Highly sensitive and specific multiplexed microRNA quantification using size-coded ligation chain reaction

Analytical Chemistry
Pengbo ZhangZhengping Li

Abstract

As important regulators of gene expression, microRNAs (miRNAs) are emerging as novel biomarkers with powerful predictive value in diagnosis and prognosis for several diseases, especially for cancers. There is a great demand for flexible multiplexed miRNA quantification methods that can quantify very low levels of miRNA targets with high specificity. For further analysis of miRNA signatures in biological samples, we describe here a highly sensitive and specific method to detect multiple miRNAs simultaneously in total RNA. First, we rationally design one of the DNA probes modified with two ribonucleotides, which can greatly improve the ligation efficiency of DNA probes templated by miRNAs. With the modified DNA probes, the ligation chain reaction (LCR) can be well applied to miRNA detection and as low as 0.2 fM miRNA can be accurately determined. High specificity to clearly discriminate a single nucleotide difference among miRNA sequences can also be achieved. By simply coding the DNA probes with different length of oligo (dA) for different miRNA targets, multiple miRNAs can be simultaneously detected in one LCR reaction. In our proof of principle work, we detect three miRNAs: let-7a, mir-92a, and mir-143, which can also be simul...Continue Reading

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Citations

Dec 29, 2015·ACS Applied Materials & Interfaces·Yuanyuan ZhouYongqiang Cheng
May 24, 2014·Chemical Communications : Chem Comm·Ya-ping ZengChun-yang Zhang
Mar 22, 2018·The Analyst·Yongqiang ChengZhengping Li
Jul 6, 2014·Chemistry : a European Journal·Federica DegliangeliRoberto Fiammengo
May 3, 2014·Angewandte Chemie·Susana CampuzanoJosé M Pingarrón
Jul 12, 2016·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Rui Tian, Xingwang Zheng
Dec 18, 2014·World Journal of Gastroenterology : WJG·Juan XieGuo-Qing Li
Apr 22, 2020·Organic & Biomolecular Chemistry·Pengbo ZhangZhengping Li
Dec 17, 2014·Analytical Chemistry·David W WegmanSergey N Krylov

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