A novel sandwich assay with molecular beacon as report probe for nucleic acids detection on one-dimensional microfluidic beads array.
A novel sandwich assay with molecular beacons as report probes has been developed and integrated into one-dimensional microfluidic beads array (1-D chip) to pursue a label-free and elution-free detection of DNA/mRNA targets. In contrast with the immobilized molecular beacons, this sandwich assay can offer lower fluorescence background and correspondingly higher sensitivity. Furthermore, this sandwich assay on 1-D chip operating in conjunction with molecular beacon technique allows multiple targets detection without the need of laborious and time-consuming elution, which makes the experiment process simple, easy to handle, and reproducible results. In the experiment, the synthesized DNA targets with different concentrations were detected with a detection limit of approximately 0.05 nM. Moreover, the mRNA expression changes in A549 cells before and after anticancer drug 5-flouorouracil treatments were detected and the results were validated by the conventional RT-PCR method.
Zuo
Xinbing
X
State Key Laboratory of Chemo/Biosensing & Chemometrics, Biomedical Engineering Center, College of Chemistry & Chemical Engineering, Hunan University, Engineering Research Center for Bio-Nanotechnology of Hunan Province, Changsha 410082, China.
Yang
Xiaohai
X
Wang
Kemin
K
Tan
Weihong
W
Wen
Jianhui
J
D001681:Biological Assay; D045744:Cell Line, Tumor; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D004247:DNA / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor; D005472:Fluorouracil / Q000627:therapeutic use; D006801:Humans; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics; D008863:Microspheres; D015335:Molecular Probes / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors
D015335:Molecular Probes; D012333:RNA, Messenger; D004247:DNA; D005472:Fluorouracil