Discrimination of West Nile virus and Japanese encephalitis virus strains using RT-PCR RFLP analysis.
West Nile (WN) virus is a mosquito-borne flavivirus that induces lethal encephalitis in humans and horses. Since an outbreak of WN encephalitis in humans and horses occurred in New York City in late August 1999, the possibility exists that WN virus will invade regions that have close links with the United States, such as Japan. We developed a genetic diagnostic method that discriminates between strains of WN virus and Japanese encephalitis (JE) virus. The method involves RT-PCR restriction fragment length polymorphism (RFLP) analysis with a RT-PCR primer set, a nested PCR primer set, and a restriction enzyme. We detected WN and JE viruses in experimentally infected animal brain, spleen, and serum samples. Our method is useful in distinguishing WN viruses from the endemic background of JE viruses, and in discriminating the highly virulent WN strain, which was isolated in New York in 1999, from other WN virus strains.
Shirato
Kazuya
K
Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
Mizutani
Tetsuya
T
Kariwa
Hiroaki
H
Takashima
Ikuo
I
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000821:virology; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics*; D004196:Disease Outbreaks; D004664:Encephalitis Virus, Japanese / Q000235:genetics* / Q000302:isolation & purification; D004672:Encephalitis, Japanese / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009519:New York City / Q000453:epidemiology; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013154:Spleen / Q000821:virology; D014766:Viremia / Q000821:virology; D014774:Virulence; D014901:West Nile Fever / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D014902:West Nile virus / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity
D004279:DNA, Viral