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

Author(s): Kang HJ, Jeon SY, Park JS, Yun JY, Kil HN, Hong WK, Lee MH, Kim JW, Jeon JP, Han BG

Publication: Biopreserv Biobank, 2013, Vol. 11, Page 94-100

PubMed ID: 23634248 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of delayed blood separation, delayed freezing and freeze-thaw cycling on levels of 10 clinical chemistry analytes in serum from healthy volunteers. For freeze-thaw cycling experiments, serum was frozen at -196 degrees C, thawed at room temperature, and returned to -196 degrees C. Clinical relevance was determined using the significant change limit (SCL).

   Summary of Findings:

   When whole blood was stored in SST tubes at room temperature, serum levels of AST, GGT, LDH, and creatinine levels increased significantly after 1 h (p<0.05, all), ALT and BUN increased after 4 h (p<0.05, both), glucose decreased significantly after 1 h (p<0.05), and total cholesterol (TC) was significantly elevated after 4 h but lower after 24 h. However, only the changes in GGT, LDH and glucose after at least a 2 h delay before blood separation were considered clinically relevant. All of the analytes affected by delayed centrifugation were also affected when serum was stored at room temperature for 2 h before freezing instead of frozen after 0.5 h, but storage of serum led to higher, not lower glucose and increases in TC and triglycerides (TG). However, the delayed freezing of serum only caused clinically relevant changes in GGT, LDH, and BUN. Compared to fresh serum, serum that was subjected to 1 freeze-thaw cycle had higher levels of ALT, AST, GGT, LDH, BUN, creatinine, and glucose (all p<0.05), and the changes in AST, GGT, LDH, and BUN were considered clinically relevant. AST, ALT, LDH, BUN and glucose levels tended to increase further with additional freeze-thaw cycles, and TC and TG decreased with 3 or more and 3 freeze-thaw cycles, respectively. Importantly, CRP levels in serum were not affected by storage of whole blood or serum at room temperature or subjecting serum to up to 9 freeze-thaw cycles. Glucose and creatinine levels were correlated with length of centrifugation delay (R=0.846 and R=0.407, respectively), and LDH levels were weakly correlated with the number of freeze-thaw cycles that serum was subjected to (R=0.251).

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Serum

   Preservative Types

   • None (Fresh)
   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Carbohydrate	Clinical chemistry/auto analyzer
   Lipid	Clinical chemistry/auto analyzer
   Protein	Clinical chemistry/auto analyzer
   Small molecule	Clinical chemistry/auto analyzer
   Steroid	Clinical chemistry/auto analyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Time at room temperature	0.5 h 1 h 2 h 4 h 24 h
   Storage	Freeze/thaw cycling	0 cycles 1 cycle 3 cycles 6 cycles 9 cycles
   Biospecimen Aliquots and Components	Blood and blood products	Serum Whole blood
   Biospecimen Preservation	Type of fixation/preservation	Frozen None (fresh)
   Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated

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