TY - JOUR
T1 - Ensuring sample quality for blood biomarker studies in clinical trials
T2 - A multicenter international study for plasma and serum sample preparation
AU - Kong, Feng Ming
AU - Zhao, Lujun
AU - Wang, Luhua
AU - Chen, Yuhchyau
AU - Hu, Jie
AU - Fu, Xiaolong
AU - Bai, Chunxue
AU - Wang, Li
AU - Lawrence, Theodore S.
AU - Anscher, Mitchell S.
AU - Dicker, Adam
AU - Okunieff, Paul
N1 - Funding Information:
A variety of pre-analytic factors may affect biospecimen quality and thus biomarker results (28-30). This study was limited in that only sample selection, collection, and processing were studied, without consideration of variations in the physiological conditions of the participants, biospecimen delivery, freezing, storage, and thawing. Systematic studies in Biospecimen Science have been sponsored and conducted by programs such as the US National Cancer Institute’s Biospecimen Research Network [https://biospecimens.cancer.gov/ default.asp], and the Standardization and improvement of generic Pre-analytical tools and procedures for In-vitro DIAgnostics project (SPIDIA) [http://www.spidia. eu], a consortium that was funded by the European Union and coordinated by QIAGEN in Germany. We advise researchers particularly cooperative group for clinical trials to follow the best practice proposed by NCI (NCI Best practice for biospecimens https://biospecimens.cancer. gov/bestpractices/2016-NCIBestPractices.pdf), to develop evidence-based practices to guide biospecimen methodology to minimize confounding effect of pre-analytic factors, and mitigate pre-analytic effects when collecting and utilizing stored biospecimens.
Publisher Copyright:
© Translational lung cancer research.
PY - 2017/12/1
Y1 - 2017/12/1
N2 - Background: Sample quality is critical for biomarker detection in oncology, and platelet degradation and contamination in plasma have a remarkable impact on the ability to accurately quantify many blood-based biomarkers. Platelet factor 4 (PF4) can be used as an indicator to monitor sample quality. This multicenter study aimed to determine the impact of critical components of the blood sample handling process on platelet degradation/contamination and to establish an optimal method for collecting platelet-poor plasma samples. Methods: At each of six participating centers, blood samples were drawn from 12-13 healthy volunteers. Serum and plasma samples were prepared from whole blood samples using nine different methods that have been commonly used in ongoing multicenter trials. PF4 levels in the prepared samples were measured by enzyme-linked immunosorbent assay (ELISA). Paired t-tests were used for statistical analysis. Results: Blood samples were collected from 74 subjects enrolled in six centers. PF4 levels were significantly higher in serum samples than in plasma samples (P < 0.001), in plasma samples from blood that sat at room temperature for 5 minutes (P=0.021), in plasma samples prepared at an insufficient centrifugal force (P < 0.001), and in plasma samples prepared from blood that sat for longer than 4 hours on ice (P=0.001). For each method, the PF4 levels did not differ significantly among the centers or between Chinese and American subjects. The methods that resulted in normal levels of PF4 involved keeping blood samples on ice for 30 minutes to < 4 hours and centrifugation at 2,500-3,000 ×g for 30 min. Conclusions: This multicenter study evaluated multiple blood sample handling conditions for minimizing platelet degradation during plasma serum preparation and determined an optimal method for preparing platelet-poor plasma. The findings of this study can be applied in future blood biomarker studies.
AB - Background: Sample quality is critical for biomarker detection in oncology, and platelet degradation and contamination in plasma have a remarkable impact on the ability to accurately quantify many blood-based biomarkers. Platelet factor 4 (PF4) can be used as an indicator to monitor sample quality. This multicenter study aimed to determine the impact of critical components of the blood sample handling process on platelet degradation/contamination and to establish an optimal method for collecting platelet-poor plasma samples. Methods: At each of six participating centers, blood samples were drawn from 12-13 healthy volunteers. Serum and plasma samples were prepared from whole blood samples using nine different methods that have been commonly used in ongoing multicenter trials. PF4 levels in the prepared samples were measured by enzyme-linked immunosorbent assay (ELISA). Paired t-tests were used for statistical analysis. Results: Blood samples were collected from 74 subjects enrolled in six centers. PF4 levels were significantly higher in serum samples than in plasma samples (P < 0.001), in plasma samples from blood that sat at room temperature for 5 minutes (P=0.021), in plasma samples prepared at an insufficient centrifugal force (P < 0.001), and in plasma samples prepared from blood that sat for longer than 4 hours on ice (P=0.001). For each method, the PF4 levels did not differ significantly among the centers or between Chinese and American subjects. The methods that resulted in normal levels of PF4 involved keeping blood samples on ice for 30 minutes to < 4 hours and centrifugation at 2,500-3,000 ×g for 30 min. Conclusions: This multicenter study evaluated multiple blood sample handling conditions for minimizing platelet degradation during plasma serum preparation and determined an optimal method for preparing platelet-poor plasma. The findings of this study can be applied in future blood biomarker studies.
KW - Biomarker study
KW - Plasma
KW - Platelet factor 4 (PF4)
KW - Serum
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U2 - 10.21037/tlcr.2017.09.13
DO - 10.21037/tlcr.2017.09.13
M3 - Article
AN - SCOPUS:85031323584
SN - 2218-6751
VL - 6
SP - 625
EP - 634
JO - Translational Lung Cancer Research
JF - Translational Lung Cancer Research
IS - 6
ER -