Rapid detection of west nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay.
The authors report on the development and application of a rapid TaqMan assay for the detection of West Nile (WN) virus in a variety of human clinical specimens and field-collected specimens. Oligonucleotide primers and FAM- and TAMRA-labeled WN virus-specific probes were designed by using the nucleotide sequence of the New York 1999 WN virus isolate. The TaqMan assay was compared to a traditional reverse transcriptase (RT)-PCR assay and to virus isolation in Vero cells with a large number ( approximately 500) of specimens obtained from humans (serum, cerebrospinal fluid, and brain tissue), field-collected mosquitoes, and avian tissue samples. The TaqMan assay was specific for WN virus and demonstrated a greater sensitivity than the traditional RT-PCR method and correctly identified WN virus in 100% of the culture-positive mosquito pools and 98% of the culture-positive avian tissue samples. The assay should be of utility in the diagnostic laboratory to complement existing human diagnostic testing and as a tool to conduct WN virus surveillance in the United States.
Lanciotti
R S
RS
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado 80521, USA.
Kerst
A J
AJ
Nasci
R S
RS
Godsey
M S
MS
Mitchell
C J
CJ
Savage
H M
HM
Komar
N
N
Panella
N A
NA
Allen
B C
BC
Volpe
K E
KE
Davis
B S
BS
Roehrig
J T
JT
D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D001717:Birds / Q000821:virology; D001921:Brain / Q000821:virology; D002522:Chlorocebus aethiops; D009033:Culicidae / Q000821:virology*; D006801:Humans; D012367:RNA, Viral / Q000097:blood / Q000134:cerebrospinal fluid; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000378:metabolism*; D014709:Vero Cells; D014776:Virus Cultivation; D014901:West Nile Fever / Q000175:diagnosis* / Q000662:veterinary / Q000821:virology; D014902:West Nile virus / Q000235:genetics / Q000302:isolation & purification*
D012367:RNA, Viral; D019914:Taq Polymerase