Abstract
BackgroundAdvances in microRNAs (miRNAs) biomarkers have generated disease markers with potential clinical values. However, none of these published results have been applied in clinic until today. The main reason could be the lack of simple but robust miRNA measurements.MethodsWe built up a simple but ultrasensitive RT-qPCR protocol, Direct S-Poly(T) Plus assay, for detecting miRNAs without RNA purification. In this study, the method was optimized and compared with other RNA purification-based miRNA assays, and the sensitivity was tested. Using Direct S-Poly(T) Plus method, seven potential miRNA biomarkers of colorectal cancer were validated.ResultsIt is possible to detect approximately 100 miRNAs with minimal plasma inputs (20 μl) and time (~ 140 min) with this approach. The sensitivity of this method was 2.7–343-fold higher than that of the stem-loop method, and comparable with S-Poly(T) plus method. 7 validated miRNA biomarkers of colorectal cancer by Direct S-Poly(T) plus assay could discriminate colorectal cancer stage I from healthy individuals, and promised satisfactory discrimination with the area under receiver operating characteristic (ROC) curve ranging from 0.79 to 0.94 (p value < 0.001).ConclusionsThis simple and robust protocol may have strong impact on the development of specific miRNAs as biomarkers in clinic.
Highlights
Advances in microRNAs biomarkers have generated disease markers with potential clinical values
Circulating miRNAs correlate with diagnosis, prognosis and responses to treatment [14,15,16]. These findings suggest that circulating miRNAs have great potential as biomarkers in monitoring the body’s physiopathology status
The general rules for this objective are the incorporation of tween 20 or proteinase K or high-temperature processing [28, 29]
Summary
Advances in microRNAs (miRNAs) biomarkers have generated disease markers with potential clinical values. None of these published results have been applied in clinic until today. Advances in miRNA biomarkers have generated some candidate markers of CRC with potential clinical values [21,22,23]. These published results have not been clinically applicable until today. The possible reasons could be the lack of sensitive and applied method in clinic, as well as knowledge about which biomarker(s) are stable and reproducible for clinical use
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