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. 1989 Nov;27(11):2505–2508. doi: 10.1128/jcm.27.11.2505-2508.1989

Detection of influenza virus by centrifugal inoculation of MDCK cells and staining with monoclonal antibodies.

R D Mills 1, K J Cain 1, G L Woods 1
PMCID: PMC267067  PMID: 2808674

Abstract

Two methods for detection of influenza virus in 451 clinical respiratory specimens were compared: (i) 24-well-plate centrifugation with Madin-Darby canine kidney (MDCK) cells and staining with monoclonal antibody pools to influenza viruses A and B (Centers for Disease Control, Atlanta, Ga.) in an indirect immunofluorescence assay after incubation for 40 h, and (ii) conventional tissue cell culture with primary monkey cells and hemadsorption. For 100 of these specimens, direct examination of smears by the direct fluorescence assay with monoclonal antibodies (Boots Cell Tech/API Analytab Products, Plainview, N.Y.) was also performed. Influenza A virus was recovered from 28 specimens by tissue cell culture after incubation for an average of 4.75 days (range, 2 to 14 days). Influenza B virus was recovered from 35 specimens by tissue culture after incubation for an average of 5.4 days (range, 3 to 14 days). By the centrifugation assay, 23 specimens were positive for influenza A virus and 30 were positive for influenza B virus. All specimens positive by the centrifugation assay were also positive by conventional tissue cell culture. The sensitivities of the centrifugation assay were 82% for detection of influenza A virus and 86% for influenza B virus (84% overall); the specificity of the assay was 100%. Of the 100 specimens studied by direct examination, 15 were positive for influenza virus by both conventional culture and centrifugation assay; however, the direct-smear results for these 15 specimens were negative in 13 cases and inconclusive in 2. The centrifugation assay is a rapid and specific method for detection of influenza A and B viruses in clinical specimens, and it can serve as a valuable and cost-efficient adjunct to conventional culture methods.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Daisy J. A., Lief F. S., Friedman H. M. Rapid diagnosis of influenza A infection by direct immunofluorescence of nasopharyngeal aspirates in adults. J Clin Microbiol. 1979 Jun;9(6):688–692. doi: 10.1128/jcm.9.6.688-692.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Espy M. J., Smith T. F., Harmon M. W., Kendal A. P. Rapid detection of influenza virus by shell vial assay with monoclonal antibodies. J Clin Microbiol. 1986 Oct;24(4):677–679. doi: 10.1128/jcm.24.4.677-679.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Frank A. L., Couch R. B., Griffis C. A., Baxter B. D. Comparison of different tissue cultures for isolation and quantitation of influenza and parainfluenza viruses. J Clin Microbiol. 1979 Jul;10(1):32–36. doi: 10.1128/jcm.10.1.32-36.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gleaves C. A., Smith T. F., Shuster E. A., Pearson G. R. Comparison of standard tube and shell vial cell culture techniques for the detection of cytomegalovirus in clinical specimens. J Clin Microbiol. 1985 Feb;21(2):217–221. doi: 10.1128/jcm.21.2.217-221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grandien M., Pettersson C. A., Gardner P. S., Linde A., Stanton A. Rapid viral diagnosis of acute respiratory infections: comparison of enzyme-linked immunosorbent assay and the immunofluorescence technique for detection of viral antigens in nasopharyngeal secretions. J Clin Microbiol. 1985 Nov;22(5):757–760. doi: 10.1128/jcm.22.5.757-760.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Harmon M. W., Pawlik K. M. Enzyme immunoassay for direct detection of influenza type A and adenovirus antigens in clinical specimens. J Clin Microbiol. 1982 Jan;15(1):5–11. doi: 10.1128/jcm.15.1.5-11.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Meguro H., Bryant J. D., Torrence A. E., Wright P. F. Canine kidney cell line for isolation of respiratory viruses. J Clin Microbiol. 1979 Feb;9(2):175–179. doi: 10.1128/jcm.9.2.175-179.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Minnich L. L., Ray C. G. Early testing of cell cultures for detection of hemadsorbing viruses. J Clin Microbiol. 1987 Feb;25(2):421–422. doi: 10.1128/jcm.25.2.421-422.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Paul W. S., Cowan J., Jackson G. G. Acute respiratory illness among immunized and nonimmunized patients with high-risk factors during a split season of influenza A and B. J Infect Dis. 1988 Apr;157(4):633–639. doi: 10.1093/infdis/157.4.633. [DOI] [PubMed] [Google Scholar]
  10. Shalit I., McKee P. A., Beauchamp H., Waner J. L. Comparison of polyclonal antiserum versus monoclonal antibodies for the rapid diagnosis of influenza A virus infections by immunofluorescence in clinical specimens. J Clin Microbiol. 1985 Nov;22(5):877–879. doi: 10.1128/jcm.22.5.877-879.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Stokes C. E., Bernstein J. M., Kyger S. A., Hayden F. G. Rapid diagnosis of influenza A and B by 24-h fluorescent focus assays. J Clin Microbiol. 1988 Jul;26(7):1263–1266. doi: 10.1128/jcm.26.7.1263-1266.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Swenson P. D., Kaplan M. H. Rapid detection of influenza virus in cell culture by indirect immunoperoxidase staining with type-specific monoclonal antibodies. Diagn Microbiol Infect Dis. 1987 Aug;7(4):265–268. doi: 10.1016/0732-8893(87)90142-8. [DOI] [PubMed] [Google Scholar]
  13. Thiele G. M., Bicak M. S., Young A., Kinsey J., White R. J., Purtilo D. T. Rapid detection of cytomegalovirus by tissue culture, centrifugation, and immunofluorescence with a monoclonal antibody to an early nuclear antigen. J Virol Methods. 1987 Jul;16(4):327–338. doi: 10.1016/0166-0934(87)90018-8. [DOI] [PubMed] [Google Scholar]
  14. Walls H. H., Harmon M. W., Slagle J. J., Stocksdale C., Kendal A. P. Characterization and evaluation of monoclonal antibodies developed for typing influenza A and influenza B viruses. J Clin Microbiol. 1986 Feb;23(2):240–245. doi: 10.1128/jcm.23.2.240-245.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Woods G. L., Johnson A. M. Rapid 24-well plate centrifugation assay for detection of influenza A virus in clinical specimens. J Virol Methods. 1989 Apr-May;24(1-2):35–42. doi: 10.1016/0166-0934(89)90005-0. [DOI] [PubMed] [Google Scholar]
  16. Woods G. L., Young A., Johnson A., Thiele G. M. Detection of cytomegalovirus by 24-well plate centrifugation assay using a monoclonal antibody to an early nuclear antigen and by conventional cell culture. J Virol Methods. 1987 Dec;18(4):207–213. doi: 10.1016/0166-0934(87)90082-6. [DOI] [PubMed] [Google Scholar]

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