Archives of Virology

, Volume 71, Issue 3, pp 217–227 | Cite as

Virological diagnosis of enterovirus type 71 infections: Experiences gained during an epidemic of acute CNS diseases in Hungary in 1978

  • G. Nagy
  • Susanna Takátsy
  • Esther Kukán
  • Ilona Mihály
  • I. Dömök
Original Papers


In 1978 a severe epidemic of acute CNS diseases occurred in Hungary. Enterovirus type 71 (E71) proved to be the main aetiological agent. This could, however, be established only by special investigations, since the usual laboratory tests proved inefficient for diagnosis of E71 infections.

Attempts to isolate virus in tissue cultures were made on 1952 samples from 686 patients. E71 could only be isolated in cultures of a Vero cell line and even then only after 3 to 6 blind passages. Successful isolations were made from 47 stool samples, 5 throat swabs and 6 CNS samples originating from 44 patients. This represented only 13.6 per cent of the total number of E71 cases diagnosed in the course of studies, thus indicating the poor sensitivity of these tests. Suckling mice were unsuitable for the isolation of E71, although E71 tissue culture isolates proved pathogenic to these animals. Isolates could be identified by neutralization technique only after disaggregation of virions by ether treatment.

Blood samples from 1050 patients were tested for E71 antibodies by a radial plaque neutralization technique and those from 593 patients proved positive. A significant change in antibody level could only be detected in 52 out of 286 patients whose paired sera were available. The limited usefulness of this technique for the detection of current E71 infections was, however, indicated by the fact that 66 per cent of patients positive in virus isolation experiments already had antibodies in the early phase of their illness which remained at a constant level thereafter.

Investigation of 684 blood samples from 511 patients for E71-specific IgM antibodies showed this test to be the most reliable for the detection of current E71 infections. When appropriate blood samples were available, these antibodies could be detected in patients positive in the above mentioned tests, moreover, this method made an aetiological diagnosis possible in 236 cases which otherwise would have remained undetermined.

Infection with E71 was finally confirmed in 323 cases (poliomyelitis-like paralysis, 13; encephalitis, 145; aseptic meningitis, 161; hand, foot and mouth disease, 4).


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Baranyai, E., Káli, G.: Meningitis-meningoencephalitis epidemic among children caused by the enterovirus 71. (In Hungarian.) Orv. Hetil.121, 1175–1178 (1980).Google Scholar
  2. 2.
    Blomberg, J., Lycke, E., Ahlfors, K., Johnsson, T., Wolontis, S., von Zeipel, G.: New enterovirus type associated with epidemic of aseptic meningitis and/or hand, foot and mouth diseases. Lancet2, 112 (1974).Google Scholar
  3. 3.
    Chumakov, M. P., Lavrova, I. K., Martinova, L. I., Korolev, M. B., Bashkirtsev, V. N., Voroshilova, M. K.: Investigation of physicochemical properties of Bulgarian strain 258 of enterovirus type 71. Arch. Virol.60, 359–362 (1979).Google Scholar
  4. 4.
    Chumakov, M., Voroshilova, M., Shindarov, L., Lavrova, I., Grachova, L., Koroleva, G., Vasilenko, S., Brodvarova, I., Nikolova, M., Gyurova, S., Gacheva, M., Mitov, G., Ninov, N., Tsylka, E., Robinson, I., Frolova, M., Bashkirtsev, V., Martiyanova, L., Rodin, V.: Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria. Arch. Virol.60, 329–340 (1979).Google Scholar
  5. 5.
    Deibel, R., Gross, L.: Enterovirus type 71 infections. Observations in New York State in 1972–1977. In: Proc. 7th International Congress of Infectious and Parasitic Diseases, Varna, 1978, 366–370.Google Scholar
  6. 6.
    Deibel, R., Gross, L. L., Collins, D. N.: Isolation of a new enterovirus. Proc. Soc. Exp. Biol. (N.Y.)148, 203–207 (1975).Google Scholar
  7. 7.
    Dömök, I.: Isolation of Coxsackie virus strains from infections occurred in the years 1952–1954 and their identification. Acta microbiol. Acad. Sci. hung.3, 95–108 (1955).Google Scholar
  8. 8.
    Dömök, I., Molnár, E., Rudnai, O.: An epidemic of Bornholm disease in Hungary in 1958. Acta microbiol. Acad. Sci. hung.7, 151–168 (1960).Google Scholar
  9. 9.
    Dömök, I., Nagy, G., Takátsy, Zs., Kukán, E., György, E., Molnár, E., Simon, M., Hollós, I., Szarka, E.: Aetiological studies connected with the epidemic of acute CNS diseases occurred in Hungary in 1978. In preparation.Google Scholar
  10. 10.
    Gobara, F., Itagaki, A., Ito, Y., Saito, K., Katsumoto, T., Kimura, G.: Properties of virus isolated from an epidemic of hand-foot-and-mouth disease in 1973 in the city of Matsue. Microbiol. Immunol.21, 207–217 (1977).Google Scholar
  11. 11.
    Hagiwara, A., Tagaya, I., Yoneyama, T.: Epidemic of hand, foot and mouth disease associated with enterovirus 71 infection. Intervirology9, 60–63 (1978).Google Scholar
  12. 12.
    Kennet, M. L., Birch, C. J., Lewis, F. A., Yung, A. P., Locarnini, S. A., Gust, I. D.: Enterovirus Type 71 infection in Melbourne. Bull. World Hlth. Organ.51, 609–615 (1974).Google Scholar
  13. 13.
    Melnick, J. L., Schmidt, N. J., Mirkovic, R. R., Chumakov, M. P., Lavrova, I. K., Voroshilova, M. K.: Identification of Bulgarian strain 258 of enterovirus 71. Intervirol.12, 297–302 (1979).Google Scholar
  14. 14.
    Miwa, Ch., Ohtani, M., Watanabe, H., Fujimoto, J., Kinbara, K., Tanaka, H.: Epidemic of hand, foot and mouth disease in Gifu Prefecture in 1978. Jap. J. Med. Sci. Biol.33, 167–180 (1980).Google Scholar
  15. 15.
    Nagy, G., Mezey, I.: The use of ion exchange chromatography for demonstration of rubella-specific IgM antibodies. Acta microbiol. Acad. Sci. hung.24, 189–194 (1977).Google Scholar
  16. 16.
    Nagy, G., Mezey, I., Molnár, E.: Simple ion exchange batch technique for detection of IgM antibodies against rubella and flavivirus antigens. Acta microbiol. Acad. Sci. hung.24, 317–321 (1977).Google Scholar
  17. 17.
    Nagy, G., Takátsy, Zs.: Modified radial plaque neutralization test for demonstration of enterovirus neutralizing antibodies. Arch. Virol.65, 89–98 (1980).Google Scholar
  18. 18.
    Nikolova, M., Tsylka, E.: On the antigenic properties of strain 258 of enterovirus type 71 isolated in Bulgaria in 1975 (In Russian). Proc. 7th International Congress of Infectious and Parasitic Diseases, Varna, 1978, 392–395.Google Scholar
  19. 19.
    Ogra, P. L.: Distribution of echovirus antibody in serum, nasopharynx, rectum and spinal fluid after natural infection with echovirus type 6. Infect. Immun.2, 150–155 (1970).Google Scholar
  20. 20.
    Ogra, P. L., Karzon, D. T., Righthand, F., Mac Gillivray, M.: Immunoglobulin response in serum and secretions after immunization with live and inactivated poliovaccine and natural infection. New Engl. J. Med.279, 893–900 (1968).Google Scholar
  21. 21.
    Schmidt, N. J.: Tissue culture techniques for diagnostic virology. In:Lennette, E. H., Schmidt, N. J. (eds.): Diagnostic procedures for viral and rickettsial infections, 4th ed., 79–178. New York: American Public Health Association 1969.Google Scholar
  22. 22.
    Schmidt, N. J., Guenther, R. W., Lennette, E. H.: Typing of ECHO virus isolates by immune serum pools. The “intersecting serum scheme”. J. Immunol.87, 623–626 (1961).Google Scholar
  23. 23.
    Schmidt, N. J., Lennette, E. H., Dennis, J.: Characterization of antibodies produced in natural and experimental coxsackie-virus infections. J. Immunol.100, 99–106 (1968).Google Scholar
  24. 24.
    Schmidt, N. J., Lennette, E. H., Ho, H. H.: An apparently new enterovirus isolated from patients with disease of the central nervous system. J. Inf. Dis.129, 304–309 (1974).Google Scholar
  25. 25.
    Shindarov, L. M., Chumakov, M. P., Voroshilova, M. K., Bojinov, S., Vasilenko, S. M., Iordanov, I., Kirov, I. D., Kamenov, E., Leshchinskaya, E. V., Mitov, G., Robinson, I. A., Sivchev, S., Staikov, St.: Epidemiological, clinical, and pathomorphological characteristics of epidemic poliomyelitis-like disease caused by enterovirus 71. J. Hyg. Epid. Microbiol. Immunol.23, 284–295 (1979).Google Scholar
  26. 26.
    Solt, K.: In preparation.Google Scholar
  27. 27.
    Tariska, I., Mázló, M.: Neuropathology of encephalomyelitis caused by enterovirus 71 infection. (In Hungarian.) Orv. Hetil.121, 1739–1743 (1980).Google Scholar
  28. 28.
    von Zeipel, G.: Disaggregation in strains of enterovirus 71 and echovirus type 4 after propagation in human RD cells. In: Proc. 7th International Congress of Infectious and Parasitic Diseases, Varna, 1978, 389–391.Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • G. Nagy
    • 1
  • Susanna Takátsy
    • 1
  • Esther Kukán
    • 2
  • Ilona Mihály
    • 2
  • I. Dömök
    • 1
  1. 1.Department of VirologyNational Institute of HygieneBudapestHungary
  2. 2.Virus LaboratoryCentral Hospital for Infectious DiseasesBudapestHungary

Personalised recommendations