This protocol aims to amplify enterovirus (EV) A71 viruses but not other viruses.This protocol was designed by us.The oligonucleotides target the 5'UTR noncoding region. This is a qualitative test for investigating EV-A71 infection of humans.The test has identified both historical EV-A71 strains and contemporary strains and has confirmed these using subgenomic sequencing of partial 5'UTR-VP2 and partial VP1 sequencing. Culture of the virus is not required as this assay is capable of detecting EV-A71 direct from extracted clinical samples.