Detection of North American eastern and western equine encephalitis viruses by nucleic acid amplification assays.
We have developed nucleic acid sequence-based amplification (NASBA), standard reverse transcription PCR (RT-PCR), and TaqMan nucleic acid amplification assays for the rapid detection of North American eastern equine encephalitis (EEE) and western equine encephalitis (WEE) viral RNAs from samples collected in the field and clinical samples. The sensitivities of these assays have been compared to that of virus isolation. While all three types of nucleic acid amplification assays provide rapid detection of viral RNAs comparable to the isolation of viruses in Vero cells, the TaqMan assays for North American EEE and WEE viral RNAs are the most sensitive. We have shown these assays to be specific for North American EEE and WEE viral RNAs by testing geographically and temporally distinct strains of EEE and WEE viruses along with a battery of related and unrelated arthropodborne viruses. In addition, all three types of nucleic acid amplification assays have been used to detect North American EEE and WEE viral RNAs from mosquito and vertebrate tissue samples. The sensitivity, specificity, and rapidity of nucleic acid amplification demonstrate the usefulness of NASBA, standard RT-PCR, and TaqMan assays, in both research and diagnostic settings, to detect North American EEE and WEE viral RNAs.
Lambert
Amy J
AJ
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Fort Collins, Colorado 80521, USA. ahk7@cdc.gov
Martin
Denise A
DA
Lanciotti
Robert S
RS
D000818:Animals; D002522:Chlorocebus aethiops; D009033:Culicidae / Q000821:virology; D004663:Encephalitis Virus, Eastern Equine / Q000235:genetics / Q000302:isolation & purification*; D004667:Encephalitis Virus, Western Equine / Q000235:genetics / Q000302:isolation & purification*; D006736:Horses / Q000821:virology; D006801:Humans; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014709:Vero Cells
D012367:RNA, Viral