MicroRNA-derived fragment length polymorphism assay

Scientific Reports
Xiaoping XieKai Xu

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.

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Citations

Sep 16, 2016·Chembiochem : a European Journal of Chemical Biology·Ashley R ConnollyMatt Trau
Jan 15, 2020·Signal Transduction and Targeted Therapy·Juanjuan KangJian Huang
Apr 17, 2020·Signal Transduction and Targeted Therapy·Juanjuan KangJian Huang

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Methods Mentioned

BETA
PCR
electrophoresis
Assay
blood collection

Software Mentioned

Excel
SPSS
miRbase
SPSS ( Statistical Package for the Social Sciences )
mfold

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