Fast duplex one-step RT-PCR for rapid differential diagnosis of entero- or toscana virus meningitis

Acute meningitis is the most common neurologic disease that involves the central nervous system. The spectrum of infectious agents that cause neurologic infection is remarkably broad and numerous viruses are the most frequent cause of the aseptic meningitis syndrome. We applied a multiplex one-step method for the rapid detection of the genomic RNA of different neurotropic viruses: particles in the genus Enterovirus and Toscana virus, which are the most representative aetiologic agents in our country during the spring-summer period. We have evaluated the sensitivity and the specificity of the multiplex one-step test on positive controls and on RNA extracted from clinical samples harvested from 475 patients with meningitis hospitalized during the 1996-2001 period. The multiplex one-step RT-nPCR protocol allows for the detection of enterovirus and Toscana virus RNA in a single sample, by using, at the same time, a very small clinical sample volume. In our study we were able to diagnose 192 cases of meningitis by Toscana virus and 31 cases by enteroviruses out of 475 cases of meningitis utilizing the described one-step multiplex method.