摘要
生物样本为转化医学研究提供了宝贵的临床资源.高效的生物样本质量检测技术对于临床样本分析结果的准确性和可靠性具有重要意义.将有效的质控检测方法和特定的生物学标志物作为血液质量指标,能够评估血液离体后的质量变化情况,进而在样本分析前剔除低质量样本,提升被分析样本和数据的总体质量和可靠性.血液样本由血细胞和血浆组成,包含核酸水平、蛋白质水平、代谢物水平等多个分子层面信息.因此在分析样本前,应根据样本类型和目标分子做出相应的质量评估.目前血细胞中的核酸质量可利用多种检测技术对其浓度、纯度和片段完整性进行检测.对于血浆和血清中的游离DNA以及结构不稳定的RNA小分子,可利用对应的靶标分子作为整体质量检测指标.但血细胞中mRNA离体表达水平的变化暂无明确的评估方法.此外,对于结构更为复杂的代谢小分子、蛋白质以及多肽片段,目前的研究多利用核磁共振技术或各种分离纯化手段(包括色谱、免疫亲和分离、磁分离等)与质谱联用技术来寻找目标质控靶标分子.这些分子作为标志物的可靠性、稳定性和准确性仍需验证.目前对于代谢小分子、蛋白质及多肽的质谱鉴定技术的成本高,无法满足大部分实验室对于样本质量检测的需求,因此需要寻找可靠的质量标志物、开发新的检测手段来降低血液样本质量评估的成本,顺利完成代谢小分子、蛋白质以及多肽片段的质量检测.
Biological samples play essential roles in biomedical studies, especially in "-omics" analyses.Recently, researchers have found that preanalytical effects and storage duration have direct effects on molecular biology-based procedures. The lack of guidelines for the collection, transport, and storage of samples could lead to the degradation of target molecules such as DNA, RNA, and proteins. These may also lead to inaccurate results in research laboratories. Therefore, sample quality should be assessed at the nucleic acid, metabolite, and protein levels depending on sample components, research objectives, and detection technologies. This review describes the specific biomarkers and proper tools for monitoring the quality of human blood in clinical laboratories.Genomic DNA(g DNA), messenger RNA(m RNA), cell-free DNA(cf DNA), and small noncoding RNA(mi RNA)are the main nucleic-acid components of blood samples used in research. This review summarizes the techniques used for the purification, yield analysis, and integrity analysis of g DNA and total RNA, including UV spectrophotometric analysis, qubit fluorometric quantification, and agarose gel electrophoresis. The internal reference genes used in cf DNA and mi RNA quality control are also listed. Quantitative real-time polymerase chain reaction analysis of reference genes is commonly conducted, and the results reflect the quantity and fragment integrity of cf DNA and mi RNA. However, no effective markers have been used for m RNA quality control. Therefore, the development of effective biomarkers is required for quantifying m RNA and small RNA molecules. Moreover, effective markers can be used for detecting changes in m RNA expression levels in vitro.Regarding metabolism, nuclear magnetic resonance(NMR) spectroscopy is the most common analytical method for the study of small metabolic molecules in blood. Matching between the available NMR data and reference databases can be applied to assess significant changes in metabolites and to evaluate blood sample quality. In
作者
孙青
梁锴
李岩
SUN Qing;LIANG Kai;LI Yan(Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China)
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2021年第8期938-946,共9页
Progress In Biochemistry and Biophysics
基金
中国科学院战略性先导科技专项基金(XDB38010000)
中国科学院战略生物资源能力建设项目(KFJ-BRP-017-01)资助
国家自然科学基金(21705160)。
