2 - 26234639
AN - SCOPUS:84943588034
SN - 0009-9120
VL - 48
SP - 993
EP - 998
JO - Clinical Biochemistry
JF - Clinical Biochemistry
IS - 15
ER -