Evaluating transport conditions of conventional, widely used EDTA blood tubes for gene expression analysis in comparison to expensive, specialized PAXgene tubes in preparedness for radiological and nuclear events

Abstract

Purpose

Gene expression (GE) analysis of a radio-sensitive gene set (FDXR, DDB2, WNT3, POU2AF1) has been introduced in the last decade as an early and high-throughput prediction tool of later developing acute hematologic radiation syndrome (H-ARS) severity. The use of special tubes for RNA extraction from peripheral whole blood (PAXgene) represent an established standard in GE studies, although uncommonly used in clinics and not immediately available in the quantities needed in radiological/nuclear (R/N) incidents. On the other hand, EDTA blood tubes are widely utilized in clinical practice.

Material and Methods

Using blood samples from eleven healthy donors, we investigated GE changes associated with delayed processing of EDTA tubes up to 4 h at room temperature (RT) after venipuncture (simulating delays caused by daily clinical routine), followed by a subsequent transport time of 24 h at RT, 4 °C, and −20 °C. Differential gene expression (DGE) of the target genes was further examined after X-irradiation with 0 Gy and 4 Gy under optimal transport conditions.

Results

No significant changes in DGE were observed when storing EDTA whole blood samples up to 4 h at RT and subsequently kept at 4 °C for 24 h which is in line with expected DGE. However, other storage conditions, such as −20 °C or RT, decreased RNA quality and/or (significantly) caused changes in DGE exceeding the known methodological variance of the qRT-PCR.

Conclusion

Our data indicate that the use of EDTA whole blood tubes for GE-based H-ARS severity prediction is comparable to the quality of PAXgene tubes, when processed ≤ 4 h after venipuncture and the sample is transported within 24 hours at 4 °C.

Keywords:

• Peripheral EDTA whole blood
• qRT-PCR
• biomarker
• H-ARS severity prediction
• transport conditions

Disclosure statement

The authors report there are no competing interests to declare.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary materials.

Additional information

Funding

The German Ministry of Defense funded the work.

Notes on contributors

Simone Schüle

Simone Schüle, MD, is a Post-Doctoral Researcher of Radiobiology and a resident in Radiology at the Bundeswehr Institute of Radiobiology, Munich, Germany.

Patrick Ostheim

Patrick Ostheim, MD, is a Post-Doctoral Researcher of Radiobiology and a resident in Radiology at the Bundeswehr Institute of Radiobiology, Munich, Germany.

Razan Muhtadi

Razan Muhtadi, M.Sc., has a Master’s Degree in Radiation Biology and is a doctoral student at the Bundeswehr Institute of Radiobiology, Munich, Germany.

Samantha Stewart

Samantha Stewart, M.Sc., has a Master’s Degree in Radiation Biology and is a doctoral student at the Bundeswehr Institute of Radiobiology, Munich, Germany.

Gwendolyn Kaletka

Gwendolyn Kaletka, M.Sc., has a Master’s Degree in Radiation Biology and was a doctoral student at the Bundeswehr Institute of Radiobiology, Munich, Germany.

Cornelius Hermann

Cornelius Hermann, is a pharmacist and researcher in radiobiology at the Bundeswehr Insitute of Radiobiology, Munich, Germany.

Matthias Port

Matthias Port, MD, is a Professor of Radiobiology and Internal Medicine and Head of the Bundeswehr Institute of Radiobiology, Munich, Germany.

Michael Abend

Michael Abend, MD, is a Professor of Radiobiology and Deputy Head of the Bundeswehr Institute of Radiobiology, Munich, Germany.