Broad-spectrum anti-SARS-CoV-2 strategies that can inhibit the infection of wild-type and mutant strains would alleviate their threats to global public health. Here, we propose an icosahedral DNA framework for the assembly of spatially ar-range up to thirty neutralizing aptamers (IDNA-30) to inhibit viral infection. Each triangular plane of IDNA-30 is com-posed of three precisely positioned aptamers topologically matching SARS-CoV-2 spike trimer, thus forming a multivalent spatially patterned binding. Due to its multiple binding sites and moderate size, multifaced IDNA-30 induces aggregation of viruses. The rigid icosahedron framework afforded by four-helixes not only forms a steric barrier to prevent the virus from binding to host, but also limits the conformational transformation of SARS-CoV-2 spike trimer. Combining multiva-lent topologically patterned aptamers with structurally well-defined nano-formulations, IDNA-30 exhibits excellent and broad-spectrum neutralization against SARS-CoV-2, including almost completely blocking the infection of Omicron pseu-dovirus. Overall, this multi-dimensional neutralizing strategy provides a new direction for the assembly of neutralizing reagents to enhance the inhibitory effect of SARS-CoV-2 infection and combat other disease-causing viruses.