Performance of Streck cfDNA Blood Collection Tubes for Liquid Biopsy Testing.
OBJECTIVE
Making liquid biopsy testing widely available requires a concept to ship whole blood at ambient temperatures while retaining the integrity of the cell-free DNA (cfDNA) population and stability of blood cells to prevent dilution of circulating tumor DNA (ctDNA) with wild-type genomic DNA. The cell- and DNA-stabilizing properties of Streck Cell-Free DNA BCT blood collection tubes (cfDNA BCTs) were evaluated to determine if they can be utilized in combination with highly sensitive mutation detection technologies.


METHODS
Venous blood from healthy donors or patients with advanced colorectal cancer (CRC) was collected in cfDNA BCTs and standard K2EDTA tubes. Tubes were stored at different temperatures for various times before plasma preparation and DNA extraction. The isolated cfDNA was analyzed for overall DNA yield of short and long DNA fragments using qPCR as well as for mutational changes using BEAMing and Plasma Safe-Sequencing (Safe-SeqS).


RESULTS
Collection of whole blood from healthy individuals in cfDNA BCTs and storage for up to 5 days at room temperature did not affect the DNA yield and mutation background levels (n = 60). Low-frequency mutant DNA spiked into normal blood samples as well as mutant circulating tumor DNA in blood samples from CRC patients collected in cfDNA BCTs were reliably detected after 3 days of storage at room temperature. However, blood samples stored at ≤ 10°C and at 40°C for an extended period of time showed elevated normal genomic DNA levels and an abnormally large cellular plasma interface as well as lower plasma volumes.


CONCLUSIONS
Whole blood shipped in cfDNA BCTs over several days can be used for downstream liquid biopsy testing using BEAMing and Safe-SeqS. Since the shipping temperature is a critical factor, special care has to be taken to maintain a defined room temperature range to obtain reliable mutation testing results.
Research and Development, Sysmex Inostics GmbH, Hamburg, Germany.
Inga
I
Medina Diaz
Research and Development, Sysmex Inostics GmbH, Hamburg, Germany.
Annette
A
Nocon
Research and Development, Sysmex Inostics GmbH, Hamburg, Germany.
Daniel H
DH
Mehnert
Research and Development, Sysmex Inostics GmbH, Hamburg, Germany.
Johannes
J
Fredebohm
Research and Development, Sysmex Inostics GmbH, Hamburg, Germany.
Frank
F
Diehl
Research and Development, Sysmex Inostics GmbH, Hamburg, Germany.
Frank
F
Holtrup
D001800:Blood Specimen Collection / Q000295:instrumentation / Q000379:methods*; D015179:Colorectal Neoplasms / Q000097:blood* / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D004273:DNA, Neoplasm / Q000097:blood* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D009154:Mutation; D013696:Temperature
D004273:DNA, Neoplasm