Identification of Clinical Biomarkers for Pre-Analytical Quality Control of Blood Samples

Abstract

Background: Pre-analytical conditions are key factors in maintaining the high quality of biospecimens. They are necessary for accurate reproducibility of experiments in the field of biomarker discovery as well as achieving optimal specificity of laboratory tests for clinical diagnosis. In research at the National Biobank of Korea, we evaluated the impact of pre-analytical conditions on the stability of biobanked blood samples by measuring biochemical analytes commonly used in clinical laboratory tests. Methods: We measured 10 routine laboratory analytes in serum and plasma samples from healthy donors (n = 50) with a chemistry autoanalyzer (Hitachi 7600-110). The analyte measurements were made at different time courses based on delay of blood fractionation, freezing delay of fractionated serum and plasma samples, and at different cycles (0, 1, 3, 6, 9) of freeze-thawing. Statistically significant changes from the reference sample mean were determined using the repeated-measures ANOVA and the significant change limit (SCL). Results: The serum levels of GGT and LDH were changed significantly depending on both the time interval between blood collection and fractionation and the time interval between fractionation and freezing of serum and plasma samples. The glucose level was most sensitive only to the elapsed time between blood collection and centrifugation for blood fractionation. Based on these findings, a simple formula (glucose decrease by 1.387 mg/ dL per hour) was derived to estimate the length of time delay after blood collection. In addition, AST, BUN, GGT, and LDH showed sensitive responses to repeated freeze-thaw cycles of serum and plasma samples. Conclusion: These results suggest that GGT and LDH measurements can be used as quality control markers for certain pre-analytical conditions (eg, delayed processing or repeated freeze-thawing) of blood samples which are either directly used in the laboratory tests or stored for future research in the biobank.

Stability of 35 biochemical and immunological routine tests after 10 hours storage and transport of human whole blood at 21°C

“…Particular critical analytes are ALAT, K, LD, and P, which in most studies are reported as stable only for a few hours at room temperature and above (up to 12 h depending on analyte and study) [4,8–10,12,15,19]. …”

Inorganic Phosphorus and Potassium Are Putative Indicators of Delayed Separation of Whole Blood

“…These samples are obtained by centrifugation after anticoagulation (using an anticoagulant, such as EDTA or heparin) and coagulation of whole blood, respectively [4] . Currently, the delay between collection and whole-blood centrifugation can influence the concentrations of circulating blood components, including metabolites and cytokines, due to prolonged contact with cells 5 , 6 , 7 , 8 , 9 , 10 . Hemolysis, which results in the release of hemoglobin and other intracellular components from red blood cells, might also occur when whole-blood processing is delayed [11] .…”

“…In laboratory tests (such as clinical biochemistry and hematology) routinely performed for diagnosis or prediction of prognosis in a clinical laboratory, the main causes of laboratory errors are preanalytical variables involved with biospecimen collection, processing, and storage conditions 12 , 13 , 14 . Several studies demonstrated that delayed whole-blood separation affects the plasma and serum concentrations of biochemical analytes [e.g., alanine aminotransferase (ALT), aspartate aminotransferase (AST), inorganic phosphorus (IP), potassium (K + ), and lactate dehydrogenase (LDH)] 6 , 7 , 8 , 9 . In this study, we investigated whether clinical-biochemistry analytes can be used to assess the delayed whole-blood separation.…”

Digital pathology and image analysis augment biospecimen annotation and biobank quality assurance harmonization

“…PaxGene, Qiagen), may improve efficiencies at surgical/tissue collection sites. Considerable attention has been devoted to developing procedures for appropriate collection and optimum preservation methods including the time to preservation, sample sizes, fixation/ storage conditions and most recently, markers useful for assessing sample quality (6, 9-18). Such attention is reflected in the release of standard operating procedures (SOPs) by the National Cancer Institute Cancer Human Biobank (caHUB), which conducts tissue collection for The NIH Genotype-Tissue Expression (GTEx) Project (19).…”