Abstract
A high sensitive and selective method for sequence-specific DNA detection is developed using a non-labeled molecular beacon (MB) and a nucleic acid dye SYBR Green I. It is demonstrated by an oligonucleotide sequence of wild-type hepatitis B virus as a model system. Under the optimum conditions, the fluorescence intensity of SYBR Green I exhibit good linear dependence on quantity of target DNA in the range of 4 × 10<sup>-11</sup>-400 × 10<sup>-11</sup> mol/L. The fitted regression equation is DI = 1.9556<italic>C</italic> + 31.4659 with a correlation coefficient of 0.9956 (<italic>R</italic><sup>2</sup>), and the detection limit is 2 × 10<sup>-11</sup> mol/L (3<italic>s</italic>). The proposed method has a good precision, simple operation, fast detection speed, low detection limit, high accuracy and high sensitivity. The quantitative detection of hepatitis B virus is realized by this proposed method based on asymmetric PCR technology.
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