Specific detection of echoviruses 22 and 23 in cell culture supernatants by RT-PCR

Abstract

Reverse transcription–polymerase chain reaction (RT-PCR) methods are available for the rapid detection of enteroviruses in clinical specimens or virus isolates. Pan-enterovirus PCR primers, however, fail to amplify echovirus (E) type 22 or 23 because of their extreme sequence divergence from the other enteroviruses. We have developed an RT-PCR method to detect specifically E22 and E23 RNA directly in tissue culture supernatants without a viral RNA purification step. The E22/E23 primers successfully amplified 20 of 20 clinical isolates of E22 and 4 of 4 E23 isolates representing viruses isolated in 15 states over a 19-year period, as well as E22 and E23 prototype strains isolated in the 1950s. The primers did not amplify any of the other 64 enterovirus prototype strains. J. Med. Virol. 58:178–181, 1999. Published 1999 Wiley-Liss, Inc.