REVERSE TRANSCRIPTION MULTIPLEX PCR FOR DIFFERENTIATION BETWEEN POLIOVIRUSES AND ENTEROVIRUSES FROM CLINICAL AND ENVIRONMENTAL-SAMPLES

For the rapid detection of polioviruses and their differentiation from nonpoliovirus enteroviruses, we developed a protocol in which clinica l or environmental specimens are first inoculated onto cell cultures i n tubes, After overnight incubation, the cultures are subjected to rev erse transcription multiplex PCR with a primer pair which detects all enteroviruses (T, Hyypia, P, Auvinen, and M. Maaronen, J. Gen, Virol, 70:3261-3268, 1989) and two newly designed primer pairs specific for a ll 36 poliovirus strains tested, The PCR products can unequivocally be identified by their lengths in agarose gels, whereas the genetic hete rogeneity of the poliovirus strains precludes identification by back-h ybridization with internal probes, The proposed protocol is highly ins ensitive to the inhibitory effects of substances in the sample (stool, sewage). It allows for the detection of polioviruses and for poliovir uses to be distinguished from nonpoliovirus enteroviruses within 24 h, and it allows for the concomitant isolation of a viable strain suitab le for further typing.