The present disclosure provides methods that permit detection of all known species of Parechovirus, including Human parechovirus and Ljungan virus (LV). In particular examples the method includes amplifying at least a portion of the 5′NTR of parechovirus nucleic acid molecules obtained from a sample, and detecting the resulting amplicons, but does not require culturing of the virus. The present disclosure also provides methods for determining which particular species or serotype of parechovirus is present in a biological sample. Also provided are oligonucleotide primers and probes that can be used in the disclosed methods.