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. 1988 Mar;26(3):588–591. doi: 10.1128/jcm.26.3.588-591.1988

Detection of respiratory syncytial virus in clinical specimens by viral culture, direct and indirect immunofluorescence, and enzyme immunoassay.

J H Hughes 1, D R Mann 1, V V Hamparian 1
PMCID: PMC266341  PMID: 3281981

Abstract

We evaluated prospectively the detection of respiratory syncytial virus (RSV) by culture and by direct antigen detection using an indirect immunofluorescence assay (IFA), a direct monoclonal immunofluorescence assay (DFA), and a monoclonal enzyme immunoassay (EIA). Of 221 specimens, 95 (43%) were culture positive for RSV, 4 (1.8%) contained more than one virus, and 17 (7.6%) contained a virus other than RSV. Overall, HEp-2 and Flow 6000 cells grew significantly more RSV isolates (82 and 72%, respectively) than A549 cells, which grew only 29% of the isolates. The mean time for RSV detection with HEp-2 cells was 2.9 days. This was significantly less than the mean time for RSV detection with either Flow 6000 cells (6.1 days) or A549 cells (6.4 days). Of 221 specimens, 129 were tested simultaneously by culture, IFA, and DFA. Of these 129 specimens, 62 (48%) were positive by culture, 69 (53%) were positive by IFA, and 70 (54%) were positive by DFA. For 92 specimens screened simultaneously by culture, IFA, and EIA, positive results were obtained for 33 (36%) of the specimens by both culture and IFA and for 29 (32%) of the specimens by EIA. Of 126 culture-negative specimens, 21 (17%) were positive for RSV when determined by IFA. Conversely, 14 (15%) of 95 RSV culture-positive specimens were negative by IFA, whereas DFA missed 19% of the culture-positive specimens. Compared with culture, the Kallestad EIA kit had a sensitivity and specificity of 73 and 92% respectively, but missed 9 (27%) of 33 culture-positive specimens. These data demonstrate that isolation by culture continues to be important for viral diagnosis of REV infections and that for valid comparative studies between viral isolations and rapid detection methods, both sensitive host cells and appropriate test conditions are required.

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Selected References

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  1. Ahluwalia G., Embree J., McNicol P., Law B., Hammond G. W. Comparison of nasopharyngeal aspirate and nasopharyngeal swab specimens for respiratory syncytial virus diagnosis by cell culture, indirect immunofluorescence assay, and enzyme-linked immunosorbent assay. J Clin Microbiol. 1987 May;25(5):763–767. doi: 10.1128/jcm.25.5.763-767.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anestad G., Breivik N., Thoresen T. Rapid diagnosis of respiratory syncytial virus and influenza A virus infections by immunofluorescence: experience with a simplified procedure for the preparation of cell smears from nasopharyngeal secretions. Acta Pathol Microbiol Immunol Scand B. 1983 Aug;91(4):267–271. doi: 10.1111/j.1699-0463.1983.tb00045.x. [DOI] [PubMed] [Google Scholar]
  3. Anestad G. Surveillance of respiratory viral infections by rapid immunofluorescence diagnosis, with emphasis on the epidemiological development of respiratory syncytial virus infections. J Hyg (Lond) 1985 Jun;94(3):349–356. doi: 10.1017/s0022172400061581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arens M. Q., Swierkosz E. M., Schmidt R. R., Armstrong T., Rivetna K. A. Enhanced isolation of respiratory syncytial virus in cell culture. J Clin Microbiol. 1986 Apr;23(4):800–802. doi: 10.1128/jcm.23.4.800-802.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Battaglia M. F., Balducci L. Rapid diagnosis of respiratory syncytial virus infections by immunofluorescence on cell smears from nasopharyngeal secretions. Boll Ist Sieroter Milan. 1980 May 31;59(2):121–125. [PubMed] [Google Scholar]
  6. Bell D. M., Walsh E. E., Hruska J. F., Schnabel K. C., Hall C. B. Rapid detection of respiratory syncytial virus with a monoclonal antibody. J Clin Microbiol. 1983 Jun;17(6):1099–1101. doi: 10.1128/jcm.17.6.1099-1101.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bromberg K., Tannis G., Daidone B., Clarke L., Sierra M. F. Comparison of ortho respiratory syncytial virus enzyme-linked immunosorbent assay and HEp-2 cell culture. J Clin Microbiol. 1985 Dec;22(6):1071–1072. doi: 10.1128/jcm.22.6.1071-1072.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bromberg K., Tannis G., Daidone B., Clarke L., Sierra M. Comparison of HEp-2 cell culture and Abbott respiratory syncytial virus enzyme immunoassay. J Clin Microbiol. 1987 Feb;25(2):434–436. doi: 10.1128/jcm.25.2.434-436.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cevenini R., Donati M., Moroni A., Franchi L., Rumpianesi F. Rapid immunoperoxidase assay for detection of respiratory syncytial virus in nasopharyngeal secretions. J Clin Microbiol. 1983 Oct;18(4):947–949. doi: 10.1128/jcm.18.4.947-949.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chao R. K., Fishaut M., Schwartzman J. D., McIntosh K. Detection of respiratory syncytial virus in nasal secretions from infants by enzyme-linked immunosorbent assay. J Infect Dis. 1979 Apr;139(4):483–486. doi: 10.1093/infdis/139.4.483. [DOI] [PubMed] [Google Scholar]
  11. Cheeseman S. H., Pierik L. T., Leombruno D., Spinos K. E., McIntosh K. Evaluation of a commercially available direct immunofluorescent staining reagent for the detection of respiratory syncytial virus in respiratory secretions. J Clin Microbiol. 1986 Jul;24(1):155–156. doi: 10.1128/jcm.24.1.155-156.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chonmaitree T., Bessette-Henderson B. J., Hepler R. E., Lucia H. L. Comparison of three rapid diagnostic techniques for detection of respiratory syncytial virus from nasal wash specimens. J Clin Microbiol. 1987 Apr;25(4):746–747. doi: 10.1128/jcm.25.4.746-747.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cradock-Watson J. E., McQuillin J., Gardner P. S. Rapid diagnosis of respiratory syncytial virus infection in children by the immunofluorescent technique. J Clin Pathol. 1971 May;24(4):308–312. doi: 10.1136/jcp.24.4.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Flanders R. T., Lindsay P. D., Chairez R., Brawner T. A., Kumar M. L., Swenson P. D., Bromberg K. Evaluation of clinical specimens for the presence of respiratory syncytial virus antigens using an enzyme immunoassay. J Med Virol. 1986 May;19(1):1–9. doi: 10.1002/jmv.1890190102. [DOI] [PubMed] [Google Scholar]
  15. Freke A., Stott E. J., Roome A. P., Caul E. O. The detection of respiratory syncytial virus in nasopharyngeal aspirates: assessment, formulation, and evaluation of monoclonal antibodies as a diagnostic reagent. J Med Virol. 1986 Feb;18(2):181–191. doi: 10.1002/jmv.1890180210. [DOI] [PubMed] [Google Scholar]
  16. Freymuth F., Quibriac M., Petitjean J., Amiel M. L., Pothier P., Denis A., Duhamel J. F. Comparison of two new tests for rapid diagnosis of respiratory syncytial virus infections by enzyme-linked immunosorbent assay and immunofluorescence techniques. J Clin Microbiol. 1986 Dec;24(6):1013–1016. doi: 10.1128/jcm.24.6.1013-1016.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gardner P. S., McQuillin J. Application of immunofluorescent antibody technique in rapid diagnosis of respiratory syncytial virus infection. Br Med J. 1968 Aug 10;3(5614):340–343. doi: 10.1136/bmj.3.5614.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Gray K. G., MacFarlane D. E., Sommerville R. G. Direct immunofluorescent identification of respiratory syncytial virus in throat swabs from children with respiratory illness. Lancet. 1968 Mar 2;1(7540):446–448. doi: 10.1016/s0140-6736(68)92779-7. [DOI] [PubMed] [Google Scholar]
  20. Hall C. B., Douglas R. G., Jr Clinically useful method for the isolation of respiratory syncytial virus. J Infect Dis. 1975 Jan;131(1):1–5. doi: 10.1093/infdis/131.1.1. [DOI] [PubMed] [Google Scholar]
  21. Hall C. B., Douglas R. G., Jr, Geiman J. M., Messner M. K. Nosocomial respiratory syncytial virus infections. N Engl J Med. 1975 Dec 25;293(26):1343–1346. doi: 10.1056/NEJM197512252932604. [DOI] [PubMed] [Google Scholar]
  22. Hall C. B., Douglas R. G., Jr, Geiman J. M. Respiratory syncytial virus infections in infants: quantitation and duration of shedding. J Pediatr. 1976 Jul;89(1):11–15. doi: 10.1016/s0022-3476(76)80918-3. [DOI] [PubMed] [Google Scholar]
  23. Hall C. B., Kopelman A. E., Douglas R. G., Jr, Geiman J. M., Meagher M. P. Neonatal respiratory syncytial virus infection. N Engl J Med. 1979 Feb 22;300(8):393–396. doi: 10.1056/NEJM197902223000803. [DOI] [PubMed] [Google Scholar]
  24. Hall C. B., McBride J. T., Gala C. L., Hildreth S. W., Schnabel K. C. Ribavirin treatment of respiratory syncytial viral infection in infants with underlying cardiopulmonary disease. JAMA. 1985 Dec 6;254(21):3047–3051. [PubMed] [Google Scholar]
  25. Hall C. B., McBride J. T., Walsh E. E., Bell D. M., Gala C. L., Hildreth S., Ten Eyck L. G., Hall W. J. Aerosolized ribavirin treatment of infants with respiratory syncytial viral infection. A randomized double-blind study. N Engl J Med. 1983 Jun 16;308(24):1443–1447. doi: 10.1056/NEJM198306163082403. [DOI] [PubMed] [Google Scholar]
  26. Hendry R. M., Fernie B. F., Anderson L. J., Godfrey E., McIntosh K. Monoclonal capture antibody ELISA for respiratory syncytial virus: detection of individual viral antigens and determination of monoclonal antibody specificities. J Immunol Methods. 1985 Mar 18;77(2):247–258. doi: 10.1016/0022-1759(85)90037-7. [DOI] [PubMed] [Google Scholar]
  27. Hendry R. M., McIntosh K. Enzyme-linked immunosorbent assay for detection of respiratory syncytial virus infection: development and description. J Clin Microbiol. 1982 Aug;16(2):324–328. doi: 10.1128/jcm.16.2.324-328.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Hendry R. M., Pierik L. T., McIntosh K. Comparison of washed nasopharyngeal cells and whole nasal secretions for detection of respiratory syncytial virus antigens by enzyme-linked immunosorbent assay. J Clin Microbiol. 1986 Feb;23(2):383–384. doi: 10.1128/jcm.23.2.383-384.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hornsleth A., Friis B., Andersen P., Brenøe E. Detection of respiratory syncytial virus in nasopharyngeal secretions by ELISA: comparison with fluorescent antibody technique. J Med Virol. 1982;10(4):273–281. doi: 10.1002/jmv.1890100407. [DOI] [PubMed] [Google Scholar]
  30. Hornsleth A., Friis B., Krasilnikof P. A. Detection of respiratory syncytial virus in nasopharyngeal secretions by a biotin-avidin ELISA more sensitive than the fluorescent antibody technique. J Med Virol. 1986 Feb;18(2):113–117. doi: 10.1002/jmv.1890180203. [DOI] [PubMed] [Google Scholar]
  31. Jalowayski A. A., England B. L., Temm C. J., Nunemacher T. J., Bastian J. F., MacPherson G. A., Dankner W. M., Straube R. C., Connor J. D. Peroxidase-antiperoxidase assay for rapid detection of respiratory syncytial virus in nasal epithelial specimens from infants and children. J Clin Microbiol. 1987 Apr;25(4):722–725. doi: 10.1128/jcm.25.4.722-725.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kao C. L., McIntosh K., Fernie B., Talis A., Pierik L., Anderson L. Monoclonal antibodies for the rapid diagnosis of respiratory syncytial virus infection by immunofluorescence. Diagn Microbiol Infect Dis. 1984 Jun;2(3):199–206. doi: 10.1016/0732-8893(84)90031-2. [DOI] [PubMed] [Google Scholar]
  33. Kaul A., Scott R., Gallagher M., Scott M., Clement J., Ogra P. L. Respiratory syncytial virus infection. Rapid diagnosis in children by use of indirect immunofluorescence. Am J Dis Child. 1978 Nov;132(11):1088–1090. doi: 10.1001/archpedi.1978.02120360044006. [DOI] [PubMed] [Google Scholar]
  34. Kim H. W., Wyatt R. G., Fernie B. F., Brandt C. D., Arrobio J. O., Jeffries B. C., Parrott R. H. Respiratory syncytial virus detection by immunofluorescence in nasal secretions with monoclonal antibodies against selected surface and internal proteins. J Clin Microbiol. 1983 Dec;18(6):1399–1404. doi: 10.1128/jcm.18.6.1399-1404.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Kumar M. L., Super D. M., Lembo R. M., Thomas F. C., Prokay S. L. Diagnostic efficacy of two rapid tests for detection of respiratory syncytial virus antigen. J Clin Microbiol. 1987 May;25(5):873–875. doi: 10.1128/jcm.25.5.873-875.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lauer B. A. Comparison of virus culturing and immunofluorescence for rapid detection of respiratory syncytial virus in nasopharyngeal secretions: sensitivity and specificity. J Clin Microbiol. 1982 Aug;16(2):411–412. doi: 10.1128/jcm.16.2.411-412.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lauer B. A., Masters H. A., Wren C. G., Levin M. J. Rapid detection of respiratory syncytial virus in nasopharyngeal secretions by enzyme-linked immunosorbent assay. J Clin Microbiol. 1985 Nov;22(5):782–785. doi: 10.1128/jcm.22.5.782-785.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Martin A. J., Gardner P. S., McQuillin J. Epidemiology of respiratory viral infection among paediatric inpatients over a six-year period in north-east England. Lancet. 1978 Nov 11;2(8098):1035–1038. doi: 10.1016/s0140-6736(78)92351-6. [DOI] [PubMed] [Google Scholar]
  39. McIntosh K., Hendry R. M., Fahnestock M. L., Pierik L. T. Enzyme-linked immunosorbent assay for detection of respiratory syncytial virus infection: application to clinical samples. J Clin Microbiol. 1982 Aug;16(2):329–333. doi: 10.1128/jcm.16.2.329-333.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Minnich L. L., Ray C. G. Comparison of direct and indirect immunofluorescence staining of clinical specimens for detection of respiratory syncytial virus antigen. J Clin Microbiol. 1982 May;15(5):969–970. doi: 10.1128/jcm.15.5.969-970.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Minnich L., Ray C. G. Comparison of direct immunofluorescent staining of clinical specimens for respiratory virus antigens with conventional isolation techniques. J Clin Microbiol. 1980 Sep;12(3):391–394. doi: 10.1128/jcm.12.3.391-394.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Mintz L., Ballard R. A., Sniderman S. H., Roth R. S., Drew W. L. Nosocomial respiratory syncytial virus infections in an intensive care nursery: rapid diagnosis by direct immunofluorescence. Pediatrics. 1979 Aug;64(2):149–153. [PubMed] [Google Scholar]
  43. Nagahama H., Eller J. J., Fulginiti V. A., Marks M. I. Direct immunofluorescent studies of infection with respiratory syncytial virus. J Infect Dis. 1970 Oct;122(4):260–271. doi: 10.1093/infdis/122.4.260. [DOI] [PubMed] [Google Scholar]
  44. Popow-Kraupp T., Kern G., Binder C., Tuma W., Kundi M., Kunz C. Detection of respiratory syncytial virus in nasopharyngeal secretions by enzyme-linked immunosorbent assay, indirect immunofluorescence, and virus isolation: a comparative study. J Med Virol. 1986 Jun;19(2):123–134. doi: 10.1002/jmv.1890190204. [DOI] [PubMed] [Google Scholar]
  45. Pothier P., Nicolas J. C., de Saint Maur G. P., Ghim S., Kazmierczak A., Bricout F. Monoclonal antibodies against respiratory syncytial virus and their use for rapid detection of virus in nasopharyngeal secretions. J Clin Microbiol. 1985 Feb;21(2):286–287. doi: 10.1128/jcm.21.2.286-287.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Ray C. G., Minnich L. L. Efficiency of immunofluorescence for rapid detection of common respiratory viruses. J Clin Microbiol. 1987 Feb;25(2):355–357. doi: 10.1128/jcm.25.2.355-357.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Routledge E. G., McQuillin J., Samson A. C., Toms G. L. The development of monoclonal antibodies to respiratory syncytial virus and their use in diagnosis by indirect immunofluorescence. J Med Virol. 1985 Mar;15(3):305–320. doi: 10.1002/jmv.1890150311. [DOI] [PubMed] [Google Scholar]
  48. Sarkkinen H. K., Halonen P. E., Arstila P. P., Salmi A. A. Detection of respiratory syncytial, parainfluenza type 2, and adenovirus antigens by radioimmunoassay and enzyme immunoassay on nasopharyngeal specimens from children with acute respiratory disease. J Clin Microbiol. 1981 Feb;13(2):258–265. doi: 10.1128/jcm.13.2.258-265.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Swenson P. D., Kaplan M. H. Rapid detection of respiratory syncytial virus in nasopharyngeal aspirates by a commercial enzyme immunoassay. J Clin Microbiol. 1986 Mar;23(3):485–488. doi: 10.1128/jcm.23.3.485-488.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Urquhart G. E., Walker G. H. Immunofluorescence for routine diagnosis of respiratory syncytial virus infection. J Clin Pathol. 1972 Oct;25(10):843–845. doi: 10.1136/jcp.25.10.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Vesikari T., Kuusela A. L., Sarkkinen H. K., Halonen P. E. Clinical evaluation of radioimmunoassay of nasopharyngeal secretions and serology for diagnosis of viral infections in children hospitalized for respiratory infections. Pediatr Infect Dis. 1982 Nov-Dec;1(6):391–394. doi: 10.1097/00006454-198211000-00006. [DOI] [PubMed] [Google Scholar]
  52. Waner J. L., Whitehurst N. J., Jonas S., Wall L., Shalaby H. Isolation of viruses from specimens submitted for direct immunofluorescence test for respiratory syncytial virus. J Pediatr. 1986 Feb;108(2):249–250. doi: 10.1016/s0022-3476(86)80993-3. [DOI] [PubMed] [Google Scholar]
  53. Wu B. The production of monoclonal antibodies against respiratory syncytial virus and its clinical applications. Clin Lab Med. 1985 Sep;5(3):589–613. [PubMed] [Google Scholar]

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