Reverse transcription-PCR detection of LaCrosse virus in mosquitoes and comparison with enzyme immunoassay and virus isolation.
A reverse transcription-PCR (RT-PCR) assay was developed and compared with enzyme immunoassay (EIA) and virus isolation for detecting LaCrosse virus (LAC) in mosquito pools. All three techniques were able to detect a single LAC-infected mosquito in a pool of 99 negative mosquitoes. Virus isolation was the most sensitive of the three techniques; it was possible to isolate virus immediately following intrathoracic inoculation of mosquitoes. RT-PCR was second in sensitivity; LAC RNA was detected 1 day postinfection. EIA detected LAC antigen 2 days postinfection. Additionally, RT-PCR and EIA were able to detect LAC RNA and protein, respectively, from mosquito samples which were subjected to seven freeze-thaw cycles, and RT-PCR was able to detect LAC RNA from mosquito samples which remained at room temperature for up to 7 days.
Wasieloski
L P
LP
Department of Microbiology, Colorado State University, Fort Collins 80523.
Rayms-Keller
A
A
Curtis
L A
LA
Blair
C D
CD
Beaty
B J
BJ
D000330:Aedes / Q000821:virology*; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D001483:Base Sequence; D005615:Freezing; D007124:Immunoenzyme Techniques; D018062:La Crosse virus / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013997:Time Factors; D014776:Virus Cultivation
D000956:Antigens, Viral; D012367:RNA, Viral