1921
Volume 74, Issue 2
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

La Crosse (LAC) virus (family Bunyaviridae, genus ) 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 mosquito tissues using quantitative PCR (Q-PCR). During persistent infection of C6/36 () and MAT () 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.

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2006-02-01
2017-11-20
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References

  1. Calisher CH, 1994. Medically important arboviruses of the United States and Canada. Clin Microbiol Rev 7 : 89–116.
  2. Borucki MK, Kempf BJ, Blitvich BJ, Blair CD, Beaty BJ, 2002. La Crosse virus: replication in vertebrate and invertebrate hosts. Microbes Infect 4 : 341–350.
  3. Schmaljohn CS, Hooper J, 2001. Bunyaviridae: The viruses and their replication. Knipe DM, Howley P, eds. Virology. Vol. II. 4th Edition. Philadelphia: Lippincott Williams & Wilkins, 1581–1602.
  4. Chandler LJ, Wasieloski LP, Blair CD, Beaty BJ, 1996. Analysis of La Crosse virus S-segment RNA and its positive-sense transcripts in persistently infected mosquito tissues. J Virol 70 : 8972–8976.
  5. Drosten C, Gottig S, Schilling S, Asper M, Panning M, Schmitz H, Gunther S, 2002. Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR. J Clin Microbiol 40 : 2323–2330.
  6. Garcia S, Crance JM, Billecocq A, Peinnequin A, Jouan A, Bouloy M, Garin M, 2001. Quantitative real-time PCR detection of Rift Valley fever virus and its application to evaluation of antiviral compounds. J Clin Microbiol 39 : 4456–4461.
  7. Boonham N, Smith P, Walsh K, Tame J, Morris J, Spence N, Bennison J, Barker I, 2002. The detection of Tomato spotted wilt virus (TSWV) in individual thrips using real time fluorescent RT-PCR (TaqMan). J Virol Methods 101 : 37–48.
  8. Roberts CA, Dietzgen RG, Heelan LA, Maclean DJ, 2000. Real-time RT-PCR fluorescent detection of tomato spotted wilt virus. J Virol Meth 81 : 1–8.
  9. Hutchinson KL, Rollin PE, Peters CJ, 1998. Pathogenesis of a North American hantavirus, Black Creek Canal virus, in experimentally infected Sigmodon hispidus. Am J Trop Med Hyg 59 : 58–65.
  10. Garin D, Peyrefitte C, Crance JM, Le Faou A, Jouan A, Bouloy M, 2001. Highly sensitive TaqMan PCR detection of Puumala hantavirus. Microbes Infect 3 : 739–745.
  11. Hutchinson KL, Peters CJ, Nichol ST, 1996. Sin Nombre virus mRNA synthesis. Virology 224 : 139–149.
  12. Thompson WH, Kalfayan B, Anslow RO, 1965. Isolation of California encephalitis group virus from fatal human illness. Am J Epidemiol 81 : 245–253.
  13. Karber G, 1931. Bietrag zur kollktiven behandlung pharmakologischer reiheversuche (On collective treatment of serial pharmacologic studies). Naunyn-Schmiedebergs Arch Exp Pathol Pharmakol 162 : 480–483.
  14. Kascsak RJ, Lyons MJ, 1978. Bunyamwera virus II. The generation and nature of defective interfering particles. Virology 89 : 539–546.
  15. Elliott RM, Wilkie ML, 1986. Persistent infection of Aedes albopictus C6/36 cells by bunyamwera virus. Virology 150 : 21–32.
  16. Chandler LJ, Beaty BJ, Baldridge GD, Bishop DHL, Hewlett MJ, 1990. Heterologous reassortment of bunyaviruses in Aedes triseriatus mosquitoes and transovarial and oral transmission of newly evolved genotypes. J Gen Virol 72 : 1045–1050.
  17. Dobie DK, Blair CD, Chandler LJ, Rayms-Keller A, McGaw MM, Wasieloski LP, Beaty BJ, 1997. Analysis of La crosse virus S mRNA 5′ termini in infected mosquito cells and Aedes triseriatus mosquitoes. J Virol 71 : 4395–4399.
  18. Chomczynski P, Sacchi N, 1987. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162 : 156–159.
  19. Scheffe H, 1953. A method for judging all contrasts in the analysis of variance. Biometrika 40 : 87–104.
  20. Tukey J, 1994. The Collected Works of John W. Tukey: Multiple Comparisons: 1948–1983. New York, Chapman and Hall.
  21. Kempf B, 2003. PhD Dissertation, Colorado State University.
  22. Beaty BJ, Thompson WH, 1978. Tropisms of La Crosse virus in Aedes triseriatus (Diptera: Culicidae) following infective blood meals. J Med Entomol 14 : 499–503.
  23. Chandler LJ, Blair CD, Beaty BJ, 1998. La Crosse virus infection of Aedes triseriatus (Diptera: Culicidae) ovaries before dissemination of virus from the midgut. J Med Entomol 35 : 567–572.
  24. Miller BR, DeFoliart GR, Yuill TM, 1979. Aedes triseriatus and La Crosse virus: lack of infection in eggs of the first ovarian cycle following oral infection of females. Am J Trop Med Hyg 28 : 897–901.
  25. Pantuwatana S, Thompson WH, Watts DM, Hanson RP, 1972. Experimental infection of chipmunks and squirrels with La Crosse and Trivittatus viruses and biological transmission of La Crosse virus by Aedes triseriatus. Am J Trop Med Hyg 21 : 476–481.
  26. Kirkpatrick BA, Washburn JO, Engelhard EK, Volkman LE, 1994. Primary infection of insect tracheae by Autographa californica M nuclear polyhedrosis virus. Virology 203 : 184–186.
  27. Myles KM, Pierro DJ, Olson KE, 2003. Deletions in the putative cell receptor-binding domain of Sindbis virus strain MRE16 E2 glycoprotein reduce midgut infectivity in Aedes aegypti. J Virol 77 : 8872–8881.
  28. Hardy J, 1988. Susceptibility and resistance of vector mosquitoes. Monath TP, ed. The Arboviruses: Epidemiology and Ecology. Vol. I. Boca Raton: CRC Press, 87–126.
  29. Woodring JL, Higgs S, Beaty BJ, 1996. Natural cycles of vector-borne pathogens. Beaty BJ, Marquardt WC, eds. The Biology of Disease Vectors. Niwot, CO: University Press of Colorado, 51–72.
  30. Weaver SC, Scott TW, 1990. Peritrophic membrane formation and cellular turnover in the midgut of Culiseta melanura (Diptera: Culicidae). J Med Entomol 27 : 864–873.
  31. Weaver SC, Scott TW, Lorenz LH, Repik PM, 1991. Detection of eastern equine encephalomyelitis virus deposition in Culiseta melanura following ingestion of radiolabeled virus in blood meals. Am J Trop Med Hyg 44 : 250–259.
  32. Anderson WA, Spielman A, 1971. Permeability of the ovarian follicle of Aedes aegypti mosquitoes. J Cell Biol 50 : 201–221.
  33. Clements AN, 1992. The Biology of Mosquitoes. Vol. I. Development, Nutrition, and Reproduction. New York: Chapman and Hall.
  34. Clements AN, Boocock MR, 1984. Ovarian development in mosquitoes: stages of growth and arrest, and follicular resorption. Physiol Entomol 9 : 1–8.
  35. Koller CN, Dhadialla TS, Raikhel A, 1989. Selective endocytosis of vitellogenin by oocytes of the mosquito, Aedes aegypti: an in vitro study. Insect Biochem 19 : 693–702.
  36. Banks GK, Malcolm CA, Clements AN, 1994. Temporal pattern of RNA and protein synthesis in the ovary of Aedes aegypti. J Insect Physiol 40 : 463–468.
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  • Received : 17 Feb 2004
  • Accepted : 05 Feb 2005

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