Primer-EV-forward coupled with primer-EV-reverse were designed as universal primers of enterovirus and used to amplify a 154-bp segment shared by enteroviruses,
including EV71, EV70, coxsackievirus A, and poliovirus. Primer-EV71-forward and -reverse were designed to detect EV71, whereas pCA16-
forward and -reverse were for CoxA16 detection. Primer EV71-VP1-forward and -reverse were used to construct pET28a-VP1.