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Research Article Open Access
Background: MicroRNA (miRNA) molecules have been detected in many body fluids and used as biomarkers. However, blood cell-derived miRNA molecules due to hemolysis have been shown to affect the amounts of plasma miRNAs. It is important to check the quality of plasma samples for clinical applications. In the present study, an objective method for quality checking plasma samples was investigated using permitted color tone level and the absorbance at 414 nm.
Material and method: An ROC analysis of the macroscopic color tone and the absorbance in 1213 clinical samples was performed. The optimal cut-off absorbance value was validated using the amount of plasma miRNA.
Results: AUC for detecting hemolyzed samples was very high (0.986) at a cut-off absorbance value of 1.664. A sensitivity and specificity were 99% and 92%, respectively. For validation study, 3 candidate miRNAs were selected by a miRNA microarray between fresh and hemolyzed plasma samples; the amounts of miR-16 and miR-19b increased in hemolyzed samples, whereas that of miR-223 remained unchanged. The amounts of plasma miR-16 and miR-19b were significantly increased in 5 clinical samples with higher absorbance values (p=0.0001 and p=0.0003, respectively), while the amounts of these miRNAs were not increased in samples with lower absorbance values.
Conclusions: A simple method for quality checking plasma samples using color tone and absorbance is very useful and significant.
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Author(s): Katsutoshi Shoda, Hirotaka Konishi, Daisuke Ichikawa, Yuji Fujita, Hidekazu Hiramoto, Junichi Hamada, Tomohiro Arita, Toshiyuki Kosuga, Shuhei Komatsu, Atsushi Shiozaki, Kazuma Okamoto and Eigo Otsuji
Plasma sample, Quality check, Color tone, Absorbance, miRNA, Biochemical pharmacology