Skip to main content

Advertisement

SpringerLink
Log in

Dengue virus identification by transmission electron microscopy and molecular methods in fatal dengue hemorrhagic fever

  • Case Report
  • Published:
Infection Aims and scope Submit manuscript

Abstract

Dengue virus is the most significant virus transmitted by arthropods worldwide and may cause a potentially fatal systemic disease named dengue hemorrhagic fever. In this work, dengue virus serotype 4 was detected in the tissues of one fatal dengue hemorrhagic fever case using electron immunomicroscopy and molecular methods. This is the first report of dengue virus polypeptides findings by electron immunomicroscopy in human samples. In addition, not-previously-documented virus-like particles visualized in spleen, hepatic, brain, and pulmonary tissues from a dengue case are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

References

  1. Guzman MG, Halstead SB, Artsob H, Buchy P, Farrar J, Gubler DJ, et al. Dengue: a continuing global threat. Nat Rev Microbiol. 2010;8:S7–16. doi:10.1038/nrmicro2460.

    Article  PubMed  CAS  Google Scholar 

  2. San Martín JL, Brathwaite O, Zambrano B, Solórzano JO, Bouckenooghe A, Dayan GH, et al. The epidemiology of dengue in the Americas over the last three decades: a worrisome reality. Am J Trop Med Hyg. 2010;82:128–35. doi:10.4269/ajtmh.2010.09-0346.

    Article  PubMed  Google Scholar 

  3. World Health Organization (WHO). Dengue: guidelines for diagnosis, treatment, prevention and control. WHO. 2009.

  4. Kouri GP, Guzmán MG, Bravo JR. Why dengue haemorrhagic fever in Cuba? 2. An integral analysis. Trans R Soc Trop Med Hyg. 1987;81:821–3.

    Article  PubMed  CAS  Google Scholar 

  5. Guzmán MG, Kouri G, Valdes L, Bravo J, Alvarez M, Vazques S, et al. Epidemiologic studies on Dengue in Santiago de Cuba, 1997. Am J Epidemiol. 2000;152:793–9.

    Article  PubMed  Google Scholar 

  6. Guzmán MG, Kourí G. Dengue: an update. Lancet Infect Dis. 2002;2:33–42.

    Article  PubMed  Google Scholar 

  7. Peláez O, Guzmán MG, Kourí G, Pérez R, San Martín JL, Vázquez S, et al. Dengue 3 epidemic, Havana, 2001. Emerg Infect Dis. 2004;10:719–22.

    Article  PubMed  Google Scholar 

  8. OPS-Cuba. Brote de dengue en Cuba, 2006. EER Noticias: Enfermedades infecciosas emergentes y reemergentes, Región de Las Américas. 2006;3.

  9. García G, González N, Pérez AB, Sierra B, Aguirre E, Rizo D, et al. Long-term persistence of clinical symptoms in dengue-infected persons and its association with immunological disorders. Int J Infect Dis. 2011;15:e38–43.

    Article  PubMed  Google Scholar 

  10. Guzmán MG. Treinta años después de la epidemia cubana de dengue hemorrágico en 1981. Rev Cub Med Trop. 2012;64:5–14.

    Google Scholar 

  11. Pelegrino JL, Arteaga E, Rodríguez AJ, González E, Frontela MD, Guzmán MG. Normalizacion de tecnicas inmunohistoquimicas para la detección de antigenos del virus dengue en tejidos embebidos en parafina. Rev Cub Med Trop. 1997;49:100–7.

    CAS  Google Scholar 

  12. Limonta D, Capó V, Torres G, Pérez AB, Guzmán MG. Apoptosis in tissues from fatal dengue shock syndrome. J Clin Virol. 2007;40:50–4.

    Article  PubMed  CAS  Google Scholar 

  13. Rodriguez-Roche R, Alvarez M, Guzmán MG, Morier L, Kourí G. Comparison of rapid centrifugation assay with conventional tissue culture method for isolation of dengue 2 virus in C6/36-HT cells. J Clin Microbiol. 2000;38:3508–10.

    Google Scholar 

  14. Lanciotti RS, Calisher CH, Gubler DJ, Chang GJ, Vorndam AV. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol. 1992;30:545–51.

    PubMed  CAS  Google Scholar 

  15. Rosario D, Álvarez M, Vázquez S, Amin N, Rodríguez-Roche R, Guzmán MG. Application of molecular methods to the diagnosis and characterization of a dengue outbreak in Cuba. Biotecnologia Aplicada. 2001;18:203–6.

    CAS  Google Scholar 

  16. Callahan JD, Wu SJ, Dion-Schultz A, Mangold BE, Peruski LF, Watts DM, et al. Development and evaluation of serotype- and group-specific fluorogenic reverse transcriptase PCR (TaqMan) assays for dengue virus. J Clin Microbiol. 2001;39:4119–24.

    Article  PubMed  CAS  Google Scholar 

  17. Faulk WP, Taylor GM. An immunocolloid method for the electron microscope. Immunochemistry. 1971;8:1081–3.

    Article  PubMed  CAS  Google Scholar 

  18. Kellenberger E, Dürrenberger M, Villiger W, Carlemalm E, Wurtz M. The efficiency of immunolabel on Lowicryl sections compared to theoretical predictions. J Histochem Cytochem. 1987;35:959–69.

    Article  PubMed  CAS  Google Scholar 

  19. Hermida Díaz C, Pupo M, Guzmán Tirado MG, González Garriga M, Marcet Sánchez R. Empleo de un anticuerpo monoclonal anticomplejo dengue en la purificacion viral. Rev Cub Med Trop. 1992;44:171–6.

    Google Scholar 

  20. Fernández de Cossío ME, Díaz T, Galván A, Valdéz R, González E, Ayala M et al. Antigen recognition characteristics and comparative performance in immunoaffinity purification of two monoclonal antibodies specific for the hepatitis B virus surface antigen. J Biotechnol. 1997;56:69–80.

    Article  PubMed  Google Scholar 

  21. Guzmán MG, Alvarez M, Rodríguez R, Rosario D, Vázquez S, Valdés L, et al. Fatal dengue hemorrhagic fever in Cuba, 1997. Int J Infect Dis. 1999;3:130–5.

    Article  PubMed  Google Scholar 

  22. Pan American Health Organization (PAHO). Dengue and dengue hemorrhagic fever in the Americas: guidelines for prevention and control. Washington: PAHO. 1994.

  23. Bhamarapravati N, Tuchinda P, Boonyapaknavik V. Pathology of Thailand haemorrhagic fever: a study of 100 autopsy cases. Ann Trop Med Parasitol. 1967;61:500–10.

    PubMed  CAS  Google Scholar 

  24. Srikiatkhachorn A, Krautrachue A, Ratanaprakarn W, Wongtapradit L, Nithipanya N, Kalayanarooj S, et al. Natural history of plasma leakage in dengue hemorrhagic fever: a serial ultrasonographic study. Pediatr Infect Dis J. 2007;26:283–90.

    Article  PubMed  Google Scholar 

  25. Schmidt-Chanasit J, Tenner-Racz K, Poppert D, Emmerich P, Frank C, Dinges C, et al. Fatal dengue hemorrhagic fever imported into Germany. Infection. 2011. doi:10.1007/s15010-011-0208-3.

  26. Limonta D, González D, Capó V, Torres G, Pérez AB, Rosario D, et al. Fatal severe dengue and cell death in sickle cell disease during the 2001–2002 Havana dengue epidemic. Int J Infect Dis. 2009;13:e77–8.

    Article  PubMed  Google Scholar 

  27. de Araújo JM, Schatzmayr HG, de Filippis AM, Dos Santos FB, Cardoso MA, Britto C, et al. A retrospective survey of dengue virus infection in fatal cases from an epidemic in Brazil. J Virol Methods. 2009;155:34–8.

    Article  PubMed  Google Scholar 

  28. Boonpucknavig V, Bhamarapravati N, Boonpucknavig S, Futrakul P, Tanpaichitr P. Glomerular changes in dengue hemorrhagic fever. Arch Pathol Lab Med. 1976;100:206–12.

    PubMed  CAS  Google Scholar 

  29. Allwinn R, Schieferstein C, Glauke S, Doerr HW. Rapid diagnosis of primary dengue fever by the immunochromatographic test and by electron microscopy—a case report. Infection. 1999;27:365–7.

    Article  PubMed  CAS  Google Scholar 

  30. Noisakran S, Gibbons RV, Songprakhon P, Jairungsri A, Ajariyakhajorn C, Nisalak A, et al. Detection of dengue virus in platelets isolated from dengue patients. Southeast Asian J Trop Med Public Health. 2009;40:253–62.

    PubMed  CAS  Google Scholar 

  31. Wiersinga WJ, Scheepstra CG, Kasanardjo JS, de Vries PJ, Zaaijer H, Geerlings SE. Dengue fever-induced hemolytic uremic syndrome. Clin Infect Dis. 2006;43:800–1.

    Article  PubMed  Google Scholar 

  32. Lima Mda R, Nogueira RM, Schatzmayr HG, de Filippis AM, Limonta D, Dos Santos FB. A new approach to dengue fatal cases diagnosis: NS1 antigen capture in tissues. PLoS Negl Trop Dis. 2011;5:e1147. doi:10.1371/journal.pntd.0001147.

    Article  PubMed  Google Scholar 

  33. Hazelton PR, Gelderblom HR. Electron microscopy for rapid diagnosis of infectious agents in emergent situations. Emerg Infect Dis. 2003;9:294–303.

    Article  PubMed  Google Scholar 

  34. Goldsmith CS, Miller SE. Modern uses of electron microscopy for detection of viruses. Clin Microbiol Rev. 2009;22:552–63. doi:10.1128/CMR.00027-09.

    Article  PubMed  Google Scholar 

  35. Kulesh DA, Loveless BM, Norwood D, Garrison J, Whitehouse CA, Hartmann C, et al. Monkeypox virus detection in rodents using real-time 3′-minor groove binder TaqMan assays on the Roche LightCycler. Lab Invest. 2004;84:1200–8. doi:10.1038/labinvest.3700143.

    Article  PubMed  CAS  Google Scholar 

  36. Jessie K, Fong MY, Devi S, Lam SK, Wong KT. Localization of dengue virus in naturally infected human tissues, by immunohistochemistry and in situ hybridization. J Infect Dis. 2004;189:1411–8.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We are very grateful for the technical assistance and helpful comments during the manuscript preparation of Prof. Maritza Pupo from the Virology Department at the Pedro Kourí Tropical Medicine Institute (IPK). We acknowledge the staff of the Pathology Department and Intensive Care Unit of the Salvador Allende General Hospital in Havana. We are also grateful of Gilda Lemus from the Diagnostics Department of the Center for Biological Studies (CIB) in Havana for providing the anti-NS1 polyclonal antibodies.

Conflict of interest

The authors declare that there is no conflict of interest.

Ethical standards

This study received approval from the Ethics Committee of Pedro Kourí Tropical Medicine Institute (IPK).

Author information

Authors and Affiliations

  1. Arbovirus Laboratory, Virology Department, PAHO/WHO Collaborating Center for the Study of Dengue and its Vector, Pedro Kourí Tropical Medicine Institute (IPK), P.O. Box 601, Marianao 13, Havana, Cuba

    D. Limonta, D. Rosario, Y. Castellanos, M. Alvarez, R. Rodríguez-Roche, A. Pavón, J. Diaz & M. G. Guzmán

  2. Electron Microscopy Laboratory, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba

    V. Falcón, I. Menéndez & M. C. de la Rosa

  3. Pathology Department, Pedro Kourí Tropical Medicine Institute (IPK), Havana, Cuba

    G. Torres, V. Capó & L. López

  4. Pathology Department, Salvador Allende General Hospital, Havana, Cuba

    K. González

  5. Biomedical Research Division, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba

    G. Guillén

Authors
  1. D. Limonta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. V. Falcón
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Torres
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. V. Capó
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. I. Menéndez
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D. Rosario
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Y. Castellanos
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. M. Alvarez
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. R. Rodríguez-Roche
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. M. C. de la Rosa
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. A. Pavón
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. L. López
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. K. González
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. G. Guillén
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. J. Diaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. M. G. Guzmán
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. G. Guzmán.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Limonta, D., Falcón, V., Torres, G. et al. Dengue virus identification by transmission electron microscopy and molecular methods in fatal dengue hemorrhagic fever. Infection 40, 689–694 (2012). https://doi.org/10.1007/s15010-012-0260-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s15010-012-0260-7

Keywords

Search

Navigation