Skip to main content
NCBI home page
Applied Microbiology logoLink to Applied Microbiology
. 1970 Aug;20(2):233–239. doi: 10.1128/am.20.2.233-239.1970

Rapid Detection and Identification of Respiratory Viruses by Direct Immunofluorescence

Donn D'Alessio 1, Stanley Williams 1, Elliot C Dick 1
PMCID: PMC376907  PMID: 4098101

Abstract

The use of fluorescein-conjugated antiserum against respiratory syncytial (RS) and parainfluenza 1 and 3 viruses was compared with conventional techniques in the rapid detection of virus in tissue cultures inoculated with pharyngeal specimens known to contain these viruses. Twenty-three specimens were tested: 9 RS, 8 parainfluenza 1, and 6 parainfluenza 3. The fluorescent-antibody technique (FA) detected virus in 52% of the tissue cultures in 24 hr, and, by 72 hr, 22 of the 23 cultures were FA-positive whereas only 5 were positive by conventional techniques. Additionally, conjugated antisera were prepared against herpes simplex, influenza A2, and adenovirus type 5. All conjugates stained only the homologous virus and were 100- to 10,000-fold more sensitive than conventional techniques in detecting descending dilutions of virus inocula by 24 hr. With the procedures described, several antisera could be conjugated and ready for use within 24 hr. Serum fractionation was by ammonium sulfate precipitation, and with the procedure outlined virtually complete recovery of the globulin fraction and elimination of all of the albumin were accomplished.

Full text

PDF
233

Images in this article

236Image
on p.236
237Image
on p.237
237Image
on p.237

Selected References

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

  1. BAUMSTARK J. S., LAFFIN R. J., BARDAWIL W. A. A PREPARATIVE METHOD FOR THE SEPARATION OF 7S GAMMA GLOBULIN FROM HUMAN SERUM. Arch Biochem Biophys. 1964 Dec;108:514–522. doi: 10.1016/0003-9861(64)90436-9. [DOI] [PubMed] [Google Scholar]
  2. CHANOCK R. M., PARROTT R. H. ACUTE RESPIRATORY DISEASE IN INFANCY AND CHILDHOOD: PRESENT UNDERSTANDING AND PROSPECTS FOR PREVENTION. Pediatrics. 1965 Jul;36:21–39. [PubMed] [Google Scholar]
  3. CLARK H. F., SHEPARD C. C. A DIALYSIS TECHNIQUE FOR PREPARING FLUORESCENT ANTIBODY. Virology. 1963 Aug;20:642–644. doi: 10.1016/0042-6822(63)90292-7. [DOI] [PubMed] [Google Scholar]
  4. DEDMON R. E., HOLMES A. W., DEINHARDT F. PREPARATION OF FLUORESCEIN ISOTHIOCYANATE-LABELED GAMMA-GLOBULIN BY DIALYSIS, GEL FILTRATION, AND IONEXCHANGE CHROMATOGRAPHY IN COMBINATION. J Bacteriol. 1965 Mar;89:734–739. doi: 10.1128/jb.89.3.734-739.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DICK E. C., MOGABGAB W. J. Characteristics of para-influenza 1 (HA-2) virus. II. Hemagglutination, hemagglutination-inhibition and neutralization. Am J Hyg. 1961 May;73:273–281. doi: 10.1093/oxfordjournals.aje.a120186. [DOI] [PubMed] [Google Scholar]
  6. Dick E. C., Blumer C. R., Evans A. S. Epidemiology of infections with rhinovirus types 43 and 55 in a group of university of Wisconsin student families. Am J Epidemiol. 1967 Sep;86(2):386–400. doi: 10.1093/oxfordjournals.aje.a120749. [DOI] [PubMed] [Google Scholar]
  7. GOLDSTEIN G., SPALDING B. H., HUNT W. B., Jr Studies on fluorescent antibody staining II. Inhibition by sub- optimally conjugated antibody globulins. Proc Soc Exp Biol Med. 1962 Nov;111:416–421. doi: 10.3181/00379727-111-27810. [DOI] [PubMed] [Google Scholar]
  8. Gardner P. S., McQuilin J. Viral diagnosis by immunofluorescence. Lancet. 1968 Mar 16;1(7542):597–598. doi: 10.1016/s0140-6736(68)92876-6. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Hers J. F., van der Kuip L., Masurel N. Rapid diagnosis of influenza. Lancet. 1968 Mar 9;1(7541):510–511. doi: 10.1016/s0140-6736(68)91470-0. [DOI] [PubMed] [Google Scholar]
  11. Hsiung G. D. Latent virus infections in primate tissues with special reference to simian viruses. Bacteriol Rev. 1968 Sep;32(3):185–205. doi: 10.1128/br.32.3.185-205.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KAUFMAN L., CHERRY W. B. Technical factors affecting the preparation of fluorescent antibody reagents. J Immunol. 1961 Jul;87:72–79. [PubMed] [Google Scholar]
  13. LEVY H. B., SOBER H. A. A simple chromatographic method for preparation of gamma globulin. Proc Soc Exp Biol Med. 1960 Jan;103:250–252. doi: 10.3181/00379727-103-25476. [DOI] [PubMed] [Google Scholar]
  14. LEWIS V. J., JONES W. L., BROOKS J. B., CHERRY W. B. TECHNICAL CONSIDERATIONS IN THE PREPARATION OF FLUORESCENT-ANTIBODY CONJUGATES. Appl Microbiol. 1964 Jul;12:343–348. doi: 10.1128/am.12.4.343-348.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LIU C. Rapid diagnosis of human influenza infection from nasal smears by means of fluorescein-labeled antibody. Proc Soc Exp Biol Med. 1956 Aug-Sep;92(4):883–887. doi: 10.3181/00379727-92-22642. [DOI] [PubMed] [Google Scholar]
  16. MARSHALL J. D., EVELAND W. C., SMITH C. W. Superiority of fluorescein isothiocyanate (Riggs) for fluorescent-antibody technic with a modification of its application. Proc Soc Exp Biol Med. 1958 Aug-Sep;98(4):898–900. doi: 10.3181/00379727-98-24222. [DOI] [PubMed] [Google Scholar]
  17. McQuillin J., Gardner P. S. Rapid diagnosis of respiratory syncytial virus infection by immunofluorescent antibody techniques. Br Med J. 1968 Mar 9;1(5592):602–605. doi: 10.1136/bmj.1.5592.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SMITH C. W., MARSHALL J. D., Jr, EVELAND W. C. Use of contrasting fluorescent dye as counterstain in fixed tissue preparations. Proc Soc Exp Biol Med. 1959 Oct;102:179–181. doi: 10.3181/00379727-102-25182. [DOI] [PubMed] [Google Scholar]
  19. Schieble J. H., Lennette E. H., Kase A. An immunofluorescent staining method for rapid identification of respiratory syncytial virus. Proc Soc Exp Biol Med. 1965 Oct;120(1):203–208. doi: 10.3181/00379727-120-30486. [DOI] [PubMed] [Google Scholar]
  20. Spendlove R. S. Optimal labeling of antibody with fluorescein isothiocyanate. Proc Soc Exp Biol Med. 1966 Jun;122(2):580–583. doi: 10.3181/00379727-122-31196. [DOI] [PubMed] [Google Scholar]
  21. Wells A. F., Miller C. E., Nadel M. K. Rapid fluorescein and protein assay method for fluorescent-antibody conjugates. Appl Microbiol. 1966 Mar;14(2):271–275. doi: 10.1128/am.14.2.271-275.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wood B. T., Thompson S. H., Goldstein G. Fluorescent antibody staining. 3. Preparation of fluorescein-isothiocyanate-labeled antibodies. J Immunol. 1965 Aug;95(2):225–229. [PubMed] [Google Scholar]

Articles from Applied Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES