Detection of California serogroup viruses using universal primers and reverse transcription-polymerase chain reaction.
Universal primers have been identified and a protocol developed that are suitable for rapid detection of California encephalitis (CE) complex viruses in a reverse transcription-polymerase chain reaction (RT-PCR) assay. These primers correspond to sequences in the coding regions of the G2 glycoprotein of the middle-size RNA segment. The identities of the amplified products were confirmed by sequencing on the clones or PCR products. The technique is capable of detecting 40 plaque-forming units (PFU) directly on an ethidium bromide-stained agarose gel and the sensitivity increases to 0.4-1 PFU when a radiolabeled probe is used as the detector.
Campbell
W P
WP
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509, USA.
Huang
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D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D002141:Encephalitis Virus, California / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006023:Glycoproteins / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D012194:RNA-Directed DNA Polymerase; D012680:Sensitivity and Specificity; D012703:Serotyping
D017931:DNA Primers; D006023:Glycoproteins; D012333:RNA, Messenger; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase