Nucleic acid amplification assays for detection of La Crosse virus RNA.
We report the development of nucleic acid sequence-based amplification (NASBA) and quantitative real-time reverse transcription (RT)-PCR assays for the detection of La Crosse (LAC) virus in field-collected vector mosquito samples and human clinical samples. The sensitivities of these assays were compared to that of a standard plaque assay in Vero cells. The NASBA and quantitative real-time RT-PCR assays demonstrated sensitivities greater than that of the standard plaque assay. The specificities of these assays were determined by testing a battery of reference strain viruses, including representative strains of LAC virus and other arthropod-borne viruses. Additionally, these assays were used to detect LAC viral RNA in mosquito pool samples and human brain tissue samples and yielded results within less than 4 h. The NASBA and quantitative real-time RT-PCR assays detect LAC viral RNA in a sensitive, specific, and rapid manner; these findings support the use of these assays in surveillance and diagnostic laboratory systems.
Lambert
Amy J
AJ
Division of Vector-Borne Infectious Diseases, National Centers for Infectious Diseases, CDC, Rampart Rd., Fort Collins, CO 80521, USA. ahk7@cdc.gov
Nasci
Roger S
RS
Cropp
Bruce C
BC
Martin
Denise A
DA
Rose
Becky C
BC
Russell
Brandy J
BJ
Lanciotti
Robert S
RS
D000818:Animals; D002522:Chlorocebus aethiops; D009033:Culicidae / Q000821:virology*; D004670:Encephalitis, California / Q000821:virology*; D006801:Humans; D018062:La Crosse virus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014709:Vero Cells; D010948:Viral Plaque Assay
D012367:RNA, Viral