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Published on: October 26, 2017
Centrifugation: an important pre-analytic procedure that influences plasma microRNA quantification during blood
Xiao-Hui Zheng1, Cui Cui, Xin-Xi Zhou
1State Key Laboratory of Oncology in South China, Bank of Tumor Resource, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong 510060, P. R. China. jiaweih@mail.sysu.edu.cn.
Chinese Journal of Cancer
|April 23, 2013
Summary
Two-step centrifugation significantly reduces plasma microRNA detection by pelleting microRNAs within large nanoparticles. This impacts microRNA biomarker studies, necessitating careful consideration of sample processing methods for accurate quantification.
Area of Science:
- Biochemistry
- Molecular Biology
- Biomarker Discovery
Background:
- Circulating microRNAs (miRNAs) are valuable biomarkers for cancer diagnosis and prognosis.
- Plasma/serum miRNA quantification is crucial for biomarker studies.
- Centrifugation is a standard method for preparing plasma/serum samples.
Purpose of the Study:
- To compare the effects of one-step and two-step centrifugation on plasma microRNA quantification.
- To investigate the underlying mechanisms affecting miRNA yield based on centrifugation method.
Main Methods:
- Plasma samples were processed using one-step and two-step centrifugation.
- MicroRNA quantification was performed using microarray and quantitative real-time PCR (Q-PCR).
- Dynamic light scattering was used to analyze nanoparticle size distribution.
Main Results:
- Two-step centrifugation significantly decreased microRNA signal detection rates compared to one-step.
- Recoverable microRNAs were found in the precipitate from the second centrifugation step.
- A reduction in nanoparticles >1,000 nm was observed in two-step processed plasma.
- Q-PCR confirmed a marked decrease in three specific microRNAs after two-step centrifugation.
Conclusions:
- Centrifugation method critically affects plasma microRNA yield.
- Exosomes or protein complexes containing miRNAs may form large nanoparticles, leading to their loss during two-step centrifugation.
- Standardization of sample processing is essential for reliable miRNA biomarker research.
