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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1990 Sep;28(9):1952–1956. doi: 10.1128/jcm.28.9.1952-1956.1990

Detection of bluetongue virus serotype 17 in Culicoides variipennis by nucleic acid blot and sandwich hybridization techniques.

R J Schoepp 1, J F Bray 1, K E Olson 1, A el-Hussein 1, F R Holbrook 1, C D Blair 1, P Roy 1, B J Beaty 1
PMCID: PMC268085  PMID: 2172294

Abstract

Molecular hybridization techniques were developed for the detection and surveillance of bluetongue virus (BTV) serotype 17 in the insect vector Culicoides variipennis, a biting midge. Radiolabeled RNA and cDNA probes were generated from sequences of the L3 segment of BTV serotype 17. These probes were used to detect BTV RNA in pools of infected C. variipennis by hybridizing the probes directly to analyte immobilized on nylon membranes or by using a nucleic acid sandwich hybridization test. Hybridization procedures were able to detect 1 infected C. variipennis in a pool of 50 and as little as 3.55 log10 50% tissue culture infective doses per ml of virus. These hybridization techniques provide an alternative to virus isolation for the surveillance of BTV in vector populations.

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Selected References

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  1. Barber T. L. Temporal appearance, geographic distribution, and species of origin of bluetongue virus serotypes in the United States. Am J Vet Res. 1979 Nov;40(11):1654–1656. [PubMed] [Google Scholar]
  2. Bowne J. G. Bluetongue disease. Adv Vet Sci Comp Med. 1971;15:1–46. [PubMed] [Google Scholar]
  3. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dangler C. A., Dunn S. J., Squire K. R., Stott J. L., Osburn B. I. Rapid identification of bluetongue virus by nucleic acid hybridization in solution. J Virol Methods. 1988 Aug;20(4):353–365. doi: 10.1016/0166-0934(88)90138-3. [DOI] [PubMed] [Google Scholar]
  5. Gibbs E. P., Greiner E. C., Taylor W. P., Barber T. L., House J. A., Pearson J. E. Isolation of bluetongue virus serotype 2 from cattle in Florida: serotype of bluetongue virus hitherto unrecognized in the Western Hemisphere. Am J Vet Res. 1983 Dec;44(12):2226–2228. [PubMed] [Google Scholar]
  6. Howell P. G., Verwoerd D. W. Bluetongue virus. Virol Monogr. 1971;9:35–74. doi: 10.1007/978-3-7091-3987-5_2. [DOI] [PubMed] [Google Scholar]
  7. Jochim M. M., Jones R. H. Multiplication of bluetongue virus in Culicoides variipennis following artificial infection. Am J Epidemiol. 1966 Sep;84(2):241–246. doi: 10.1093/oxfordjournals.aje.a120637. [DOI] [PubMed] [Google Scholar]
  8. Jones R. H., Foster N. M. Relevance of laboratory colonies of the vector in arbovirus research--Culicoides variipennis and bluetongue. Am J Trop Med Hyg. 1978 Jan;27(1 Pt 1):168–177. doi: 10.4269/ajtmh.1978.27.168. [DOI] [PubMed] [Google Scholar]
  9. Lehtomäki K., Julkunen I., Sandelin K., Salonen J., Virtanen M., Ranki M., Hovi T. Rapid diagnosis of respiratory adenovirus infections in young adult men. J Clin Microbiol. 1986 Jul;24(1):108–111. doi: 10.1128/jcm.24.1.108-111.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lomeli H., Tyagi S., Pritchard C. G., Lizardi P. M., Kramer F. R. Quantitative assays based on the use of replicatable hybridization probes. Clin Chem. 1989 Sep;35(9):1826–1831. [PubMed] [Google Scholar]
  11. Luedke A. J., Jones R. H. Storage of bluetongue virus-infected Culicoides variipennis. Am J Vet Res. 1972 Sep;33(9):1875–1878. [PubMed] [Google Scholar]
  12. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Olson K., Blair C., Padmanabhan R., Beaty B. Detection of dengue virus type 2 in Aedes albopictus by nucleic acid hybridization with strand-specific RNA probes. J Clin Microbiol. 1988 Mar;26(3):579–581. doi: 10.1128/jcm.26.3.579-581.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Purdy M., Petre J., Roy P. Cloning of the bluetongue virus L3 gene. J Virol. 1984 Sep;51(3):754–759. doi: 10.1128/jvi.51.3.754-759.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ranki M., Palva A., Virtanen M., Laaksonen M., Söderlund H. Sandwich hybridization as a convenient method for the detection of nucleic acids in crude samples. Gene. 1983 Jan-Feb;21(1-2):77–85. doi: 10.1016/0378-1119(83)90149-x. [DOI] [PubMed] [Google Scholar]
  16. Rotbart H. A., Levin M. J., Murphy N. L., Abzug M. J. RNA target loss during solid phase hybridization of body fluids--a quantitative study. Mol Cell Probes. 1987 Dec;1(4):347–358. doi: 10.1016/0890-8508(87)90016-8. [DOI] [PubMed] [Google Scholar]
  17. Roy P. Bluetongue virus genetics and genome structure. Virus Res. 1989 Jul;13(3):179–206. doi: 10.1016/0168-1702(89)90015-4. [DOI] [PubMed] [Google Scholar]
  18. Roy P., Ritter G. D., Jr, Akashi H., Collisson E., Inaba Y. A genetic probe for identifying bluetongue virus infections in vivo and in vitro. J Gen Virol. 1985 Jul;66(Pt 7):1613–1619. doi: 10.1099/0022-1317-66-7-1613. [DOI] [PubMed] [Google Scholar]
  19. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  20. Virtanen M., Palva A., Laaksonen M., Halonen P., Söderlund H., Ranki M. Novel test for rapid viral diagnosis: detection of adenovirus in nasopharyngeal mucus aspirates by means of nucleic-acid sandwich hybridisation. Lancet. 1983 Feb 19;1(8321):381–383. doi: 10.1016/s0140-6736(83)91500-3. [DOI] [PubMed] [Google Scholar]
  21. Virtanen M., Syvänen A. C., Oram J., Söderlund H., Ranki M. Cytomegalovirus in urine: detection of viral DNA by sandwich hybridization. J Clin Microbiol. 1984 Dec;20(6):1083–1088. doi: 10.1128/jcm.20.6.1083-1088.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wang L., Kemp M. C., Roy P., Collisson E. W. Tissue tropism and target cells of bluetongue virus in the chicken embryo. J Virol. 1988 Mar;62(3):887–893. doi: 10.1128/jvi.62.3.887-893.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yolken R. H. Nucleic acids or immunoglobulins: which are the molecular probes of the future? Mol Cell Probes. 1988 Jun;2(2):87–96. doi: 10.1016/0890-8508(88)90030-8. [DOI] [PubMed] [Google Scholar]
  24. de Mattos C. A., de Mattos C. C., Osburn B. I. Recombinant cDNA probe from bluetongue virus genome segment 10 for identification of bluetongue virus. J Vet Diagn Invest. 1989 Jul;1(3):237–241. doi: 10.1177/104063878900100308. [DOI] [PubMed] [Google Scholar]
  25. de Mattos C. C., de Mattos C. A., Osburn B. I., Dangler C. A., Chuang R. Y., Doi R. H. Recombinant DNA probe for serotype-specific identification of bluetongue virus 17. Am J Vet Res. 1989 Apr;50(4):536–541. [PubMed] [Google Scholar]

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