Molecular beacon-based fluorescence biosensor for the detection of gene fragment and PCR amplification products related to chronic myelogenous leukemia.
A novel fluorescence method has been established for the determination of gene fragment and PCR amplification products related to chronic myelogenous leukemia (CML). A molecular beacon (MB) which comprises a DNA loop section, a pair of fluorophore (tetramethoxyl rhodamine, TAMRA), and a quencher (4-(2-methyl-on-amino-azobenzene) benzoate, DABCYL) was designed. The loop sequence of MB was designed according to the DNA sequence relating to CML (type b3a2) which contained a single-stranded oligonucleotide. Before hybridization, the fluorescence from the TAMRA had been quenched by the DABCYL. After hybridization with the complementary DNA, the quencher will become far away from the TAMRA, and the fluorescence intensity detected will increase. Changes in the fluorescence intensity have a linear relationship with the concentration of complementary DNA (C) in the range of 4.0 × 10(-9)-3.2 × 10(-8) mol/L, with a correlation coefficient of 0.9973; the detection limit was 6.0 × 10(-10) mol/L (S/N = 3). The developed method has high selectivity, which can be used to discriminate single-base mismatch sequence. The method has been applied to detect the short-stranded CML DNA fragment (278 bp) with high sensitivity. This approach is a promising method for the detection of CML in real samples for medical diagnostics.
Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou, China.
Ailin
A
Liu
Zhouliang
Z
Sun
Kun
K
Wang
Xuhai
X
Chen
Xiongwei
X
Xu
Yong
Y
Wu
Xinhua
X
Lin
Yuanzhong
Y
Chen
Min
M
Du
D015374:Biosensing Techniques / Q000379:methods*; D002138:Calibration; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics*; D005453:Fluorescence; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013696:Temperature
D004273:DNA, Neoplasm