Submit Manuscript  

Article Details


The Influence of Pre-analytical Factors on the Analysis of Circulating MicroRNA

[ Vol. 7 , Issue. 3 ]

Author(s):

Hiromichi Shiotsu, Kazuhiro Okada, Tatsuki Shibuta, Yuki Kobayashi, Saki Shirahama, Chieri Kuroki, Saori Ueda, Masanori Ohkuma, Katsuyoshi Ikeda, Yukio Ando, Hirotaka Matsui, Yuzo Kayamori and Tsukuru Umemura*   Pages 195 - 203 ( 9 )

Abstract:


Background: MicroRNAs (miRNA) are expected as useful biomarkers for various diseases. We studied the pre-analytical factors causing variation in the analysis of miRNA.

Material and Methods: Blood samples were collected from 25 healthy subjects. Plasma and serum were obtained from the same samples. The levels of miR-451, -16, -126, and -223 were analyzed using RT-qPCR. Cel-miR-39 was added as a spiked-in control in each sample.

Results: With the exception of miR-451, the levels of the miRNAs in plasma were higher than in serum. After high-speed centrifugation, the levels of miRNAs were almost equal between plasma and serum except for miR-451. Membrane filtration with 0.45 µm pore size reduced the levels of plasma miRNAs. The coagulation accelerators for serum processing did not affect the analysis of miRNA. The use of fraction containing particles of > 0.45 µm in size showed the inhibitory effect on the analysis of plasma miR-451. The RNase inhibitor was effective for protecting against the degradation of miRNAs.

Conclusion: Plasma contains factors modifying miRNA profiles. The immediate processing of plasma with membrane filtration and RNase inhibitor may be a relevant method for achieving the stable analysis of miRNA.

Keywords:

microRNA, plasma, serum, standardization, apoptosis, protien.

Affiliation:

Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Laboratory Medicine, Kumamoto University Hospital, Kumamoto, Department of Laboratory Medicine, Kumamoto University Hospital, Kumamoto, Department of Neurology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Department of Laboratory Medicine, Kumamoto University Hospital, Kumamoto, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka

Graphical Abstract:



Read Full-Text article