Objective: To develop a new nucleic acid sequence-based amplification (NASBA) method which is based on fluorescence probe for the detection of enterovirus and clinic diagnosis. Methods: The special primers and probes were designed based on the nucleic acid sequences of relative enterovirus in Gene bank. The commercial immunomagectic beads were conjugated with the primers and probes using nanotechnology. The conjugated products were amplified by NASBA and sequentially detected by NucliSens reader. The results were validated by real time-PCR. The sensitivity, repetition and specificity of the method were evaluated respectively. Results: The method could detect enterovirus with high specificity and stability as well as no interactions with other respiratory virus such as RSV. Conclusion: The new NASBA method is convenience, fast, sensitive and specific for enterovirus detection, which could be applied for regular surveillance and urgent investigation of outbreaks of enterovirus in the near future.