Quantitative analysis of La Crosse virus transcription and replication in cell cultures and mosquitoes.
نویسندگان
چکیده
La Crosse (LAC) virus (family Bunyaviridae, genus Orthobunyavirus) small (S) segment negative-sense RNA genome (vRNA), positive-sense full-length RNA complement (vcRNA), and subgenomic mRNA were assayed in infected cell cultures and female Aedes (Ochlerotatus) triseriatus mosquito tissues using quantitative PCR (Q-PCR). During persistent infection of C6/36 (Aedes albopictus) and MAT (Aedes triseriatus) cultured cells and cytolytic infection of BHK-21 cultured cells, LAC vRNA was the most abundant RNA species, followed by mRNA and vcRNA. RNA copy numbers per cell were quantified and vRNA correlated to virus titer in cell culture medium. The Q-PCR assay proved more sensitive than reverse transcription (RT)-PCR and immunofluorescence assays (IFA) for detecting LAC virus infection of mosquitoes. After infection of female mosquitoes orally, quantities of LAC RNA increased in ovaries for 6 days, and as ovarian biosynthetic activity quiesced, LAC RNA quantities decreased then remained detectable at a low level. After a second, noninfectious blood meal, quantities of LAC RNA in ovaries increased significantly, quantitatively confirming correlation of LAC virus RNA synthesis with vector metabolic activity. Coregulation of viral replication and mosquito ovary metabolic activity may condition efficient transovarial transmission.
منابع مشابه
Modulation of La Crosse Virus Infection in Aedes albopictus Mosquitoes Following Larval Exposure to Coffee Extracts
The mosquito-borne La Crosse virus (LACV; Family Bunyaviridae) may cause encephalitis, primarily in children, and is distributed throughout much of the eastern United States. No antivirals or vaccines are available for LACV, or most other mosquito-borne viruses, and prevention generally relies on mosquito control. We sought to determine whether coffee extracts could interfere with LACV replicat...
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ورودعنوان ژورنال:
- The American journal of tropical medicine and hygiene
دوره 74 2 شماره
صفحات -
تاریخ انتشار 2006