Abstract
In recent years immunoelectron microscopy (IEM) has become one of the several immunoassays available to virologists for detection and identification of viruses. Like other immunological assays, the reliability depends on careful attention to test conditions (e.g. antigen/antibody concentration, incubation time, temperature, pH) and to a regard for incorporation of a range of appropriate controls. This chapter will discuss several IEM techniques that can be applied to fluid specimens commonly encountered in diagnostic virology. Consideration will be given to some of the variables that affect the efficiency of these tests, and to the advantages and disadvantages of IEM relative to other immunoassays. Additional practical details are available from several published sources (Doane, 1974, 1986, 1987, 1988a, 1988b; Doane and Anderson, 1977, 1987; Katz and Kohn, 1984; Kjeldsberg, 1986).
Chapter PDF
Similar content being viewed by others
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
References
Almeida J, Cinader B, Howatson A (1963) The structure of antigen-antibody complexes. A study by electron microscopy. J Exp Med 118:327–340.
Almeida JD, Waterson AP (1969) The morphology of virus-antibody interaction. Adv Virus Res 15:307–338.
Anderson N, Doane FW (1972) Agar diffusion method for negative staining of microbial suspensions in salt solutions. Appl Microbiol 24:495–496.
Anderson N, Doane FW (1973) Specific identification of enteroviruses by immuno-electron microscopy using a serum-in-agar method. Can J Microbiol 19:585–589.
Anderson TF, Stanley WM (1941) A study by means of the electron microscope of the reaction between tobacco mosaic virus and its antiserum. J Biol Chem 139:339–344.
Bayer ME, Mannweiler E (1963) Antigen-antibody reactions in influenza virus as seen in the electron microscope. Arch Gesamte Virusforsch 13:541–547.
Bayer ME, Blumberg BS, Werner B (1968) Particles associated with Australia antigen in the sera of patients with leukemia, Down’s syndrome and hepatitis. Nature 218:1057–1059.
Berthiaume L, Alain R, McLaughlin B, Payment P (1981) Rapid detection of human viruses in faeces by a simple and routine immune electron microscopy technique. J Gen Virol 55:223–227.
Best JM, Bantvala JE, Almeida JD, Waterson AP (1967) Morphological characteristics of rubella virus. Lancet 2:237–239.
Brenner S, Horne RW (1959) A negative staining method for high resolution microscopy of viruses. Biochem Biophys Acta 34:103–110.
Derrick KS (1973) Quantitative assay for plant viruses using serological specific electron microscopy. Virology 56:652–653.
Doane FW (1974) Identification of viruses by immunoelectron microscopy. In: Kurstak E, Morisset R (eds) Viral Immunodiagnosis. Academic Press, New York, pp 237–255.
Doane FW (1986) Electron microscopy and immunoelectron microscopy. In: Specter S, Lancz GJ (eds) Clinical Virology Manual. Elsevier, New York, pp 71–88.
Doane FW (1987) Immunoelectron microscopy in diagnostic virology. Ultrastruct Pathol 11:681–685.
Doane FW (1988a) Immunoelectron microscopy and its role in diagnostic virology. Clin Immunol News 9:159–162.
Doane FW (1988b) Electron Microscopy. In: Lennette EH, Halonen P, Murphy FA (eds) Laboratory Diagnosis of Infectious Diseases. Principles and Practice. Springer-Verlag, New York, pp 121–131.
Doane FW, Anderson N (1977) Electron and immunoelectron microscopic procedures for diagnosis of viral infections. In Kurstak E, Kurstak C (eds) Comparative Diagnosis of Viral Diseases, Vol II, part B. Academic Press, New York, pp 505–539.
Doane FW, Anderson N (1987) Electron Microscopy in Diagnostic Virology: A Practical Guide and Atlas. Cambridge University Press, New York.
El-Ghorr AA, Snodgrass DR, Scott FMM (1988) Evaluation of an immunogold electron microscopy technique for detecting bovine coronavirus. J Virol Methods 19:215–224.
Faulk WP, Taylor GM (1971) An immunocolloidal method for the electron microscope. Immunochemistry 8:1081–1083.
Furui S (1986) Use of protein A in the serum-in-agar diffusion method in immune electron microscopy for detection of virus particles in cell culture. Microbiol Immunol 30:1023–1035
Gerna G, Passarani N, Battaglia M, Percivalle E (1984) Rapid serotyping of human rotavirus strains by solid-phase immune electron microscopy. J Clin Microbiol 19:273–278.
Gerna G, Passarani N, Sarasini A, Battaglia M (1985) Characterization of serotypes of human rotavirus strains by solid-phase immune electron microscopy. J Infect Dis 152:1143–1151.
Gerna G, Sarasini A, Coulson BS, Parea M, Torsellini M, Arbustini E, Battaglia M (1988) Comparative sensitivities of solid-phase immune electron microscopy and enzyme-linked immunosorbent assay for serotyping of human rotavirus strains with neutralizing monoclonal antibodies. J Clin Microbiol 26:1383–1387.
Hammond GW, Hazelton PR, Chuang I, Klisko, B. (1981) Improved detection of viruses by electron microscopy after direct ultracentrifuge preparation of specimens. J Clin Microbiol 14:220–221.
Hopley, JFA (1985) Protein A-gold immunoelectron microscopy studies on the simian rotavirus SAH. M.Sc thesis, University of Toronto.
Hopley JFA, Doane FW (1985) Development of a sensitive protein A-gold immunoelectron microscopy method for detecting viral antigens in fluid specimens. J Virol Methods 12:135–147.
Horisberger M (1981) Colloidal gold. A cytochemical marker for light and fluorescent microscopy and for transmission and scanning electronmi-croscopy. In: Johari O (ed) Scanning Electron Microscopy. AMF O’Hara, S.E.M., Inc., Chicago, vol II, pp 9–31.
Hummeler K, Anderson TF, Brown RA (1961) Identification of poliovirus particles of different antigenicity by specific agglutination as seen in the electron microscope. Virology 16:84–90.
Kapikian A, Almeida J, Stott E (1972a) Immune electron microscopy of rhino-viruses. J Virol 10:142–146.
Kapikian AZ, Wyatt RG, Dolin R, Thornhill TS, Kalica AR, Chanock RM (1972b) Visualization by immune electron microscopy of a 27 nm particle associated with acute infectious non-bacterial gastroenteritis. J Virol 10:1075–1081.
Kapikian A, James H, Kelly S, Vaughn A (1973) Detection of coronavirus 692 by immune electron microscopy. Infect Immun 7:111–116.
Kapikian AZ, Dienstag JL, Purcell RH (1976) Immune electron microscopy as a method for the detection, identification, and characterization of agents not cultivable in an in vitro system. In: Rose NR, Friedman H (eds) Manual of Clinical Immunology. Amer Soc Microbiol, Washington D.C. pp 467–480.
Katz D, Straussman Y, Shahar A, Kohn (1980) Solid-phase immune electron microscopy (SPIEM) for rapid viral diagnosis. J Immunol Methods 38:171–174.
Katz D, Kohn A (1984) Immunosorbent electron microscopy for detection of viruses. Adv Virus Res 29:169–194
Katz D, Straussman Y (1984) Evaluation of immunoadsorbent electron microscopic techniques for detection of Sindbis virus. J Virol Methods 8:243–254.
Kelen AE, Hathaway AE, McLeod DA (1971) Rapid detection of Australia/SH antigen and antibody by a simple and sensitive technique of immunoelectron microscopy. Can J Microbiol 17:993–1000.
Kjeldsberg E (1985) Specific labelling of human rotaviruses and adenoviruses with gold-IgG complexes. J Virol Methods 12:47–57.
Kjeldsberg E (1986) Immunonegative stain techniques for electron microscopic detection of viruses in human faeces. Ultrastr Pathol 10:553–570.
Kjeldsberg E, Mortensson-Egnund K (1982) Comparison of solid-phase immune electron microscopy, direct electron microscopy and enzyme-linked immunosorbent assay for detection of rotaviruses in faecal samples. J Virol Methods 4:45–53.
Kjeldsberg E, Siebke JC (1985) Use of immunosorbent electron microscopy for detection of rota-and hepatitis A virus in sucrose solutions. J Virol Methods 12:161–167.
Kleczkowski A (1961) Serological behaviour of tobacco mosaic virus and its protein fragments. Immunol. 4:130–141.
Lafferty KJ, Oertelis SJ (1961) Attachment of antibody to influenza virus. Nature 192:764–765.
Lafferty KJ, Oertelis S (1963) The interaction between virus and antibody. III. Examination of virus-antibody complexes with the electron microscope. Virology 21:91–99.
Lamontagne L, Marsolais G, Marois P, Assaf R (1980) Diagnosis of rotavirus, adenovirus, and herpesvirus infections by immune electron microscopy using a serum-in-agar diffusion method. Can J Microbiol 26:261–264.
Lee FK (1977) A study of enterovirus-antibody interaction in immunoelectron microscopy. M.Sc thesis, University of Toronto.
Lin N (1984) Gold IgG complexes improve the detection and identification of viruses in leaf dip preparations. J Virol Methods 8:181–190.
Milne RG, Luisoni E (1977) Rapid immune electron microscopy of virus preparations. In: Maramorosch K, Kaprowski H (eds) Methods in Virology. Academic Press, New York, vol 6, pp 265–281.
Morinet F, Ferchal F, Colimon R, Pérol Y (1984) Comparison of six methods for detecting human rotavirus in stools. Eur J Clin Microbiol 3:136–140.
Narang HK, Codd AA (1981) Frequency of pre-clumped virus in routine fecal specimens from patients with acute nonbacterial gastroenteritis. J Clin Microbiol 13:982–988.
Nicolaieff A, Obert G, Van Regenmortel MHV (1980) Detection of rotavirus by serological trapping on antibody-coated electron microscope grids. J Clin Microbiol 12:101–104.
Obert G, Gloekler R, Burckard J, Van Regenmortel MHV (1981) Comparison of immunosorbent electron microscopy, enzyme immunoassay and counter-immunoelectrophoresis for detection of human rotavirus in stools. J Virol Methods 3:99–107.
Pares RD, Whitecross MI (1982) Gold-labelled antibody decoration (GLAD) in the diagnosis of plant viruses by immune-electron microscopy. J Immunol Methods 51:23–28.
Paver W, Ashley C, Caul E, Clarke S (1973) A small virus in human faeces. Lancet 1:237–240.
Pegg-Feige (1983) Solid phase immunoelectron microscopy (SPIEM) in virus identification. M.Sc thesis. University of Toronto.
Pegg-Feige K, Doane FW (1983) Effect of specimen support film in solid phase immunoelectron microscopy. J Virol Methods 7:315–319.
Pegg-Feige K, Doane FW (1984) Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses. Proc of the 42nd Annual Meeting of the Electron Microscopy Society of America, pp 226-227.
Petrovicova A, Juck AS (1977) Serotyping of coxsackieviruses by immune electron microscopy. Acta Virol 21:165–167.
Rubenstein AS, Miller MF (1982) Comparison of an enzyme immunoassay with electron microscopic procedures for detecting rotavirus. J Clin Microbiol 15:938–944.
Stannard LM, Lennon M, Hodgkiss M, Smuts H (1982) An electron microscopic demonstration of immune complexes of hepatitis B e-antigen using colloidal gold as a marker. J Med Virol 9:165–175.
Svensson L, Grandien M, Pettersson C-A (1983) Comparison of solid-phase immune electron microscopy by use of protein A with direct electron microscopy and enzyme-linked immunosorbent assay for detection of rotavirus in stool. J Clin Microbiol 18:1244–1249.
van Rij G, Klepper L, Peperkamp E, Schaap GJP (1982) Immune electron microscopy and a cultural test in the diagnosis of adenovirus ocular infection. Brit J Ophthamol 66:317–319.
von Ardenne M., Friedrich-Freska H, Schramm G (1941) Elektronmikroskopische Untersuchung der pracipitinreaktion von tabakmosaikvirus mit kaninchenantiserum. Arch Gesamte Virusforsch 2:80–86.
Vreeswijk J, Folkers E, Wagenaar F, Kapsenberg JG (1988) The use of colloidal gold immunoelectron microscopy to diagnose varicella-zoster virus (VZV) infections by rapid discrimination between VZV, HSV-1 and HSV-2. J Virol Methods 22:255–271.
Watson DH, Wildy P (1963) Some serological properties of herpes virus particles studied with the electron microscope. Virology 21:100–111.
Wood DJ, Bailey AS (1987) Detection of adenovirus types 40 and 41 in stool specimens by immune electron microscopy. J Med Virol 21:191–199.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1991 Springer Science+Business Media New York
About this chapter
Cite this chapter
Doane, F.W., Anderson, N., Lee, F., Pegg-Feige, K., Hopley, J. (1991). Evaluation of Immunoassays for Electron Microscopy. In: de la Maza, L.M., Peterson, E.M. (eds) Medical Virology 9. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5856-5_1
Download citation
DOI: https://doi.org/10.1007/978-1-4684-5856-5_1
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-5858-9
Online ISBN: 978-1-4684-5856-5
eBook Packages: Springer Book Archive