Use of the polymerase chain reaction for the sensitive detection of St. Louis encephalitis viral RNA

Daniel Keith Howe, Michael Harold Vodkin, Robert John Novak, Robert Ellis Shope, Gerald Lee McLaughlin

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


Polymerase chain reaction (PCR) assays were developed for the detection of RNA from the St. Louis encephalitis (SLE) virus. Using computer-assisted analysis of the MSI-7 strain SLE virus genome, two primer pairs were selected from the capsid-coding and the membrane associated protein-coding genes, and one from the envelope-coding gene. Reverse transcription was primed with either specific oligomers or with random hexamers; these methods were compared for cDNA synthesis and subsequent PCR amplification with the oligomeric pairs. Random hexamers provided more sensitive detection of viral RNA. Each primer pair specifically amplified the expected sized fragment from the Parton SLE strain grown in Aedes albopictus cells, but did not amplify Aedes albopictus cell RNA controls. The technique also detected SLE virus RNA in 1 pg of total cellular RNA added to a background of 1 μg boiled brain tissue, and in 0.5 pg of total RNA added to homogenized mosquito abdomen. PCR-based assays may be adaptable to detect SLE virus RNA in naturally infected mosquitoes, birds, and human cerebrospinal fluid and brain.

Original languageEnglish
Pages (from-to)101-110
Number of pages10
JournalJournal of Virological Methods
Issue number1
StatePublished - Jan 1992

Bibliographical note

Funding Information:
This work was supported in part by funds from WHO TDR and PHS ROl EY082905 (GLM), and SENR TM-5 (RJN), PHS AI10984 and U.S. Army DAMD17-90-Z-0020 (RES). Computer support was provided to MHV by the Pittsburgh Supercomputer Center grant PSCB DMB890077P. We thank Drs. B. Fonseca and P. Mason from the Yale Arbovirus Research Unit for advice and assistance.


  • Flavivirus
  • Polymerase chain reaction
  • St. Louis encephalitis virus

ASJC Scopus subject areas

  • Virology


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