Feng-Ming Spring Kong1, Lujun Zhao2, Luhua Wang3, Yuhchyau Chen4, Jie Hu5, Xiaolong Fu6, Chunxue Bai5, Li Wang7, Theodore S Lawrence8, Mitchell S Anscher7, Adam Dicker9, Paul Okunieff10. 1. Department of Radiation Oncology, Indiana University School of Medicine, Indianapolis, IN, USA. 2. Department of Radiation Oncology, Tianjin Medical University Cancer Hospital, Tianjin 300060, China. 3. Department of Radiation Oncology, Cancer Institute and Hospital, Chinese Academy of Science, Peking Union Medical College, Beijing 100021, China. 4. Deapartment of Radiation Oncology, University of Rochester, Rochester, NY, USA. 5. Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China. 6. Department of Radiation Oncology, Chest Hospital, Jiaotong University, Shanghai 200030, China. 7. Department of Radiation Oncology, UT MD Anderson Cancer Center, Houston, TX, USA. 8. Department of Radiation Oncology, University of Michigan, Ann Arbor, Michigan, USA. 9. Department of Radiation Oncology, Thomas Jefferson Hospital, Philadelphia, PA, USA. 10. Department of Radiation Oncology, University of Florida, Gainesville, FL, USA.
Abstract
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.
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.
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