Specific detection of echoviruses 22 and 23 in cell culture supernatants by RT-PCR.
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.
Oberste
M S
MS
Respiratory and Enteric Viruses Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA. mbo2@cdc.gov
Maher
K
K
Pallansch
M A
MA
D002478:Cells, Cultured; D003470:Culture Media; D017931:DNA Primers; D004457:Echovirus Infections / Q000821:virology; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification; D029822:Enterovirus B, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping; D013045:Species Specificity; D014779:Virus Replication
D003470:Culture Media; D017931:DNA Primers