Exiqon LNA™ microRNA qPCR – Liquid Biopsies Revealed
Our understanding of RNA diversity and complexity is rapidly expanding. Particularly microRNAs have emerged as informative biomarkers of cell status and function. Sensitive, specific and unbiased microRNA profiling in liquid biopsies – from serum and plasma, to urine and CSF – holds great promise for diagnostics and disease monitoring as a non-invasive and relatively inexpensive analysis method.
The complexity of liquid biopsies make them difficult samples to analyse. They often have low microRNA content and harbour inhibitors of PCR. Commonly overlooked is the effect of collection, cellular contamination, and storage conditions, which can impact analysis outcome.
Nevertheless, with the right know-how and products designed to meet challenges in characterizing microRNA in liquid biopsies, it is possible to detect subtle differences in microRNA expression patterns that provide more refined insights into different health stages.
Building on decades of experience in pioneering technologies for microRNA research, Exiqon offers unique products and services that accurately reveal microRNAs in liquid biopsies. With proprietary LNA™ technology, the miRCURY™ LNA™ miRNA PCR System takes your qPCR experiments to the next level:
- Unparalleled sensitivity – reliably quantify individual microRNAs from as little as 1 pg total RNA.
- Engineered specificity – LNA™-enhanced primers are shorter, so both primers fit on the target microRNA without overlap to reveal single-nucleotide differences.
- Uniform coverage – LNA™-enhanced primers can be Tm-normalized for uniform detection of microRNA, regardless of target GC content.
- From RNA to data in 3 hours – Streamlined protocols with only two hands-on steps ensure high accuracy and throughput.
- Custom panels and assays – use pre-validated assays or intelligent primer design algorithms to build panels in 96- or 384-well plates that require just 20 ng total RNA.
The miRCURY™ LNA™ miRNA PCR System is perfectly suited for liquid biopsies. Developed to meet the stringent demands of biofluid biomarker discovery, the platform merges extreme sensitivity and specificity afforded by LNA™-enhanced primers with a universal reverse transcription step without need for pre-amplification. The system accurately and quickly profiles low-abundance microRNA species in liquid biopsies.
A powerful NGS validation tool
In a benchmarking study (2014, Nature Methods 11:809–815), Mestdagh et al. compared 12 quantitative microRNA expression platforms. The miRCURY™ LNA™ miRNA PCR System outperformed others, offering the best combination of sensitivity, specificity, accuracy and reproducibility.
Furthermore, 85-90% of differentially expressed microRNAs detected by NGS could be validated with our qPCR platform. In fact, our qPCR system was the only tested platform to cluster nicely with Illumina® and Ion Torrent® sequencing results.
Accessing information from challenging samples
microRNA detection in very dilute samples (e.g., urine, CSF) can be boosted by first isolating exosomes in the sample and then profiling microRNAs contained in these extracellular vesicles. The gold standard for exosome isolation is differential centrifugation, but the low throughput of this method is incompatible with larger sample numbers. The miRCURY™ Exosome Isolation Kits for serum, plasma, urine, CSF and cell culture supernatant enable efficient, highly standardized exosome isolation in less than 2 hours. Isolated exosomes exhibit similar size distribution but higher recovery rate compared to ultracentrifugation, and preserve microRNAs.
Standardized pre-analytics are key
Over the years, we have learned the importance of standardized and controlled sample collection and specimen pre-treatment protocols. Our monitoring and control technologies assess sample quality, including cellular contamination and hemolysis. We are happy to share our insights about pre-analytical variables and help you plan protocols that maximize the outcome of research projects with liquid biopsies.
Tackle liquid biopsies with sensitive and accurate qPCR and gain new insights. Contact us to learn more about LNA™-enhanced products or to discover our expert qPCR Services.