Skip to main content
PMC home page
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 2010 Mar 25;55(1):88–101. doi: 10.1007/s12223-010-0015-8

The role of electron microscopy in the rapid diagnosis of viral infections — review

J Schramlová 1,✉, S Arientová 1, D Hulínská 1
PMCID: PMC7090635  PMID: 20336511

Abstract

Electron microscopy (EM) allows fast visualization of viruses in a wide range of clinical specimens. Viruses are grouped into families based on their morphology. Viruses from various families look distinctly and these morphological variances are the basis for identification of viruses by EM. The identification to the family level is often sufficient for the clinician or recognition of an unknown infectious agent. Diagnostic EM has two advantages over enzyme-linked immunosorbent assay and nucleic acid amplification tests. After a simple and fast negative staining, EM allows fast morphological identification and differential diagnosis of infectious agents contained in the specimen without the need for special considerations and/or reagents. Nevertheless, EM has the disadvantage of being unsuitable as a screening method.

Keywords: Gastroenteritis, Negative Staining, Rapid Diagnosis, Severe Acute Respiratory Syndrome, Nucleic Acid Amplification Test

Abbreviations

CSF

cerebrospinal fluid

ELISA

enzyme-linked immunosorbent assay

EM

electron microscopy

IEM

immune electron microscopy

NAT

nucleic acid amplification (techniques)

PBS

phosphate-buffered saline

PCR

polymerase chain reaction

PTA

phosphotungstic acid

SARS

severe acute respiratory syndrome

SPIEM

solid phase immune electron microscopy

NRL/ELM

National Reference Laboratory For Direct Diagnosis Of Viruses, Borreliae And Foreign Cell In Clinical Specimens

References

  1. Almeida J.D. Uses and abuses of diagnostic electron microscopy. Curr.Top.Microbiol.Immunol. 1983;104:147–157. doi: 10.1007/978-3-642-68949-9_9. [DOI] [PubMed] [Google Scholar]
  2. Appleton H.: Electron microscopy in virology, in J. Croker, D. Burnett (Eds): The Science of Laboratory Diagnosis, 2nd ed. Wiley 2005.
  3. Appleton H., Higgins P.G. Viruses and gastroenteritis in infants. Lancet. 1975;i:1297. doi: 10.1016/S0140-6736(75)92581-7. [DOI] [PubMed] [Google Scholar]
  4. Beards G.M. Laboratory diagnosis of viral gastroenteritis. Eur.J.Clin.Microbiol.Infect.Dis. 1988;7:11–13. doi: 10.1007/BF01962164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Biel S., Gelderblom H.R. Diagnostic electron microscopy is still a timely and rewarding method. J.Clin.Virol. 1999;3:105–119. doi: 10.1016/S1386-6532(99)00027-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Biel S.S., Nitsche A., Kurth A., Siegert W., Özel M., Gelderblom H.R. Detection of human polyomaviruses in urine from bone marrow transplant patients: a comparison of electron microscopy with PCR. Clin.Chem. 2004;50:306–312. doi: 10.1373/clinchem.2003.024539. [DOI] [PubMed] [Google Scholar]
  7. Brenner S., Horne R.W. A negative staining method for high resolution electron microscopy of viruses. Biochim.Biophys.Acta. 1959;34:103–110. doi: 10.1016/0006-3002(59)90237-9. [DOI] [PubMed] [Google Scholar]
  8. Clark B., Mckendrick M. A review of viral gastroenteritis. Curr.Opin.Infect.Dis. 2004;17:461–469. doi: 10.1097/00001432-200410000-00011. [DOI] [PubMed] [Google Scholar]
  9. Cotterill H.A., Macaulay M.E., Wong V. Reactivation of polyomavirus in bone marrow transplant recipients. J.Clin.Pathol. 1992;45:445. doi: 10.1136/jcp.45.5.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Curry A. Electron microscopy and the investigation of new infectious diseases. Internat.J.Infect.Dis. 2003;7:251–258. doi: 10.1016/S1201-9712(03)90103-2. [DOI] [PubMed] [Google Scholar]
  11. Curry A., Appleton H., Dowsett B. Application of transmission electron microscopy to the clinical study of viral and bacterial infections: present and future. Micron. 2006;37:91–106. doi: 10.1016/j.micron.2005.10.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Drosten C., Guenther S., Preiser W., Van Der Werf S., Brodt H.R., Becker S., Rabenau H., Panning M., Kolesnikova L., Fouchier R.A., Berger A., Burguiere A.M., Cinatl J., Eickmann M., Escriou N., Grywna K., Kramme S., Manuguerra J.C., Muller S., Rickerts V., Sturmer M., Vieth S., Klenk K.D., Osterhaus A.D., Schmitz H., Doerr H.W. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N.Engl.J.Med. 2003;348:1967–1976. doi: 10.1056/NEJMoa030747. [DOI] [PubMed] [Google Scholar]
  13. Falsey A.R., Walsh E.E. Novel coronavirus and severe acute respiratory syndrome. Lancet. 2003;361:1312–1313. doi: 10.1016/S0140-6736(03)13084-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Flewett T.H., Bryden A.S., Davies H.A.: Virus particles in gastroenteritis. Lancetii, 1497 (1973). [DOI] [PubMed]
  15. Frankenhauser R.L., Monroe S.S., Noel J.S., Humphrey C.D., Bresee J.S., Parashar U.D., Ando T., Glass R.I. Epidemiologic and molecular trends of Norwalk-like viruses associated with outbreaks of gastroenteritis in the United States. J.Infect.Dis. 2002;186:1–7. doi: 10.1086/341085. [DOI] [PubMed] [Google Scholar]
  16. Gallimore C.I., Green J., Richards A.F., Cotterill H., Curry A., Brown D.W., Gray J.J. Methods for the detection and characterization of noroviruses associated with outbreaks of gastroenteritis: outbreaks occurring in the north-west of England during two norovirus seasons. J.Med.Virol. 2004;73:280–288. doi: 10.1002/jmv.20088. [DOI] [PubMed] [Google Scholar]
  17. Gary G.W., Jr., Hierholzer J.C., Black R.E. Characteristics of noncultivable adenoviruses associated with diarrhea in infants: a new subgroup of human adenoviruses. J.Clin.Microbiol. 1979;10:96–106. doi: 10.1128/jcm.10.1.96-103.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gelderblom H.R., Renz H., Özel M. Negative staining in diagnostic virology. Micron Microscop.Acta. 1991;22:435–447. doi: 10.1016/0739-6260(91)90061-4. [DOI] [Google Scholar]
  19. Gentile M., Gelderblom H.R. Rapid viral diagnosis: role of electron microscopy. New Microbiol. 2005;28:1–12. [PubMed] [Google Scholar]
  20. Goldsmith C.S., Tatti K.M., Ksiazek T.G., Rollin P.E., Comer J.A., Lee W.W., Rota P.A., Bankamp B., Bellini W.J., Zaki S.R. Ultrasctructural characterization of SARS coronavirus. Emerg.Infect.Dis. 2004;10:320–326. doi: 10.3201/eid1002.030913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harris J. R. Negative Staining and Cryo-Electron Microscopy. Oxford: Bios Scientific Publications; 1997. [Google Scholar]
  22. Hawranek T., Trischer M., Muss W.H., Jecel J., Nowotny N., Kolodziejek J., Emberger M., Schnaeppi H., Hitner H. Feline orthopoxvirus infection transmitted from cat to human. Am.Acad.Dermatol. 2003;49:513–518. doi: 10.1067/S0190-9622(03)00762-X. [DOI] [PubMed] [Google Scholar]
  23. Hayat M.A., Miller S.S.: Negative Staining. McGraw-Hill Publishing Comp. 1990.
  24. Hazelton P.R., Gelderblom H.R. Electron microscopy for rapid diagnosis of infectious agents in emergent situations. Emerg.Infect. Dis. 2003;9:294–303. doi: 10.3201/eid0903.020327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hermann L., Enbree J., Hazelton P., Wells B., Coombs K. Reovirus type 2 isolated from cerebrospinal fluid. Pediatr.Dis.Infect. Dis.J. 2004;23:373–375. doi: 10.1097/00006454-200404000-00026. [DOI] [PubMed] [Google Scholar]
  26. Horne R.W. Virus Structure. New York: Academic Press; 1974. pp. 1–52. [Google Scholar]
  27. Horne R.W., Wildy P. Symmetry in virus architecture. Virology. 1961;15:348–373. doi: 10.1016/0042-6822(61)90366-X. [DOI] [PubMed] [Google Scholar]
  28. Horne R.W., Wildy P. Virus structure revealed by negative staining. Adv.Virus Res. 1963;10:101–170. doi: 10.1016/S0065-3527(08)60698-3. [DOI] [PubMed] [Google Scholar]
  29. Horne R.W., Wildy P. An historical account of the development and applications of the negative staining technique to the electron microscopy of viruses. J.Microscopy. 1979;117:103–122. doi: 10.1111/j.1365-2818.1979.tb00234.x. [DOI] [PubMed] [Google Scholar]
  30. Johnsen C.K., Böttiger B., Blom J. Confirmation of electron microscopy results by direct testing of viruses adhered to grids using nucleic acid amplification techniques. J.Virol.Methods. 2006;134:92–98. doi: 10.1016/j.jviromet.2005.12.007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kapikian A.Z. The discovery of the 27-nm Norwalk virus: an historic perspective. J.Infect.Dis. 2000;181(Suppl.2):S295–S302. doi: 10.1086/315584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kogasaka R., Nakamura S., Chiba S., Sakuma Y., Tera-Shima H., Yokoyama T., Nakao T. The 33-to 39-nm virus like particles tentatively designated as Sapporo agent, associated with an outbreak of acute gastroenteritis. J.Med.Virol. 1981;8:187–193. doi: 10.1002/jmv.1890080305. [DOI] [PubMed] [Google Scholar]
  33. Ksiazek T.G., Erdman D., Goldsmith C.S., Zak I.S.R., Peret T., Emery S., Tong S., Urbani C., Comer J.A., Lim W., Rollin P.E., Dowell S.F., Ling A.E., Humphrey C.D., Shieh W.J., Guarner J., Paddock C.D., Rota P., Fields B., Derisi J., Yang J.Y., Cox N., Hughes J.M., Leduc J.W., Bellini W.J., Anderson L.J., SARS Working Group A novel coronavirus associated with a severe respiratory syndrome. N.Engl.J.Med. 2003;348:153–196. doi: 10.1056/NEJMoa030781. [DOI] [PubMed] [Google Scholar]
  34. Lee T.W., Megson B., Kurtz J.B. Enterovirus typing by immune electron microscopy. J.Med.Microbiol. 1996;44:151–153. doi: 10.1099/00222615-44-2-151. [DOI] [PubMed] [Google Scholar]
  35. Lewis D.C., Lightfoot N.F., Pether J.V.S. Solid-phase immune electron microscopy with human immunoglobulin M for serotyping of Norwalk-like viruses. J.Clin.Microbiol. 1988;26:938–942. doi: 10.1128/jcm.26.5.938-942.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Madeley C.R. Viruses in the stool. J.Clin.Pathol. 1979;32:1–10. doi: 10.1136/jcp.32.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Madeley C.R. Viruses associated with acute diarrheal disease. In: Zuckermann A.J., Banatvala J.E., Pattison J.R., editors. Principles and Practice of Clinical Virology. 3rd ed. Chichester: Wiley; 1995. pp. 189–227. [Google Scholar]
  38. Madeley C.R. Origins of electron microscopy and viral diagnosis. J.Clin.Pathol. 1997;50:454–456. doi: 10.1136/jcp.50.6.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Madeley C.R. Diagnosing smallpox in possible bioterrorist attack. Lancet. 2003;361:97–98. doi: 10.1016/S0140-6736(03)12241-6. [DOI] [PubMed] [Google Scholar]
  40. Madeley C.R., Cosgrove B.P. 28-nm particles in feces in infantile gastroenteritis. Lancet. 1975;ii:451–452. doi: 10.1016/S0140-6736(75)90858-2. [DOI] [PubMed] [Google Scholar]
  41. Madeley C.R., Cosgrove B.P. Caliciviruses in man. Lancet. 1976;i:199–200. doi: 10.1016/S0140-6736(76)91309-X. [DOI] [PubMed] [Google Scholar]
  42. Madeley C.R., Field A.M. Virus Morphology. 2nd ed. Edinburgh: Churchill Livingstone; 1988. [Google Scholar]
  43. Mayo M.A. A summary of taxonomic changes recently approved by ICTV. Arch.Virol. 2002;147:1655–1656. doi: 10.1007/s007050200039. [DOI] [PubMed] [Google Scholar]
  44. Nermut M.V. General principles of virus architecture. In: Nermut M.V., Steven A.C., editors. Animal Virus Structure Perspectives in Medical Virology, Vol. 3. Amsterdam: Elsevier; 1987. pp. 3–18. [Google Scholar]
  45. Parashar U.D., Hummelmen E.G., Bresee J.S., Miller M.A., Glass R.I. Global illness and deaths caused by rotavirus disease in children. Emerg.Infect.Dis. 2003;9:565–572. doi: 10.3201/eid0905.020562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rao B., Basu A., Wairagkar N.S., Gore M.M., Arankalle V.A., Thakare J.P., Jadi R.S., Rao K.A., Mishra A.C. A large outbreak of acute encephalitis with high fertility rate in children in Andhra Pradesh, India in 2003 associated with Chandipura virus. Lancet. 2003;364:869–874. doi: 10.1016/S0140-6736(04)16982-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Reed K.D., Melski J.W., Graham M.B., Regnery R.L., Sotir M.J., Wegner M.V., Kazmierczak J.J., James J., Stratman E.J., Li Y., Fairley J.A., Swain G.R., Olson V.A., Sargent E.K., Kehl S.C., Frace M.A., Kline R., Foldy S.L., Davis J.P., Damon I.K. The detection of moneypox in humans in the Western hemisphere. N.Engl.J. Med. 2004;350:342–350. doi: 10.1056/NEJMoa032299. [DOI] [PubMed] [Google Scholar]
  48. Rýc M., Čiampor F., Wagner M. Electron Microscopy and Immune Electron Microscopy in Virology. Prague: Avicenum; 1989. pp. 140–154. [Google Scholar]
  49. Schimmer B., Sprenger H.G., Wismans P.J., van Genderren P.J. Three patients with orf (ecthyma contagiosum) Ned.Tijdschr. Geneeskd. 2004;148:788–791. [PubMed] [Google Scholar]
  50. Suzuki H., Sakai T., Tanabe N., Okabe N. Peak rotavirus activity shifted from winter to early spring in Japan. Pediatr.Infect.Dis.J. 2005;24:257–260. doi: 10.1097/01.inf.0000154327.00232.4d. [DOI] [PubMed] [Google Scholar]
  51. Trinidade G.S., Fonseca F.G., Marques J.T., Nogueira M.L., Mendes L.C., Borges A.S., Peiro J.R., Picuto E.M., Bonjardim C.A., Ferreira P.C., Kroon E.G. Aracatuba virus: a vaccinia-like virus associated with infection in humans and cattle. Emerg. Infect.Dis. 2003;9:155–160. doi: 10.3201/eid0902.020244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Tyrrell D.A.J., Almeida J. Direct electron-microscopy of organ cultures for the detection and characterization of viruses. Arch. Virol. 1967;22:417–425. doi: 10.1007/BF01242962. [DOI] [PubMed] [Google Scholar]
  53. van Regenmortel M.H.V., Fauquet C.M., Bishop D.H.L., Carstens E.B., Estes M.K., Lemon S.M., Maniloff J., Mayo M.A., Mcgeoch D.J., Pringle C.R., Wickner R.B. Virus Taxonomy: Classification and Nomenclature of Viruses, 7th Report of the. San Diego: International Committee on Taxonomy of Viruses. Academic Press; 2000. [Google Scholar]
  54. Vipond I.B., Caul E.O., Hirst D., Carmen B., Curry A., Lopman B.A., Pead P., Pickett M.A., Lambden P.R., Clarke I.N. National epidemic of Lordsdale norovirus in the UK. J.Clin.Virol. 2004;30:243–247. doi: 10.1016/j.jcv.2003.11.011. [DOI] [PubMed] [Google Scholar]

Articles from Folia Microbiologica are provided here courtesy of Nature Publishing Group

RESOURCES