Detection of enteroviral infection in paraffin-embedded tissue by the RNA polymerase chain reaction technique.
A method using the RNA polymerase chain reaction technique to diagnose infection retrospectively using single-stranded RNA enteroviruses in paraffin-embedded tissue blocks is reported. This method takes advantage of extreme sequence conservation in the 5' untranslated region of the enteroviral genome and uses two rounds of amplification with nested oligonucleotide primers, thus allowing rapid diagnosis without the use of radioactive reagents. The technique should prove useful in cases in which viral infection is suspected after histopathologic evaluation, when fresh or frozen tissues often are unavailable. The sensitivity of the method is demonstrated by successful amplification of Enterovirus 11 RNA extracted from 4-year-old liver tissue obtained at autopsy examination that was initially fixed and embedded 45 hours after death.
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Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts.
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Tycko
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D001483:Base Sequence; D004770:Enterovirus / Q000235:genetics* / Q000302:isolation & purification; D004769:Enterovirus Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000235:genetics; D005784:Gene Amplification; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000382:microbiology; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics*; D012189:Retrospective Studies
D009841:Oligonucleotides; D012367:RNA, Viral