Volume 84, Issue 2
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Dengue viral isolation is necessary for definitive diagnosis, pathogenesis and evolutionary research, vaccine candidates, and diagnostic materials. Using standardized techniques, we analyzed isolation rates of 1,544 randomly selected polymerase chain reaction (PCR)-positive samples, representing all four dengue serotypes, from patients with serologically confirmed dengue infections and evaluated whether clinical and laboratory results could be predictive of isolation using standard and mosquito isolation techniques. Viruses were isolated from 62.5% of the samples by direct application to C6/36 cells and increased to 79.4% when amplifying C6/36 negative samples by intrathorasic inoculation in mosquitoes. High viremia, measured by reverse transcriptase (RT)-PCR, was a strong predictor for viral isolation by either method. Isolation was most successful in samples collected early in the disease, had low antibody levels, temperatures greater than 38°C, and had a final clinical diagnosis of dengue fever. Dengue serotypes also played a role in the success of viral isolation.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. Gibbons RV, Vaughn DW, , 2002. Dengue: an escalating problem. BMJ 324: 15631566.[Crossref] [Google Scholar]
  2. Mackenzie JS, Gubler DJ, Petersen LR, , 2004. Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses. Nat Med 10: S98S109.[Crossref] [Google Scholar]
  3. Anonymous, 1997. Dengue Hemorrhagic Fever: Diagnosis, Treatment, Prevention and Control. Second edition. Geneva, Switzerland: World Health Organization. [Google Scholar]
  4. Burke DS, Nisalak A, Johnson DE, Scott RM, , 1988. A prospective study of dengue infections in Bangkok. Am J Trop Med Hyg 38: 172180. [Google Scholar]
  5. Guzman MG, Kouri GP, Bravo J, Soler M, Vazquez S, Morier L, , 1990. Dengue hemorrhagic fever in Cuba, 1981: a retrospective seroepidemiologic study. Am J Trop Med Hyg 42: 179184. [Google Scholar]
  6. Halstead SB, Chow JS, Marchette NJ, , 1973. Immunological enhancement of dengue virus replication. Nat New Biol 243: 2426. [Google Scholar]
  7. Halstead SB, , 2002. Dengue hemorrhagic fever: two infections and antibody dependent enhancement, a brief history and personal memoir. Rev Cubana Med Trop 54: 171179. [Google Scholar]
  8. Johnson AJ, Martin DA, Karabatsos N, Roehrig JT, , 2000. Detection of anti-arboviral immunoglobulin G by using a monoclonal antibody-based capture enzyme-linked immunosorbent assay. J Clin Microbiol 38: 18271831. [Google Scholar]
  9. Scott RM, McCown JM, Russell PK, , 1972. Human immunoglobulin specificity after group B arbovirus infections. Infect Immun 6: 277281. [Google Scholar]
  10. Allwinn R, Doerr HW, Emmerich P, Schmitz H, Preiser W, , 2002. Cross-reactivity in flavivirus serology: new implications of an old finding? Med Microbiol Immunol (Berl) 190: 199202.[Crossref] [Google Scholar]
  11. Kalayanarooj S, Nimmannitya S, Suntayakorn S, Vaughn DW, Nisalak A, Green S, Chanisiriwongs V, Rothman A, Ennis FA, , 1999. Can doctors make an accurate diagnosis if dengue infections at an early stage? Dengue Bull 23: 19. [Google Scholar]
  12. Lanciotti RS, Calisher CH, Gubler DJ, Chang GJ, Vorndam AV, , 1992. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 30: 545551. [Google Scholar]
  13. Klungthong C, Gibbons RV, Thaisomboonsuk B, Nisalak A, Kalayanarooj S, Thirawuth V, Nutkamhang N, Mammen MP, Jarman RG, , 2007. Dengue virus detection using whole blood for RT-PCR and viral isolation. J Clin Microbiol 45: 24802485.[Crossref] [Google Scholar]
  14. Singh KR, Paul SD, , 1969. Isolation of dengue viruses in Aedes albopictus cell cultures. Bull World Health Organ 40: 982983. [Google Scholar]
  15. Tesh RB, , 1979. A method for the isolation and identification of dengue viruses, using mosquito cell cultures. Am J Trop Med Hyg 28: 10531059. [Google Scholar]
  16. Kuno G, Gomez I, Gubler DJ, , 1991. An ELISA procedure for the diagnosis of dengue infections. J Virol Methods 33: 101113.[Crossref] [Google Scholar]
  17. Rosen L, , 1981. The use of Toxorhynchites mosquitoes to detect and propagate dengue and other arboviruses. Am J Trop Med Hyg 30: 177183. [Google Scholar]
  18. Rosen L, Gubler DJ, , 1974. The use of mosquitoes to detect and propagate dengue viruses. Am J Trop Med Hyg 23: 11531160. [Google Scholar]
  19. Rosen L, Shroyer DA, , 1985. Comparative susceptibility of five species of Toxorhynchites mosquitoes to parenteral infection with dengue and other flaviviruses. Am J Trop Med Hyg 34: 805809. [Google Scholar]
  20. Watts DM, Harrison BA, Nisalak A, Scott RM, Burke DS, , 1982. Evaluation of Toxorhynchites splendens (Diptera:Culicidae) as a bioassay host for dengue viruses. J Med Entomol 19: 5459.[Crossref] [Google Scholar]
  21. Innis BL, Nisalak A, Nimmannitya S, Kusalerdchariya S, Chongswasdi V, Suntayakorn S, Puttisri P, Hoke CH, Jr, 1989. An enzyme-linked immunosorbent assay to characterize dengue infections where dengue and Japanese encephalitis co-circulate. Am J Trop Med Hyg 40: 418427. [Google Scholar]
  22. Blacksell SD, Bell D, Kelley J, Mammen MP, Jr Gibbons RV, Jarman RG, Vaughn DW, Jenjaroen K, Nisalak A, Thongpaseuth S, Vongsouvath M, Davong V, Phouminh P, Phetsouvanh R, Day NP, Newton PN, , 2007. Prospective study to determine accuracy of rapid serological assays for the diagnosis of acute dengue virus infection in Laos. Clin Vaccine Immunol 14: 14581464.[Crossref] [Google Scholar]
  23. Vaughn DW, Nisalak A, Kalayanarooj S, Solomon T, Kneen R, Dung NM, Cuzzubbo A, Devine PL, , 1998. Evaluation of rapid commercial assays for the diagnosis of dengue infection. Annual Meeting of the American Society of Microbiology . Atlanta, GA, May 17–21, 1997. [Google Scholar]
  24. Vaughn DW, Nisalak A, Kalayanarooj S, Solomon T, Dung NM, Cuzzubbo A, Devine PL, , 1998. Evaluation of a rapid immunochromatographic test for the diagnosis of dengue infection. J Clin Microbiol 36: 234238. [Google Scholar]
  25. Deubel V, Laille M, Hugnot JP, Chungue E, Guesdon JL, Drouet MT, Bassot S, Chevrier D, , 1990. Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood. J Virol Methods 30: 4154.[Crossref] [Google Scholar]
  26. Henchal EA, Polo SL, Vorndam V, Yaemsiri C, Innis BL, Hoke CH, Jr, 1991. Sensitivity and specificity of a universal primer set for the rapid diagnosis of dengue virus infections by polymerase chain reaction and nucleic acid hybridization. Am J Trop Med Hyg 45: 418428. [Google Scholar]
  27. Oliveira De Paula S, Malta Lima D, Clotteau M, Pires Neto Rd Rda J, Lopes da Fonseca BA, , 2003. Improved detection of dengue-1 virus from IgM-positive serum samples using C6/36 cell cultures in association with RT-PCR. Intervirology 46: 227231. [Google Scholar]
  28. Nisalak A, Endy TP, Nimmannitya S, Kalayanarooj S, Thisayakorn U, Scott RM, Burke DS, Hoke CH, Innis BL, Vaughn DW, , 2003. Serotype-specific dengue virus circulation and dengue disease in Bangkok, Thailand from 1973 to 1999. Am J Trop Med Hyg 68: 191202. [Google Scholar]
  29. Vaughn DW, Green S, Kalayanarooj S, Innis BL, Nimmannitya S, Suntayakorn S, Endy TP, Raengsakulrach B, Rothman AL, Ennis FA, Nisalak A, , 2000. Dengue viremia titer, antibody response pattern, and virus serotype correlate with disease severity. J Infect Dis 181: 29.[Crossref] [Google Scholar]
  30. Libraty DH, Endy TP, Houng HS, Green S, Kalayanarooj S, Suntayakorn S, Chansiriwongs W, Vaughn DW, Nisalak A, Ennis FA, Rothman AL, , 2002. Differing influences of virus burden and immune activation on disease severity in secondary dengue-3 virus infections. J Infect Dis 185: 12131221.[Crossref] [Google Scholar]
  31. Wang WK, Chao DY, Kao CL, Wu HC, Liu YC, Li CM, Lin SC, Ho ST, Huang JH, King CC, , 2003. High levels of plasma dengue viral load during defervescence in patients with dengue hemorrhagic fever: implications for pathogenesis. Virology 305: 330338.[Crossref] [Google Scholar]
  32. Wang WK, Chen HL, Yang CF, Hsieh SC, Juan CC, Chang SM, Yu CC, Lin LH, Huang JH, King CC, , 2006. Slower rates of clearance of viral load and virus-containing immune complexes in patients with dengue hemorrhagic fever. Clin Infect Dis 43: 10231030.[Crossref] [Google Scholar]
  33. Thomas L, Verlaeten O, Cabie A, Kaidomar S, Moravie V, Martial J, Najioullah F, Plumelle Y, Fonteau C, Dussart P, Cesaire R, , 2008. Influence of the dengue serotype, previous dengue infection, and plasma viral load on clinical presentation and outcome during a dengue-2 and dengue-4 co-epidemic. Am J Trop Med Hyg 78: 990998. [Google Scholar]
  34. Vaughn DW, Green S, Kalayanarooj S, Innis BL, Nimmannitya S, Suntayakorn S, Rothman AL, Ennis FA, Nisalak A, , 1997. Dengue in the early febrile phase: viremia and antibody responses. J Infect Dis 176: 322330.[Crossref] [Google Scholar]
  35. Yamada K, Takasaki T, Nawa M, Kurane I, , 2002. Virus isolation as one of the diagnostic methods for dengue virus infection. J Clin Virol 24: 203209.[Crossref] [Google Scholar]

Data & Media loading...

  • Received : 30 Dec 2009
  • Accepted : 26 Oct 2010
  • Published online : 04 Feb 2011

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error