title: MicroRNA-derived fragment length polymorphism assay.
abstract: MicroRNA (miRNA) studies are experiencing a transition from basic research applications to clinical applications. However, the lack of reliable and sensitive miRNA detection methods has become a bottleneck in the process. Here, we report an absolute quantification method based on the competitive PCR amplification of specific miRNAs and synthetic RNA spike-ins in a single reaction. RNA spike-ins are quantified as dynamic RNA copy number standards and are used to measure selected miRNAs free from the effects of intra-assay variables, including those from individual sample sources. Combined with the size differentiation power of capillary electrophoresis, the content of miRNAs was reproducibly measured, with verifiable detection limits of 10-46 copies over 5-log detection ranges. The direct measurements of miRNAs from 168 human serum samples and their considerable value as a diagnostic for bronchopneumonia and bronchiolitis demonstrate the potential of the assay in clinical applications.
authors: Dujiangyan Medical Center, Chengdu, Sichuan, 611830, China.
authors: Xiaoping
authors: X
authors: Xie
authors: Chengdu Nuoen Biotechnologies, Inc., Chengdu, Sichuan, 610041, China.
authors: Fang
authors: F
authors: Tang
authors: Dujiangyan Medical Center, Chengdu, Sichuan, 611830, China.
authors: Zhao
authors: Z
authors: Yang
authors: Chengdu Nuoen Biotechnologies, Inc., Chengdu, Sichuan, 610041, China.
authors: Yaoyi
authors: Y
authors: Zhang
authors: Chengdu Nuoen Biotechnologies, Inc., Chengdu, Sichuan, 610041, China.
authors: Zihao
authors: Z
authors: Feng
authors: Chengdu Nuoen Biotechnologies, Inc., Chengdu, Sichuan, 610041, China.
authors: Yu
authors: Y
authors: Yang
authors: Chengdu Nuoen Biotechnologies, Inc., Chengdu, Sichuan, 610041, China.
authors: Xiujin
authors: X
authors: Wu
authors: Chengdu Nuoen Biotechnologies, Inc., Chengdu, Sichuan, 610041, China.
authors: Feifei
authors: F
authors: Zhang
authors: Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, 610075, China.
authors: Jie
authors: J
authors: Zhu
authors: Chengdu Nuoen Biotechnologies, Inc., Chengdu, Sichuan, 610041, China.
authors: Kai
authors: K
authors: Xu
mesh_terms: D019075:Electrophoresis, Capillary; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results
chemical_list: D035683:MicroRNAs