Takeda Mayu, Funato Tadao, Ikemoto Masaki, Nanmoku Toru, Urasaki Yoshimasa, Iwatani Yoshinori
Department of Clinical Laboratory Science, Faculty of Health Care, Tenri Health Care University, Tenri, Japan; Department of Biomedical Informatics, Division of Health Sciences, Graduate School of Medicine, Osaka University, Suita, Japan.
J Clin Lab Anal. 2015 Jan;29(1):61-7. doi: 10.1002/jcla.21729. Epub 2014 Mar 28.
The collection of clinical samples, such as bone marrow (BM) and peripheral blood, is an important procedure for the extraction of the cellular RNA. It is essential to preserve the extracted RNA during and after the collection of clinical samples to ensure the accurate analysis of gene expression. To date, the PAXgene™ Blood RNA System has been proven useful for stabilizing RNA extracted from peripheral blood; however, a problem concerning the stability of the total RNA stored using the system has been identified. The PAXgene™ Bone Marrow RNA System (BM system) is a newly developed system, and its clinical usefulness as a stabilizer for the cellular RNA in BM and peripheral blood was investigated with respect to the quality of RNA extracted using this system. A quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was carried out using total RNA extracted with the BM system, which showed that total RNA was more stable in the BM system than in the conventional system, indicating that the BM system can be applied to RT-PCR. The BM system enabled us to detect Wilms' tumor suppressor gene (WT1) more effectively than the conventional system. In conclusion, the BM system is clinically valuable for extracting and stabilizing total RNA of high quality.
采集临床样本,如骨髓(BM)和外周血,是提取细胞RNA的重要步骤。在采集临床样本期间及之后保存提取的RNA对于确保基因表达的准确分析至关重要。迄今为止,PAXgene™血液RNA系统已被证明可用于稳定从外周血中提取的RNA;然而,已发现使用该系统储存的总RNA的稳定性存在问题。PAXgene™骨髓RNA系统(BM系统)是一种新开发的系统,针对使用该系统提取的RNA质量,研究了其作为BM和外周血中细胞RNA稳定剂的临床实用性。使用BM系统提取的总RNA进行了定量逆转录聚合酶链反应(RT-PCR),结果表明总RNA在BM系统中比在传统系统中更稳定,这表明BM系统可应用于RT-PCR。与传统系统相比,BM系统使我们能够更有效地检测威尔姆斯肿瘤抑制基因(WT1)。总之,BM系统在提取和稳定高质量总RNA方面具有临床价值。