Skip to main content

Advertisement

SpringerLink
Log in

Recovery of dengue virus from urine samples by real-time RT-PCR

  • Article
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Recently, reverse transcription polymerase chain reaction (RT-PCR) for dengue virus (DENV) has been reported to test positive in urine samples for a longer time frame than in serum. We evaluated two RNA extraction procedures from urine and investigated the stability of DENV RNA in urine and serum up to 1 year at different storage temperatures. In addition, 24 urine samples collected from patients with a recent infection were tested with DENV real-time RT-PCR and compared to the RT-PCR results on serum. Five patients with an acute DENV infection were followed up for 6 months by RT-PCR on urine. The automated extraction method with the MagNA Pure LC 2.0 device had a higher yield of DENV RNA compared to the manual QIAGEN method, explained by the higher volume used in the former method. DENV RNA in both serum and urine was stable at room temperature up to 1 month and at 4 °C and −20 °C for at least 1 year. The detection rate by RT-PCR on urine was 50 % (4/8) until day 7, 100 % (6/6) between 1 and 3 weeks after symptom onset, and 25 % (2/8) thereafter. Generally, DENV RNA concentrations are higher in serum than in urine up till day 7, switching to lower concentrations in serum thereafter. Peak concentrations in urine are reached around day 10, and RNA becomes undetectable 3 to 4 weeks following disease onset. This diagnostic tool is of added value in clinical settings by extending the period during which DENV infections are diagnosed by RT-PCR.

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
Fig. 2

Similar content being viewed by others

References

  1. Mackenzie JS, Gubler DJ, Petersen LR (2004) Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses. Nat Med 10:S98–S109

    Article  CAS  PubMed  Google Scholar 

  2. European Centre for Disease Control and Prevention (ECDC). Dengue fever. Factsheet for health professionals. Available online at: http://www.ecdc.europa.eu/en/healthtopics/dengue_fever/factsheet-for-health-professionals/Pages/Factsheet_health_professionals.aspx. Accessed 07 April 2014

  3. World Health Organization (WHO). Dengue and severe dengue. Fact sheet N°117. Available online at: http://www.who.int/mediacentre/factsheets/fs117/en/. Accessed 09 April 2014

  4. Odolini S, Parola P, Gkrania-Klotsas E, Caumes E, Schlagenhauf P, López-Vélez R, Burchard GD, Santos-O’Connor F, Weld L, von Sonnenburg F, Field V, de Vries P, Jensenius M, Loutan L, Castelli F (2012) Travel-related imported infections in Europe, EuroTravNet 2009. Clin Microbiol Infect 18:468–474

    Article  CAS  PubMed  Google Scholar 

  5. Van Esbroeck M, Verschueren J, Cnops L (2011) Significant increase in imported dengue infections in 2010 and an overview of 10 years of dengue surveillance in Belgium. Trop Med Int Health 16:I.4–035

    Article  Google Scholar 

  6. Panning M, Grywna K, Van Esbroeck M, Emmerich P, Drosten C (2008) Chikungunya fever in travelers returning to Europe from the Indian Ocean Region, 2006. Emerg Infect Dis 14:416–422

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Bottieau E, Van Esbroeck M, Cnops L, Clerinx J, Van Gompel A (2009) Chikungunya infection confirmed in a Belgian traveller returning from Phuket (Thailand). Euro Surveill 14:19248

    PubMed  Google Scholar 

  8. Schlagenhauf P, Weld L, Goorhuis A, Gautret P, Weber R, von Sonnenburg F, Lopez-Vélez R, Jensenius M, Cramer JP, Field VK, Odolini S, Gkrania-Klotsas E, Chappuis F, Malvy D, van Genderen PJ, Mockenhaupt F, Jauréguiberry S, Smith C, Beeching NJ, Ursing J, Parola P, Grobusch MP; EuroTravNet (2015) Travel-associated infection presenting in Europe (2008–12): an analysis of EuroTravNet longitudinal, surveillance data, and evaluation of the effect of the pre-travel consultation. Lancet Infect Dis 15:55–64

    Article  PubMed  Google Scholar 

  9. European Centre for Disease Prevention and Control (ECDC) (2013) Annual epidemiological report 2012. Chapter 2.4: emerging and vector-borne diseases

  10. Gjenero-Margan I, Aleraj B, Krajcar D, Lesnikar V, Klobučar A, Pem-Novosel I, Kurečić-Filipović S, Komparak S, Martić R, Duričić S, Betica-Radić L, Okmadžić J, Vilibić-Čavlek T, Babić-Erceg A, Turković B, Avsić-Županc T, Radić I, Ljubić M, Sarac K, Benić N, Mlinarić-Galinović G (2011) Autochthonous dengue fever in Croatia, August–September 2010. Euro Surveill 16:19805

    PubMed  Google Scholar 

  11. Schmidt-Chanasit J, Haditsch M, Schöneberg I, Günther S, Stark K, Frank C (2010) Dengue virus infection in a traveller returning from Croatia to Germany. Euro Surveill 15:19677

    PubMed  Google Scholar 

  12. Gould EA, Gallian P, de Lamballerie X, Charrel RN (2010) First cases of autochthonous dengue fever and chikungunya fever in France: from bad dream to reality! Clin Microbiol Infect 16:1702–1704

    Article  CAS  PubMed  Google Scholar 

  13. Marchand E, Prat C, Jeannin C, Lafont E, Bergmann T, Flusin O, Rizzi J, Roux N, Busso V, Deniau J, Noel H, Vaillant V, Leparc-Goffart I, Six C, Paty MC (2013) Autochthonous case of dengue in France, October 2013. Euro Surveill 18:20661

    CAS  PubMed  Google Scholar 

  14. Alves MJ, Fernandes PL, Amaro F, Osório H, Luz T, Parreira P, Andrade G, Zé-Zé L, Zeller H (2013) Clinical presentation and laboratory findings for the first autochthonous cases of dengue fever in Madeira Island, Portugal, October 2012. Euro Surveill 18:20398

    PubMed  Google Scholar 

  15. Hirayama T, Mizuno Y, Takeshita N, Kotaki A, Tajima S, Omatsu T, Sano K, Kurane I, Takasaki T (2012) Detection of dengue virus genome in urine by real-time reverse transcriptase PCR: a laboratory diagnostic method useful after disappearance of the genome in serum. J Clin Microbiol 50:2047–2052

    Article  PubMed Central  PubMed  Google Scholar 

  16. Korhonen EM, Huhtamo E, Virtala AM, Kantele A, Vapalahti O (2014) Approach to non-invasive sampling in dengue diagnostics: exploring virus and NS1 antigen detection in saliva and urine of travelers with dengue. J Clin Virol 61:353–358

    Article  PubMed  Google Scholar 

  17. Mizuno Y, Kotaki A, Harada F, Tajima S, Kurane I, Takasaki T (2007) Confirmation of dengue virus infection by detection of dengue virus type 1 genome in urine and saliva but not in plasma. Trans R Soc Trop Med Hyg 101:738–739

    Article  CAS  PubMed  Google Scholar 

  18. Ma X, Zhen W, Yang P, Sun X, Nie W, Zhang L, Xu H, Hu K (2014) First confirmation of imported dengue virus serotype 2 complete genome in urine from a Chinese traveler returning from India. Virol J 11:56–60

    Article  PubMed Central  PubMed  Google Scholar 

  19. Poloni TR, Oliveira AS, Alfonso HL, Galvão LR, Amarilla AA, Poloni DF, Figueiredo LT, Aquino VH (2010) Detection of dengue virus in saliva and urine by real time RT-PCR. Virol J 7:22

    Article  PubMed Central  PubMed  Google Scholar 

  20. European Centre for Disease Prevention and Control (ECDC) (2014) Dengue outbreak in Madeira, Portugal, March 2013. ECDC, Stockholm

  21. Johnson BW, Russell BJ, Lanciotti RS (2005) Serotype-specific detection of dengue viruses in a fourplex real-time reverse transcriptase PCR assay. J Clin Microbiol 43:4977–4983

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Cnops L, Franco L, Van Meensel B, Van den Ende J, Paz Sanchez-Seco M, Van Esbroeck M (2014) Three cases of imported dengue virus infection from Madeira to Belgium, 2012. J Travel Med 21:344–348

    Article  PubMed  Google Scholar 

  23. Basilío-de-Oliveira CA, Aguiar GR, Baldanza MS, Barth OM, Eyer-Silva WA, Paes MV (2005) Pathologic study of a fatal case of dengue-3 virus infection in Rio de Janeiro, Brazil. Braz J Infect Dis 9:341–347

    Article  PubMed  Google Scholar 

  24. Jessie K, Fong MY, Devi S, Lam SK, Wong KT (2004) Localization of dengue virus in naturally infected human tissues, by immunohistochemistry and in situ hybridization. J Infect Dis 189:1411–1418

    Article  PubMed  Google Scholar 

  25. Barzon L, Pacenti M, Franchin E, Squarzon L, Sinigaglia A, Ulbert S, Cusinato R, Palù G (2014) Isolation of West Nile virus from urine samples of patients with acute infection. J Clin Microbiol 52:3411–3413

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

This study was sponsored by the Belgian Society for Clinical Biology. The national reference center of West Nile virus (WNV) (and other arboviruses) is partially supported by the Belgian Ministry of Social Affairs through a fund with the Health Insurance System.

Compliance with ethical standards

This study was approved by the Institutional Review Board of the Institute of Tropical Medicine (ITM) and the Ethical Committee of the University Hospital of Antwerp. The study was carried out according to the principles stated in the Declaration of Helsinki, all applicable regulations, and according to established international scientific standards. Written consent for additional urine sampling was obtained from the five patients that were followed up for 6 months.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Central Laboratory of Clinical Biology, Department of Clinical Sciences, Institute of Tropical Medicine (ITM), Kronenburgstraat 43/3, 2000, Antwerp, Belgium

    D. Van den Bossche, L. Cnops & M. Van Esbroeck

Authors
  1. D. Van den Bossche
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L. Cnops
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Van Esbroeck
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D. Van den Bossche.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Van den Bossche, D., Cnops, L. & Van Esbroeck, M. Recovery of dengue virus from urine samples by real-time RT-PCR. Eur J Clin Microbiol Infect Dis 34, 1361–1367 (2015). https://doi.org/10.1007/s10096-015-2359-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-015-2359-0

Keywords

Search

Navigation