Simultaneous detection of dual single-base mutations by capillary electrophoresis using quantum dot-molecular beacon probe.
Here a novel capillary electrophoresis (CE) for simultaneous detection of dual single-base mutations using quantum dot-molecular beacon (QD-MB) probe is described. Two QD-MB probes were designed using 585 and 650-nm emitting CdTe QDs which were covalently conjugated to MBs with different DNA oligonucleotide sequences by amide linkage and streptavidin-biotin binding, respectively. The hybridizations of QD-MB probes with different DNA targets were then monitored by CE, and results indicated that the two QD-MB probes specifically hybridized with their complementary DNA sequences, respectively. Target DNA identification was observed to have a high sensitivity of 16.2 pg in CE. Furthermore, the simultaneous detection of dual single-base mutations in a given DNA oligonucleotide was successfully achieved in CE using above two QD-MB probes. This novel CE-assisted QD-MB biosensor offers a promising approach for simultaneous detection of multiple single-base mutations, and exhibits potential capability in the single nucleotide polymorphism (SNP) analysis and high-sensitivity DNA detection.
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, HuBei 430074, PR China.
Yong-Qiang
YQ
Li
Li-Yun
LY
Guan
Jian-Hao
JH
Wang
Hai-Li
HL
Zhang
Jun
J
Chen
Song
S
Lin
Wei
W
Chen
Yuan-Di
YD
Zhao
D020137:Base Pair Mismatch; D015374:Biosensing Techniques / Q000295:instrumentation; D004252:DNA Mutational Analysis / Q000295:instrumentation*; D015342:DNA Probes / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008856:Microscopy, Fluorescence / Q000295:instrumentation; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D045663:Quantum Dots; D017422:Sequence Analysis, DNA / Q000295:instrumentation*
D015342:DNA Probes