Microgenomics 2021 webinar EV & Circulating Biomarkers: MKubista
Channel: The International Symposium on Microgenomics 2023
Date: 2022-01-03

Mikael Kubista (Institute for Biotechnology, Republic Czech) presents "Two-tailed PCR for Precision Diagnostics"

We present a highly specific, sensitive and cost-effective system to quantify miRNA and cell-free DNA in liquid biopsies based on novel chemistry called Two-tailed PCR. Two-tailed PCR takes advantage of target-specific primers composed of two hemiprobes complementary to two different parts of the target molecule connected by a hairpin structure. The introduction of a short hemiprobe that senses the variable sequences confers exceeding sequence specificity while maintaining the very high sensitivity of PCR. Highly similar targets can be distinguished with superior precision irrespectively of the position of the mismatched nucleotide. Further, the target molecule can be very short, making Two-tailed PCR the preferred method for microRNA profiling as well as analysis of rare sequence variants in cell-free DNA or RNA and formalin fixed paraffine embedded (FFPE) tissue. Two-tailed RT-qPCR has a dynamic range of 7 logs and a sensitivity to detect less than ten target miRNA molecules. Two-tailed PCR is readily multiplexed.

(1) Two-tailed RT-qPCR: a novel method for highly accurate miRNA quantification. P Androvic, L Valihrach, J Elling, R Sjoback, M Kubista. Nucleic acids research 45 (15), e144-e144