Xinbing Zuo

Nucleic acids detection using cationic fluorescent polymer based on one-dimensional microfluidic beads array

An assay for rapid and direct detection of DNA/mRNA using cationic fluorescent polymer based on one-dimensional microfluidic beads array (1-D chip) has been developed. The cationic water-soluble polythiophene derivatives can easily transduce hybridization events into measurable optical signal due to the conformational changes of the conjugated backbone, when mixed with single-stranded or double-stranded oligonucleotides. In this paper, the polymer was introduced into 1-D chip for fluorescence detection of nucleic acids, and demonstrated its capability on rapid detection of p53 complementary DNA (cDNA) with different concentration. Using this system, we have evaluated the mRNA expression changes of three tumor-associated genes (p53, c-myc and cyclin-d1) in human nasopharyngeal carcinoma CNE2 cell lines before and after 5-flouorouracil (5-FU) stimuli. These results were validated by the conventional reverse transcriptase-PCR. The most important advantage of this assay is not needed target or report labeling prior to hybridization, which makes the experiment process easy to handle and low-cost for multi-target measurement.

One-Dimensional Microfluidic Beads Array for Nucleic Acids Detection

A one-dimensional addressable beads array for nucleic acids detection is presented in this paper. Molecular beacons (MBs) modified porous silica micro-beads were deposited along a micro-channel in sequence. This MB-functional beads array cooperating with microfluidic control allows multiple nucleic acid targets detection simultaneously with only once samples injection, a rapid dynamic hybridization and no need of samples labeling. As a demonstration, the synthesized DNA targets according to the same sequence with a section of human gene p53 mRNA was investigated with a limit of detection as few as 0.5 nM. We have further examined the asymmetry PCR (a-PCR) products of human gene p53, c-myc and cyclin-d1 to detect the mRNA expression changes before and after anticancer drug 5-flouorouracil (5-FU) treatment Such a platform used for nucleic acids detection has shown its advantages of handling simply, little samples consumption, low potential of contaminations, and low cost. Also, it have a potentially application to pursue high-throughput nuclei acids detection in a single channel.