Carbon nanotubes and branched-DNA based nucleic acid assays: Towards a PCR-free detection and quantification of nucleic acids

Abstract

This research is directed towards the development of PCR-free ultrasensitive nucleic acids detection based on signal amplification approach. The solution holds promise for simple and low cost nucleic acids detection in portable devices, point-of-care and early disease diagnostic applications. Two types of nucleic acid assays based on novel carbon nanotube (CNT)-based label and branched-DNA (bDNA) amplifier were developed for the detection of a human acute lymphocytic leukaemia (ALL) oncogene, namely, p185 BCR-ABL fusion transcript. These assays have been successfully validated for PCR-free detection and quantification of p185 BCR-ABL in the samples of mRNA population extracted from positive ALL cell line SUP-B15, in comparison to those obtained from negative cell lines HL-60. Without the PCR amplification, the electrochemical nucleic acids assays utilizing CNT-based label and bDNA amplifier enabled direct detection of p185 BCR-ABL in as little as 65 ng and 4.6 ng of mRNA, respectively.