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Muscovy duck parvovirus(MDPV)causes high mortality and morbidity in ducks.This study investigated a novel aptamer-based,label-free aptasensor detection of MDPV.In this study,we developed an ssDNA aptamer using the filtration partition and lambda exonuclease method with an affinity-based monitor and counter-screening process.After 15 rounds of SELEX(systematic evolution of ligands by exponential enrichment),the ssDNA aptamer Apt-10,which specifically bound to MDPV with high affinity(Kd=467 nM)was successfully screened,and the aptamer was also found to be good specific to MDPV.The selected Apt-10 aptamer can be used to distinguish MDPV and goose parvovirus(GPV).Three-dimensional structural analysis of the Apt-10 aptamer indicated that it folded into a compact stem-loop motif,which was related to its high affinity.Finally,a label-free detection method based on unmodified gold nanoparticles and Apt-10 aptamer was developed for MDPV determination.The concentration of Apt-10 aptamer at 5 μM was optimal for MDPV determination in the label-free aptasensor.Excellent linearity was acquired and the lowest detection limit was 1.5 or 3 EID50(50%egg infection dose)of MDPV,respectively,depending upon spectrophotometry or the naked eye were used.These results show the potential of the aptamer for the rapid detection of MDPV and antiviral research.