Pre-Analytical Modification of Serum miRNAs: Diagnostic Reliability of Serum miRNAs in Hemolytic Diseases

Journal of Clinical Medicine

Tetsuhiro Harada¹  Ayaka Ishibashi¹  Mari Koga¹  Kamonlak Leecharoenkiat²  Kenichi Sato³  Mayu Hatano³  Tatsuki Shibuta³  Yukichi Takada³  Tsukuru Umemura³  Hanae Shimura³  Supat Chamnanchanunt⁴  Takashi Hisatomi⁵  Noriyasu Fukushima⁶  Suthat Fucharoen⁷  Saovaros Svasti⁷ 

[1] Clinical Laboratory, Kouhoukai Takagi Hospital, Okawa 831-8501, Japan;Department of Clinical Microscope, Faculty of Medical Sciences, Chulalongkorn University, Bangkok 10330, Thailand;Department of Medical Technology and Sciences, International University of Health and Welfare, Okawa 831-8501, Japan;Faculty of Tropical Medicine, Mahidol University, Bangkok 100400, Thailand;Hematology, Kouhoukai Takagi Hospital, Okawa 831-8501, Japan;Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan;Thalassemia Research Center, Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhon Pathom 73130, Thailand;

关键词: microRNA;    miR-451a;    pre-analytical;    blood drawing;    serum separation;    hemolytic disease;   

DOI  :  10.3390/jcm10215045

来源: DOAJ

【 摘 要 】

Circulating microRNAs (miRNAs) are useful biomarkers of hemolysis. Since blood cells are the main origins of circulating miRNAs, we evaluated blood cell-related pre-analytical modification of the miRNA signatures during blood drawing and serum processing. The levels of miRNA before and after ex vivo blood drawing were analyzed with the reverse transcriptase-based polymerase chain reaction method. Furthermore, the changes of miRNA signatures caused by different time-lag between blood drawing and serum preparation by 24 h were evaluated. Finally, we compared the miRNA levels between leftover samples and samples of hemolytic diseases. Blood drawing procedure induced increments of red blood cell (RBC)-related miRNAs (miR-451a, miR-486) about 2-fold. One hour standing of blood samples before serum separation induced almost the same increases in RBC-related miRNAs. To test the clinical usefulness of miR-451a as a biomarker of hemolytic diseases, we analyzed miRNAs of samples from 10 normal subjects, 30 leftover samples in the clinical laboratory, and 20 samples from patients with hemolytic diseases. Serum miR-451a significantly increased in patients with hemolytic anemia more than the levels of pre-analytical modification. In conclusion, the pre-analytical modification of serum miRNAs did not disturb the usefulness of RBC-derived miRNAs as biomarkers of hemolytic diseases.

【 授权许可】

Unknown