Keywords: Surface Plasmon Resonance, miRNA, gold nanoparticles.

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            MircoRNAs (miRNA) are short single stranded molecules which control genes expression by degradation of mRNA or by inhibition of translation. The Role of miRNA in many pathological states makes them a potential candidate for biomarkers. Despite many research results that sustain the correlation between the changes of expression of miRNA and the progress of pathological state, there are many challenges that have to be overcome for the development of method for miRNA detection. The most characteristic feature of miRNA is their short length and high sequence similarity. Those factors may cause many problems in hybridization based detection methods. For this reasons a development of new methods of detection that are sensitive and reliable is essential.

            Surface Plasmon Resonance (SPR) based diagnostic methods are characterized by short time needed for obtaining an result, are sensitive, and don’t require labeling of molecules. In this work we present a new method for detection based on SPR in which after the hybridization of miRNA on the sensor surface with the PNA probe the 3’ and 5’ ends or miRNA are enzymatically and chemically modified with the use of poly(A) tail or biotin. After the functionalization of miRNA the detection stage is carried out by gold nanoparticles functionalized with poly(T) or streptavidin.

The analysis shows that the use of gold nanoparticles that enhance the SPR signal and specific PNA probes, it is possible to detect miRNA in subnanomolar concentrations. The results show that the sensitivity and versatility of this method of detection can be used for the development of diagnostic methods that don’t require amplification of genetic material.

Acknowledgments: the PhD scholarship is founded by The Foundation for Polish Science

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