Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1983 Nov;18(5):1244–1249. doi: 10.1128/jcm.18.5.1244-1249.1983

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.

L Svensson, M Grandien, C A Pettersson
PMCID: PMC272873  PMID: 6315772

Abstract

A total of 525 stool specimens collected during 1 year were examined for the presence of rotavirus by direct electron microscopy (EM), enzyme-linked immunosorbent assay (ELISA), and a solid-phase immune electron microscope method (SPIEM) utilizing protein A-coated grids for anchoring of specific viral antisera. Rotavirus was seen in 187 specimens; SPIEM detected 183 (97.8%), whereas direct EM and ELISA detected 161 (86%) and 166 (88.7%), respectively. No false-positive reactions were seen by ELISA. The sensitivity of the methods was evaluated by coded investigation of a dilution series of a positive sample, with a negative fecal specimen as diluent. SPIEM was approximately 30 times more sensitive than direct EM and 10 times more sensitive than ELISA. A study was done to compare the elapsed time for recognition of rotavirus by SPIEM and EM in 25 randomly selected positive specimens. All virus-positive specimens were detected within 2 min by SPIEM, whereas up to 9 min was required for direct EM. SPIEM with protein A is a highly sensitive method, useful for rapid detection of viruses in clinical specimens. Due to the direct visualization of virus particles by electron microscopy, there is no requirement for monospecific antisera for the method.

Full text

PDF
1244

Images in this article

1246Image
on p.1246

Selected References

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

  1. Bishop R. F., Davidson G. P., Holmes I. H., Ruck B. J. Detection of a new virus by electron microscopy of faecal extracts from children with acute gastroenteritis. Lancet. 1974 Feb 2;1(7849):149–151. doi: 10.1016/s0140-6736(74)92440-4. [DOI] [PubMed] [Google Scholar]
  2. Brandt C. D., Kim H. W., Rodriguez W. J., Thomas L., Yolken R. H., Arrobio J. O., Kapikian A. Z., Parrott R. H., Chanock R. M. Comparison of direct electron microscopy, immune electron microscopy, and rotavirus enzyme-linked immunosorbent assay for detection of gastroenteritis viruses in children. J Clin Microbiol. 1981 May;13(5):976–981. doi: 10.1128/jcm.13.5.976-981.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Derrick K. S. Quantitative assay for plant viruses using serologically specific electron microscopy. Virology. 1973 Dec;56(2):652–653. doi: 10.1016/0042-6822(73)90068-8. [DOI] [PubMed] [Google Scholar]
  4. Flewett T. H., Bryden A. S., Davies H. Letter: Virus particles in gastroenteritis. Lancet. 1973 Dec 29;2(7844):1497–1497. doi: 10.1016/s0140-6736(73)92760-8. [DOI] [PubMed] [Google Scholar]
  5. Grauballe P. C., Genner J., Meyling A., Hornsleth A. Rapid diagnosis of rotavirus infections: comparison of electron microscopy and immunoelectroosmophoresis for the detection of rotavirus in human infantile gastroenteritis. J Gen Virol. 1977 May;35(2):203–218. doi: 10.1099/0022-1317-35-2-203. [DOI] [PubMed] [Google Scholar]
  6. Kapikian A. Z., Kim H. W., Wyatt R. G., Rodriguez W. J., Ross S., Cline W. L., Parrott R. H., Chanock R. M. Reoviruslike agent in stools: association with infantile diarrhea and development of serologic tests. Science. 1974 Sep 20;185(4156):1049–1053. doi: 10.1126/science.185.4156.1049. [DOI] [PubMed] [Google Scholar]
  7. Middleton P. J., Holdaway M. D., Petric M., Szymanski M. T., Tam J. S. Solid-phase radioimmunoassay for the detection of rotavirus. Infect Immun. 1977 May;16(2):439–444. doi: 10.1128/iai.16.2.439-444.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Middleton P. J., Petric M., Hewitt C. M., Szymanski M. T., Tam J. S. Counter-immunoelectro-osmophoresis for the detection of infantile gastroenteritis virus (orbi-group) antigen and antibody. J Clin Pathol. 1976 Mar;29(3):191–197. doi: 10.1136/jcp.29.3.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Narang H. K., Codd A. A. Frequency of preclumped virus in routine fecal specimens from patients with acute nonbacterial gastroenteritis. J Clin Microbiol. 1981 May;13(5):982–988. doi: 10.1128/jcm.13.5.982-988.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nicolaieff A., Obert G., van Regenmortel M. H. Detection of rotavirus by serological trapping on antibody-coated electron microscope grids. J Clin Microbiol. 1980 Jul;12(1):101–104. doi: 10.1128/jcm.12.1.101-104.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Obert G., Gloeckler R., Burckard J., van Regenmortel M. H. Comparison of immunosorbent electron microscopy, enzyme immunoassay and counterimmunoelectrophoresis for detection of human rotavirus in stools. J Virol Methods. 1981 Sep;3(2):99–107. doi: 10.1016/0166-0934(81)90006-9. [DOI] [PubMed] [Google Scholar]
  12. Rubenstein A. S., Miller M. F. Comparison of an enzyme immunoassay with electron microscopic procedures for detecting rotavirus. J Clin Microbiol. 1982 May;15(5):938–944. doi: 10.1128/jcm.15.5.938-944.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sanekata T., Yoshida Y., Okada H. Detection of rotavirus in faeces by latex agglutination. J Immunol Methods. 1981;41(3):377–385. doi: 10.1016/0022-1759(81)90199-x. [DOI] [PubMed] [Google Scholar]
  14. Sarkkinen H. K., Tuokko H., Halonen P. E. Comparison of enzyme-immunoassay and radioimmunoassay for detection of human rotaviruses and adenoviruses from stool specimens. J Virol Methods. 1980;1(6):331–341. doi: 10.1016/0166-0934(80)90050-6. [DOI] [PubMed] [Google Scholar]
  15. Spence L., Fauvel M., Petro R., Bloch S. Comparison of Counterimmunoelectrophoresis and electron microscopy for laboratory diagnosis of human reovirus-like agent-associated infantile gastroenteritis. J Clin Microbiol. 1977 Feb;5(2):248–249. doi: 10.1128/jcm.5.2.248-249.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Svensson L., von Bonsdorff C. H. Solid-phase immune electron microscopy (SPIEM) by use of protein A and its application for characterization of selected adenovirus serotypes. J Med Virol. 1982;10(4):243–253. doi: 10.1002/jmv.1890100404. [DOI] [PubMed] [Google Scholar]
  17. Yolken R. H., Kim H. W., Clem T., Wyatt R. G., Kalica A. R., Chanock R. M., Kapikian A. Z. Enzyme-linked immunosorbent assay (ELISA) for detection of human reovirus-like agent of infantile gastroenteritis. Lancet. 1977 Aug 6;2(8032):263–267. doi: 10.1016/s0140-6736(77)90951-5. [DOI] [PubMed] [Google Scholar]
  18. Yolken R. H., Stopa P. J. Analysis of nonspecific reactions in enzyme-linked immunosorbent assay testing for human rotavirus. J Clin Microbiol. 1979 Nov;10(5):703–707. doi: 10.1128/jcm.10.5.703-707.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zissis G., Lambert J. P. Enzyme-linked immunosorbent assays adapted for serotyping of human rotavirus strains. J Clin Microbiol. 1980 Jan;11(1):1–5. doi: 10.1128/jcm.11.1.1-5.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES