Abstract: Provided herein are methods for determining the presence or absence of an enteroviruses and parechoviruses in a biological sample. The methods involve identifying the presence or absence of a target nucleic acids from the viruses using direct amplification from a biological sample without a step of extraction of the nucleic acids, but retaining substantially the same specificity and sensitivity of methods assaying extracted nucleic acids. Also provided are methods of diagnosis using the methods provided and compositions and kits for the practice of the methods.